AP1S1 missense mutations cause a congenital enteropathy via an epithelial barrier defect.

Congenital diarrheal disorders (CDD) comprise > 50 monogenic entities featuring chronic diarrhea of early-onset, including defects in nutrient and electrolyte absorption, enterocyte polarization, enteroendocrine cell differentiation, and epithelial integrity. Diarrhea is also a predominant symptom in many immunodeficiencies, congenital disorders of glycosylation, and in some defects of the vesicular sorting and transporting machinery. We set out to identify the etiology of an intractable diarrhea in 2 consanguineous families by whole-exome sequencing, and identified two novel AP1S1 mutations, c.269T>C (p.Leu90Pro) and c.346G>A (p.Glu116Lys). AP1S1 encodes the small subunit of the adaptor protein 1 complex (AP-1), which plays roles in clathrin coat-assembly and trafficking between trans-Golgi network, endosomes and the plasma membrane. An AP1S1 knock-out (KO) of a CaCo2 intestinal cell line was generated to characterize intestinal AP1S1 deficiency as well as identified mutations by stable expression in KO background. Morphology and prototype transporter protein distribution were comparable between parental and KO cells. We observed altered localization of tight-junction proteins ZO-1 and claudin 3, decreased transepithelial electrical resistance and an increased dextran permeability of the CaCo2-AP1S1-KO monolayer. In addition, lumen formation in 3D cultures of these cells was abnormal. Re-expression of wild-type AP1S1 in CaCo2-AP1S1-KO cells reverted these abnormalities, while expression of AP1S1 containing either missense mutation did not. Our data indicate that loss of AP1S1 function causes an intestinal epithelial barrier defect, and that AP1S1 mutations can cause a non-syndromic form of congenital diarrhea, whereas 2 reported truncating AP1S1 mutations caused MEDNIK syndrome, characterized by mental retardation, enteropathy, deafness, neuropathy, ichthyosis, and keratodermia.

A CRISPR screen in intestinal epithelial cells identifies novel factors for polarity and apical transport.

Epithelial polarization and polarized cargo transport are highly coordinated and interdependent processes. In our search for novel regulators of epithelial polarization and protein secretion, we used a genome-wide CRISPR/Cas9 screen and combined it with an assay based on fluorescence-activated cell sorting (FACS) to measure the secretion of the apical brush-border hydrolase dipeptidyl peptidase 4 (DPP4). In this way, we performed the first CRISPR screen to date in human polarized epithelial cells. Using high-resolution microscopy, we detected polarization defects and mislocalization of DPP4 to late endosomes/lysosomes after knockout of TM9SF4, anoctamin 8, and ARHGAP33, confirming the identification of novel factors for epithelial polarization and apical cargo secretion. Thus, we provide a powerful tool suitable for studying polarization and cargo secretion in epithelial cells. In addition, we provide a dataset that serves as a resource for the study of novel mechanisms for epithelial polarization and polarized transport and facilitates the investigation of novel congenital diseases associated with these processes.

Skin manifestations in rare types of diabetes and other endocrine conditions.

As the most visible and vulnerable organ of the human organism, the skin can provide an impression of its state of health. Rare forms of diabetes and endocrinopathies are often diagnosed late or primarily misinterpreted due to their rarity. Skin peculiarities associated with these rare diseases may be indicative of the underlying endocrinopathy or form of diabetes. At the same time, rare skin changes in diabetes or endocrinopathies can also be a major challenge for dermatologists, diabetologists and endocrinologists in optimal patient and therapy management. Active collaboration between these different specialist groups can therefore lead to increased patient safety, better therapeutic success and more targeted diagnostics.

Collaboration for rare diabetes: understanding new treatment options for Wolfram syndrome.

BACKGROUND: Wolfram Syndrome is a very rare genetic disease causing diabetes mellitus, blindness, deafness, diabetes insipidus, and progressive brainstem degeneration. Neurologic symptoms of affected patients include ataxia, sleep apnea, loss of bladder control, dysphagia, loss of taste, and accompanying psychiatric symptoms as a sign of progressive neurodegeneration. Its genetic cause is mainly biallelic mutations of the Wolframin endoplasmatic reticulum transmembrane glycoprotein gene Wfs1. These result in increased ER stress, which in turn induces apoptosis and leads to the depletion of the corresponding cells and a loss of their physiological functions. Though diabetes mellitus is mostly treated by insulin, there is still no proven cure for the disease in general. It leads to premature death in affected individuals-usually within the 4th decade of live. CURRENT RESEARCH AND TREATMENT TRIALS: Clinical studies are currently being conducted at various locations worldwide to test a therapy for the disease using various approaches. POTENTAIL OF VIRTUAL NETOWRKING: As rare diseases in general represent a major challenge for individual clinicians and researchers due to the rarity of diagnosis, the lack of evidence and of value of existing research, international cooperation, coordination and networking leading to an alignment of different stakeholders is necessary to support patients and increase knowledge about these diseases, like wolfram syndrome. CONCLUSION: ENDO-ERN and EURRECA are two EU-funded networks that aim to promote knowledge sharing, education and research on rare endocrine diseases.

Peri-interventional combined anticoagulation and antithrombotic therapy in atrial fibrillation ablation: A retrospective safety analysis.

BACKGROUND: Catheter ablation (CA) of atrial fibrillation (AF) requires an intensified peri-inter-ventional anticoagulation scheme to avoid thromboembolic complications. In patients with cardiac or extracardiac artery disease, an additional antiplatelet treatment (AAT) is at least temporally necessary especially after a percutaneous intervention with stent implantation. This raises the question whether these patients have a higher peri-interventional bleeding risk during CA of AF. METHODS: The data of 1235 patients with CA of AF were retrospectively analyzed in terms of bleeding events, ablation type, antithrombotic medication and comorbidities such as coronary artery disease and components of the HAS- BLED score. Peri-interventional bleeding events were classified in accordance with the BARC classification. Differentiations were made between slight femoral bleeding (based on type 1), severe femoral bleeding and pericardial effusion without pericardiocentesis (based on type 2) with the need of further hospitalization, the need of transfusion (based on type 3a) and pericardial tamponades requiring pericardiocentesis (based on type 3b). RESULTS: 1131/1235 (91.6%) patients were exclusively under anticoagulation and 187 (15.3%) patients were also on AAT. There were no statistically significant differences in type 1 and 3b bleeding complica-tions or the occurrence of femoral pseudoaneurysms between both groups. However, type 2/3a bleeding complications, mostly femoral bleedings, were significantly more frequent in the patient group with AAT (3.2% vs. 7.5%, p = 0.006). CONCLUSIONS: An additional antiplatelet therapy increases the risk of severe femoral bleeding events during CA of AF. It appears reasonable to perform the elective procedure of AF ablation after the dis-continuation of AAT.

Unilateral recurrent nerve palsy and cardiovascular disease - Ortner's syndrome.

Recurrent laryngeal palsies are relatively common. Frequently, surgical procedures precede paresis. In rare cases a compression of the recurrent laryngeal nerve can be caused by enlarged cardiovascular structures. The phenomenon of compression of the left recurrent laryngeal nerve is explicitly designated as Ortner's syndrome, first described in 1897 by Norbert Ortner. Nowadays the compression of the right recurrent laryngeal nerve by cardiovascular structures is also associated with Ortner's syndrome. We report two cases of an 82- and a 71-year-old patient who presented with hoarseness and each right- and left-sided vocal cord paralysis for further diagnosis. The ear, nose, and throat (ENT) examinations revealed no clarifying findings besides the vocal cord palsy, so extensive imaging techniques were used. The cause of left-sided recurrent laryngeal nerve palsy was a penetrated aortic ulcer caused by large thrombosed aneurysm of the aortic arch. The right-sided paresis arose due to aneurysmal enlargement of the brachiocephalic trunk and an aneurysm of right subclavian artery. These cases demonstrate that interdisciplinary medical work is important. The internal medical presentation of a patient with hoarseness without ENT medical findings should be considered. .

In Patients Undergoing Cochlear Implantation, Psychological Burden Affects Tinnitus and the Overall Outcome of Auditory Rehabilitation.

Cochlear implantation (CI) is increasingly being used in the auditory rehabilitation of deaf patients. Here, we investigated whether the auditory rehabilitation can be influenced by the psychological burden caused by mental conditions. Our sample included 47 patients who underwent implantation. All patients were monitored before and 6 months after CI. Auditory performance was assessed using the Oldenburg Inventory (OI) and Freiburg monosyllable (FB MS) speech discrimination test. The health-related quality of life was measured with Nijmegen Cochlear implantation Questionnaire (NCIQ) whereas tinnitus-related distress was measured with the German version of Tinnitus Questionnaire (TQ). We additionally assessed the general perceived quality of life, the perceived stress, coping abilities, anxiety levels and the depressive symptoms. Finally, a structured interview to detect mental conditions (CIDI) was performed before and after surgery. We found that CI led to an overall improvement in auditory performance as well as the anxiety and depression, quality of life, tinnitus distress and coping strategies. CIDI revealed that 81% of patients in our sample had affective, anxiety, and/or somatoform disorders before or after CI. The affective disorders included dysthymia and depression, while anxiety disorders included agoraphobias and unspecified phobias. We also diagnosed cases of somatoform pain disorders and unrecognizable figure somatoform disorders. We found a positive correlation between the auditory performance and the decrease of anxiety and depression, tinnitus-related distress and perceived stress. There was no association between the presence of a mental condition itself and the outcome of auditory rehabilitation. We conclude that the CI candidates exhibit high rates of psychological disorders, and there is a particularly strong association between somatoform disorders and tinnitus. The presence of mental disorders remained unaffected by CI but the degree of psychological burden decreased significantly post-CI. The implants benefitted patients in a number of psychosocial areas, improving the symptoms of depression and anxiety, tinnitus, and their quality of life and coping strategies. The prevalence of mental disorders in patients who are candidates for CI suggests the need for a comprehensive psychological and psychosomatic management of their treatment.

Deceleration capacity of heart rate predicts 1-year mortality of patients undergoing transcatheter aortic valve implantation.

BACKGROUND: Risk prediction in patients with severe aortic stenosis (AS) undergoing transcatheter aortic valve implantation (TAVI) is challenging. Development of novel markers for patient risk assessment is of great clinical value. Deceleration capacity (DC) of heart rate is a strong risk predictor in post-infarction patients. HYPOTHESIS: DC provides prognostic information in patients undergoing TAVI. METHODS: We enrolled 374 consecutive patients with severe AS undergoing TAVI. All patients received 24-hour Holter recording or continuous heart-rate monitoring to assess DC before intervention. Primary endpoint was all-cause mortality after 1 year. RESULTS: Forty-nine patients (13.1%) died within 1 year. DC was significantly lower in nonsurvivors than in survivors (1.2 ± 4.8 ms vs 3.3 ± 2.9 ms; P < 0.001), whereas the logistic EuroSCORE and EuroSCORE II were comparable between groups (logistic EuroSCORE: 27.3% ± 17.0% vs 22.9% ± 14.2%; P = 0.122; EuroSCORE II: 8.0% ± 6.9% vs 6.7% ± 4.8%, P = 0.673). One-year mortality in the 116 patients with impaired DC (<2.5 ms) was significantly higher than in patients with normal DC (23.3% vs 8.5%; P < 0.001). In multivariate Cox regression analysis that included DC, sex, paroxysmal atrial fibrillation, hemoglobin level before TAVI, and logistic EuroSCORE, DC was the strongest predictor of 1-year mortality (hazard ratio: 0.88, 95% confidence interval: 0.85-0.94, P < 0.001). DC yielded an AUC in the ROC analysis of 0.645. CONCLUSIONS: DC of heart rate is a strong and independent predictor of 1-year mortality in patients with severe AS undergoing TAVI.

Deceleration capacity as a risk predictor in patients presenting to the emergency department with syncope: A prospective exploratory pilot study.

Syncope is a common cause for admission to the emergency department (ED). Due to limited clinical resources there is great interest in developing risk stratification tools that allow identifying patients with syncope who are at low risk and can be safely discharged. Deceleration capacity (DC) is a strong risk predictor in postinfarction and heart failure patients. The aim of this study was to evaluate whether DC provides prognostic information in patients presenting to ED with syncope.We prospectively enrolled 395 patients presenting to the ED due to syncope. Patient's electrocardiogram (ECG) for the calculation of DC was recorded by monitoring devices which were started after admission. Both the modified early warning score (MEWS) and the San Francisco syncope score (SFSS) were determined in every patient. Primary endpoint was mortality after 180 days.Eight patients (2%) died after 180 days. DC was significantly lower in the group of nonsurvivors as compared with survivors (3.1 ±â€Š2.5 ms vs 6.7 ±â€Š2.4 ms; P < .001), whereas the MEWS was comparable in both was comparable in both groups. (2.1 ±â€Š0.8 vs 2.1 ±â€Š1.0; P = .84). The SFSS failed at identifying 4 of 8 nonsurvivors (50%) as high risk patients. No patient with a favorable DC (≥7 ms) died (0.0% vs 3.7%; P = .01, OR 0.55 (95% CI 0.40-0.76), P < .001). In the receiver operating characteristic (ROC) analysis DC yielded an area under the curve of 0.85 (95% CI 0.71-0.98).Our study demonstrates that DC is a predictor of 180-days-mortality in patients admitted to the ED due to syncope. Syncope patients at low risk can be identified by DC and may be discharged safely.

[Mechanic valve prosthesis and pregnancy: Is Phenprocoumon replaceable?]. / Kunstklappe in der Schwangerschaft: Ist Phenprocoumon ersetzbar? - Fall 2 / 2016.

HISTORY AND ADMISSION FINDINGS: We report the case of a 30-year-old pregnant patient with mechanical valve replacement in mitral and aortic position. She had discontinued Phenprocoumon-treatment in the 5+4 week of pregnancy by herself. Because of rheumatic fever she had undergone a mechanical aortic and mitral valve replacement 12 years ago. Due to a thrombosis of the mitral valve, an acute reoperation had to be done 5 years later. 2 years ago, a partially re-thrombosis of the mechanical mitral valve was treated by intravenous thrombolysis. These complications had been probably due to incomplicance. The patient had experienced 3 abortions before. INVESTIGATIONS: The vaginal sonography determined an intact gestation. The laboratory test revealed an INR of 1.2. The transesophageal echocardiography showed a partially thrombosed mechanical mitral valve. The abdominal ultrasonography detected an embolic splenic infarction. DIAGNOSIS, TREATMENT AND CLINICAL COURSE: These findings were consistent with partially thrombosed mechanical mitral valve with thromboembolic splenic infarction among incompetent oral anticoagulation. After initial heparinization with under twice daily control of the partial thromboplastin time the joint decision was made to restart Phenprocoumon (target INR 2.5 to 3.5, and additional ASS 100 mg /day). 9 days later the patient had a missed abortion. An uncomplicated curettage was performed under therapeutic i.v. heparinization. CONCLUSIONS: The use of coumarins in pregnancy carries a fetal risk. But it is the most secure anticoagulation after a mechanical valve replacement, especially in high-risk patients. Alternatives are heparins. They don't cross the placenta but are associated with a slightly elevated risk of thromboembolism.

Cargo-selective apical exocytosis in epithelial cells is conducted by Myo5B, Slp4a, Vamp7, and Syntaxin 3.

Mutations in the motor protein Myosin Vb (Myo5B) or the soluble NSF attachment protein receptor Syntaxin 3 (Stx3) disturb epithelial polarity and cause microvillus inclusion disease (MVID), a lethal hereditary enteropathy affecting neonates. To understand the molecular mechanism of Myo5B and Stx3 interplay, we used genome editing to introduce a defined Myo5B patient mutation in a human epithelial cell line. Our results demonstrate a selective role of Myo5B and Stx3 for apical cargo exocytosis in polarized epithelial cells. Apical exocytosis of NHE3, CFTR (cystic fibrosis transmembrane conductance regulator), and GLUT5 required an interaction cascade of Rab11, Myo5B, Slp4a, Munc18-2, and Vamp7 with Stx3, which cooperate in the final steps of this selective apical traffic pathway. The brush border enzymes DPPIV and sucrase-isomaltase still correctly localize at the apical plasma membrane independent of this pathway. Hence, our work demonstrates how Myo5B, Stx3, Slp4a, Vamp7, Munc18-2, and Rab8/11 cooperate during selective apical cargo trafficking and exocytosis in epithelial cells and thereby provides further insight into MVID pathophysiology.