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Signaling by the Ral small GTPase is poorly understood in vivo . Caenorhabditis elegans animals with constitutively activated RAL-1 or deficient for the inhibitory RalGAP, HGAP-1 /2, display pale intestines. Staining with Oil Red O detected decreased intestinal lipids in the hgap-1 deletion mutant relative to the wild type. Constitutively activated RAL-1 decreased lipid detected by stimulated Raman scattering (SRS) microscopy, a label-free method of detecting lipid by laser excitation and detection. A signaling-deficient missense mutant for RAL-1 also displayed reduced lipid staining via SRS. We conclude that RAL-1 signaling regulates lipid homeostasis, biosynthesis or storage in live animals.

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Background: Multiple endocrine neoplasia type 2B (MEN 2B) is a rare hereditary syndrome caused mainly by Met918Thr germline RET mutation and characterized by medullary thyroid carcinoma (MTC), pheochromocytoma (PHEO), and typical phenotypic features. MEN 2B cases previously reported in the literature have variable clinical course. Objectives: We aimed to discuss the characteristics of four MEN 2B cases with unusual presentations,clinical course and review the recent clinical data on MEN2B. Results: All patients had de novo M918T mutation and no family history. The mean age of patients was 38.2 years (27-56). Two patients had typical phenotypic features of MEN 2B; the other two patients had no striking phenotypic features. First detected MEN 2B component was MTC in two, intestinal ganglioneuromatosis in one, and PHEO in one of the cases. Bilateral PHEO was detected in all four cases. Conclusions: MEN 2B is a complex syndrome characterized by wide phenotypic variability and different clinical outcomes. To diagnose sporadic MEN 2B cases, genetic testing should be performed in all cases with suspicious clinical features. Although early diagnosis is the main factor that increases life expectancy, some MEN 2B patients with late diagnosis may exhibit a mild clinical course and better prognosis than expected, with effective treatment.

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Esophageal squamous cell carcinoma (ESCC) poses a major challenge for clinicians as the prognosis is poor and treatment options are limited. However, recent advances in immunotherapy have significantly changed the treatment algorithm of ESCC. Patients with early ESCC should undergo an endoscopic resection. If histological margins are infiltrated with tumor cells or other risk factors for lymph node metastasis are present, further resective surgery should be offered. In a locally advanced setting, radiochemotherapy with or without resection remains the standard of care. In the absence of pathological complete response after neoadjuvant radiochemotherapy and R0 resection, adjuvant immunotherapy for 1 year should be administered to improve disease-free survival. In metastatic first-line setting, combination of platin/fluoropyrimidine-based systemic chemotherapy with checkpoint inhibitors is the novel standard of care for all-comers in the United States and for patients with programmed death-ligand 1 positivity in Europe. Immunotherapy has also been approved in a second-line setting. However, the benefit from immunotherapy reinduction is still unknown and, therefore, standard second-line chemotherapy with taxanes or irinotecan is still the treatment of choice after progression on immunochemotherapy. It is of highest importance that treatment decisions are based on informed patient wishes and are discussed in an interdisciplinary tumor board. This review summarizes how to manage, in our opinion, patients with ESCC and gives a practical overview of the treatment strategies in Europe.

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Obtaining efficient autotrophic ammonia removal (aka partial nitritation-anammox, or PNA) requires a balanced microbiome with abundant aerobic and anaerobic ammonia oxidizing bacteria and scarce nitrite oxidizing bacteria. Here, we analyzed the microbiome of an efficient PNA process that was obtained by sequential feeding and periodic aeration. The genomes of the dominant community members were inferred from metagenomes obtained over a 6 month period. Three Brocadia spp. genomes and three Nitrosomonas spp. genomes dominated the autotrophic community; no NOB genomes were retrieved. Two of the Brocadia spp. genomes lacked the genomic potential for nitrite reduction. A diverse set of heterotrophic genomes was retrieved, each with genomic potential for only a fraction of the denitrification pathway. A mutual dependency in amino acid and vitamin synthesis was noted between autotrophic and heterotrophic community members. Our analysis suggests a highly-reticulated nitrogen cycle in the examined PNA microbiome with nitric oxide exchange between the heterotrophs and the anammox guild.

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BACKGROUND: Gastroesophageal adenocarcinoma is a major contributor to global disease burden with poor prognosis even in resectable, regionally limited stages. Feasible prognostic tools are crucial to improve patient management, yet scarce. PATIENTS AND METHODS: Disease-related symptoms, patient, tumour, treatment as well as laboratory parameters at initial diagnosis and overall survival (OS) of patients with stage II and III gastroesophageal adenocarcinoma, who were treated between 1990 and 2020 at the Medical University of Vienna, were evaluated in a cross-validation model to develop a feasible risk prediction score. RESULTS: In total, 628 patients were included in this single-centre analysis. The final score ranked from 0 to 10 and included the factors sex (female +1), age, years (30-59 +1, >60 +2), underweight classified by body mass index (+2), location of the tumour (stomach +1), stage (III +2), stenosis in endoscopy (+1) and weight loss (+1). The score was grouped into low- (0-3), medium- (4-6) and high-risk (7+) subgroups. The median OS were 70.3 [95% confidence interval (CI) 51.2-111.8], 23.4 (95% CI 21.2-26.7) and 12.6 (7.0-16.1) months, respectively. The 1-year survival probabilities were 0.88 (95% CI 0.83-0.93), 0.75 (95% CI 0.70-0.79) and 0.54 (95% CI 0.39-0.74), whereas the 5-year survival probabilities were 0.57 (95% CI 0.49-0.66), 0.24 (95% CI 0.20-0.28) and 0.09 (95% CI 0.03-0.28), respectively. CONCLUSIONS: The VIennese risk prediction score for Oesophagogastric Localized Adenocarcinoma (VIOLA) risk prediction score poses a feasible tool for the estimation of OS in patients with regionally limited gastroesophageal adenocarcinoma and, thus, may improve patient management in clinical routine. Prospective analyses should be carried out to confirm our findings.

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BACKGROUND: The aim of this study was to assess the in vitro fertilization (IVF) treatments outcomes of women with recurrent implantation failure (RIF) after hysteroscopic metroplasty for T-shaped uterus. METHODS: This retrospective, observational study included 85 patients with RIF and T-shaped uterus who presented to In Vitro Fertilization Unit of Hüma Obstetrics and Gynecology Hospital between January, 2018 and August, 2021. The hysteroscopic metroplasty was performed before IVF procedure in 43 of patients included. Remaining 42 patents underwent IVF without hysteroscopic metroplasty were included as controls. The fertility outcome were compared between groups. RESULTS: After 3 months, spontaneous pregnancy was achieved in 10.4% (5/48) of patients with T-shaped uterus underwent hysteroscopic metroplasty. In the hysteroscopy group, implantation rate, clinical pregnancy rate, biochemical pregnancy rate and the miscarriage rate were 55.8% (24/43), 41.9% (18/43), 14% (6/43) and 11.6% (5/43, respectively. The live birth rate after in vitro fertilization was 30.2% (13/43) in the hysteroscopy group and 9.5% (4/42) in the control group (p<0.05). There were no significant differences in the IVF outcomes such as the biochemical pregnancy rate and miscarriage rate between the hysteroscopy and control groups. CONCLUSION: This study demonstrates that hysteroscopic metroplasty improves pregnancy and live birth rates for women with a history of RIF and dysmorphic uterus. Correction of T-shaped uterus with hysteroscopic metroplasty ensured favorable reproductive outcomes in RIF cases. However, conclusions must be taken carefully as this is an observational study. A prospective, randomized and controlled study is necessary to support these results.

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OBJECTIVE@#To assess shear bond strength and failure mode (Adhesive Remnant Index, ARI) of orthodontic brackets bonded to polymethylmethacrylate (PMMA) blocks for computer-aided design/manufacture (CAD/CAM) fabrication of temporary restorations, following substrate chemical or mechanical treatment.@*METHODS@#Two types of PMMA blocks were tested: CAD-Temp® (VITA) and Telio® CAD (Ivoclar-Vivadent). The substrate was roughened with 320-grit sandpaper, simulating a fine-grit diamond bur. Two universal adhesives, Scotchbond Universal Adhesive (SU) and Assure Plus (AP), and a conventional adhesive, Transbond XT Primer (XTP; control), were used in combination with Transbond XT Paste to bond the brackets. Six experimental groups were formed: (1) CADTemp®/SU; (2) CAD-Temp®/AP; (3) CAD-Temp®/XTP; (4) Telio® CAD/SU; (5) Telio® CAD/AP; (6) Telio® CAD/XTP. Shear bond strength and ARI were assessed. On 1 extra block for each PMMA-based material surfaces were roughened with 180-grit sandpaper, simulating a normal/medium-grit (100 mm) diamond bur, and brackets were bonded. Shear bond strengths and ARI scores were compared with those of groups 3, 6.@*RESULTS@#On CAD-Temp® significantly higher bracket bond strengths than on Telio® CAD were recorded. With XTP significantly lower levels of adhesion were reached than using SU or AP. Roughening with a coarser bur resulted in a significant increase in adhesion.@*CONCLUSIONS@#Bracket bonding to CAD/CAM PMMA can be promoted by grinding the substrate with a normal/medium-grit bur or by coating the intact surface with universal adhesives. With appropriate pretreatments, bracket adhesion to CAD/CAM PMMA temporary restorations can be enhanced to clinically satisfactory levels.

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Attempts for identifying targeted therapy strategies in metastatic gastric and gastroesopheal junction cancer (upper-GI) revealed that the inhibition of human epidermal growth factor receptor-2 (HER2) by monoclonal antibody trastuzumab improves survival of these patients. Hence, adding trastuzumab to doublet chemotherapy has become the standard treatment in this setting. Although the patient survival is extended among clinical trials, the knowledge on the real-time setting is limited. With this retrospective, single center analysis of the patient data of the Medical University of Vienna, we sought to investigate the clinical characteristics and outcome of patients, who received trastuzumab-based chemotherapy for metastatic upper-GI tumor. All patients, who received trastzumab at least once were included to the analysis. Clinical and pathological data were recorded. This search revealed 33 patients. The demographic data was comparable with that of the previous clinical trials. Progression free survival (PFS) was 11 months, whereas overall survival (OS) was 21 months. OS was significantly associated with initially favorable response to treatment. Thirteen patients (39%) received trastuzumab as maintenance treatment with a median cycle number of 6. Toxicity profile was acceptable with only one patient detected to have cardiotoxicity. Taken together, trastuzumab based treatment induced a considerable PFS and OS in metastatic or advanced upper-GI tumors with acceptable toxicity profile. The maintenance therapy with trastuzumab was safe and effective in patients who had initially a favorable response to chemotherapy. The optimal duration of the maintenance therapy should be tested in future clinical trials.

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Several neurodevelopmental diseases are characterized by impairments in cortical morphology along with altered white matter connectivity. However, the relationship between these two measures is not yet clear. In this study, we propose a novel methodology to compute and display metrics of white matter connectivity at each cortical point. After co-registering the extremities of the tractography streamlines with the cortical surface, we computed two measures of connectivity at each cortical vertex: the mean tracts' length, and the proportion of short- and long-range connections. The proposed measures were tested in a clinical sample of 62 patients with 22q11.2 deletion syndrome (22q11DS) and 57 typically developing individuals. Using these novel measures, we achieved a fine-grained visualization of the white matter connectivity patterns at each vertex of the cortical surface. We observed an intriguing pattern of both increased and decreased short- and long-range connectivity in 22q11DS, that provides novel information about the nature and topology of white matter alterations in the syndrome. We argue that the method presented in this study opens avenues for additional analyses of the relationship between cortical properties and patterns of underlying structural connectivity, which will help clarifying the intrinsic mechanisms that lead to altered brain structure in neurodevelopmental disorders.

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Emericella nidulans is a species that has only rarely been implicated in human disease after cataract surgery. Here, we report the first postoperative case in the literature, as far as we know. The patient was a 50-year-old patient presented with mild anterior uveitis one week after cataract surgery, and hypopion developed over the next two days. First microbiological evaluation and the results of direct microscopy and cultures of the anterior chamber and vitreous samples were found to be negative. Despite vigorous topical and intravitreal (vancomycin and amikacin) therapy, the endophthalmitis did not improve. Anterior chamber paracentesis, vitreous tap and finally complete vitrectomy with removal of the capsular bag including the intraocular lens (IOL) were performed. The anterior chamber, vitreous fluid samples and IOL were submitted to the microbiology laboratory: the culture yielded E. nidulans growth. Ocular inflammation resolved and vision improved on intravenous, subconjunctival and long-term oral voriconazole treatment. E. nidulans can be an important cause of ocular fungal infections including endophthalmitis, and voriconazole seems to be effective for the treatment of E. nidulans endophthalmitis.

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Autotrophic nitrogen removal is regarded as a resource efficient process to manage nitrogen-rich residual streams. However, nitrous oxide emissions of these processes are poorly documented and strategies to mitigate emissions unknown. In this study, two sequencing batch reactors performing single-stage nitritation/anammox were operated under different aeration strategies, gradually adjusted over six months. At constant but limiting oxygen loading, synthetic reject water was fed (0.75 g-N/L · d) and high nitrogen removal efficiencies (83 ± 5 and 88 ± 2%) obtained. Dynamics of liquid phase nitrous (N2O) and nitric oxide (NO) concentrations were monitored and N2O emissions calculated. Significant decreases in N2O emissions were obtained when the frequency of aeration was increased while maintaining a constant air flow rate (from >6 to 1.7% ΔN2O/ΔTN). However, no significant effect on the emissions was noted when the duration of aeration was increased while decreasing air flow rate (10.9 ± 3.2% ΔN2O/ΔTN). The extant ammonium oxidation activity (mgNH4(+)-N/gVSS · min) positively correlated with the specific N2O production rate (mgN2O-N/gVSS · min) of the systems. Operating under conditions where anaerobic exceeds aerobic ammonium oxidation activity is proposed to minimize N2O emissions from single-stage nitritation/anammox reactors; increasing the frequency of aeration cycling is an efficient way of obtaining those conditions.

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OBJECTIVE: There is increasing evidence that congenital heart disease (CHD) affects brain structure, but little is known about the long-term trajectory of brain maturation and its impact on the cognitive development of patients with CHD. We proposed to address this question in a longitudinally-followed cohort of individuals with 22q11.2 deletion syndrome (22q11DS), the most common microdeletion syndrome in humans. METHODS: A total of 80 participants were included in this longitudinal analysis. The volumes of thirty-four cortical regions and eight hippocampal regions were measured in each hemisphere with FreeSurfer software. This paper utilized linear mixed modelling to investigate cerebral morphometry and age-related maturational changes of all regions. The effect of CHD was assessed for intercept and slope significance. RESULTS: We observed significant (p < .05/34) volumetric reductions in patients with CHD compared to patients without in fifteen out of the sixty-eight cortical sub-regions. Similarly, global hippocampal volumes and twelve of the hippocampal sub-regions were significantly smaller (p < .05/8). The results demonstrate significant absolute volumetric differences, but did not show any significant differences in the way the cortical or hippocampal regions developed over time. There was limited evidence of any effect of the presence of CHD on key cognitive measures. CONCLUSIONS: We propose that cerebral hypoperfusion, due to the presence of CHD or its surgery, impairs early cortical and particularly hippocampal growth, potentially due to the damaging effects of stress, but not subsequent maturational processes in children and adolescents.

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BACKGROUND: Survival upon diagnosis of brain metastases (BM) in patients with non-small cell lung cancer (NSCLC) is highly variable and established prognostic scores do not include tissue-based parameters. METHODS: Patients who underwent neurosurgical resection as first-line therapy for newly diagnosed NSCLC BM were included. Microvascular density (MVD), Ki67 tumor cell proliferation index and hypoxia-inducible factor 1 alpha (HIF-1 alpha) index were determined by immunohistochemistry. RESULTS: NSCLC BM specimens from 230 patients (151 male, 79 female; median age 56 years; 199 nonsquamous histology) and 53/230 (23.0%) matched primary tumor samples were available. Adjuvant whole-brain radiation therapy (WBRT) was given to 153/230 (66.5%) patients after neurosurgical resection. MVD and HIF-1 alpha indices were significantly higher in BM than in matched primary tumors. In patients treated with adjuvant WBRT, low BM HIF-1 alpha expression was associated with favorable overall survival (OS), while among patients not treated with adjuvant WBRT, BM HIF-1 alpha expression did not correlate with OS. Low diagnosis-specific graded prognostic assessment score (DS-GPA), low Ki67 index, high MVD, low HIF-1 alpha index and administration of adjuvant WBRT were independently associated with favorable OS. Incorporation of tissue-based parameters into the commonly used DS-GPA allowed refined discrimination of prognostic subgroups. CONCLUSION: Ki67 index, MVD and HIF-1 alpha index have promising prognostic value in BM and should be validated in further studies.

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22q11.2 deletion syndrome (22q11DS) is associated with increased risk for schizophrenia. Better identifying risk factors for the emergence of psychotic symptoms in this population is needed to improve clinical assessment and early interventions. Schizophrenia spectrum disorders, hallucinations and delusions were characterized in an original sample of 104 individuals with 22q11DS. Further analysis of positive and negative symptoms was performed in a subsample of 59 individuals. Finally, longitudinal data available in 56 patients were used to explore the developmental trajectories of psychotic symptoms as well as the associations between psychotic symptoms and cognitive functioning. Schizophrenia spectrum disorders and psychotic symptoms were frequent in adolescent and adults with 22q11DS. The severity of hallucinations and non-persecutory delusional ideas discriminated patients at ultra-high risk for conversion to psychosis. Whereas approximately one-third of patients experienced an emergence of psychotic symptoms during a 4-year interval, 20 % displayed transient symptoms. Individuals with psychotic symptoms were characterized by a lower cognitive functioning in the context of the 22q11DS. The present study adds important data on the characteristics and developmental trajectory of psychotic symptoms in this population. This information may ultimately help clinicians dealing with these patients to reduce the duration of untreated psychosis and improve outcome.

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Start-up and operation of single-stage nitritation-anammox sequencing batch reactors (SBRs) for completely autotrophic nitrogen removal can be challenging and far from trivial. In this study, a step-wise procedure is developed based on stoichiometric analysis of the process performance from nitrogen species measurements to systematically guide start-up and normal operation efforts (instead of trial and error). The procedure is successfully applied to laboratory-scale SBRs for start-up and maintained operation over an 8-month period. This analysis can serve as a strong decision-making tool to take appropriate actions with respect to reactor operation to accelerate start-up or ensure high-rate N removal via the nitritation-anammox pathway.

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Extracellular polymeric substances (EPS) have a presumed determinant role in the structure, architecture, strength, filterability, and settling behaviour of microbial solids in biological wastewater treatment processes. Consequently, numerous EPS extraction protocols have recently been published that aim to optimize the trade off between high EPS recovery and low cell lysis. Despite extensive efforts, the obtained results are often contradictory, even when analysing similar biomass samples and using similar experimental conditions, which greatly complicates the selection of an extraction protocol. This study presents a rigorous and critical assessment of existing physical and chemical EPS extraction methods applied to mixed-culture biomass samples (nitrifying, nitritation-anammox, and activated sludge biomass). A novel fluorescence-based method was developed and calibrated to quantify the lysis potential of different EPS extraction protocols. We concluded that commonly used methods to assess cell lysis (DNA concentrations or G6PDH activities in EPS extracts) do not correlate with cell viability. Furthermore, we discovered that the presence of certain chemicals in EPS extracts results in severe underestimation of protein and carbohydrate concentrations by using standard analytical methods. Keeping both maximum EPS extraction yields and minimal biomass lysis as criteria, it was identified a sonication-based extraction method as the best to determine and compare tightly-bound EPS fractions in different biomass samples. Protein was consistently the main EPS component in all analysed samples. However, EPS from nitrifying enrichments was richer in DNA, the activated sludge EPS had a higher content in humic acids and carbohydrates, and the nitritation-anammox EPS, while similar in composition to the nitrifier EPS, had a lower fraction of hydrophobic biopolymers. In general, the easily-extractable EPS fraction was more abundant in carbohydrates and humic substances, while DNA could only be found in tightly bound EPS fractions. In conclusion, the methodology presented herein supports the rational selection of analytical tools and EPS extraction protocols in further EPS characterization studies.

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While significant differences in male and female brain structures have commonly been reported, only a few studies have focused on the sex differences in the way the cortex matures over time. Here, we investigated cortical thickness maturation between the age of 6 to 30 years, using 209 longitudinally-acquired brain MRI scans. Significant sex differences in the trajectories of cortical thickness change with age were evidenced using non-linear mixed effects models. Similar statistical analyses were computed to quantify the differences between cortical gyrification changes with age in males and females. During adolescence, we observed a statistically significant higher rate of cortical thinning in females compared to males in the right temporal regions, the left temporoparietal junction and the left orbitofrontal cortex. This finding is interpreted as a faster maturation of the social brain areas in females. Concomitantly, statistically significant sex differences in cortical folding changes with age were observed only in one cluster of the right prefrontal regions, suggesting that the mechanisms underlying cortical thickness and gyrification changes with age are quite distinct. Sexual dimorphism in the developmental course of the cortical maturation may be associated with the different age of onset and clinical presentation of many psychiatric disorders between males and females.

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UNLABELLED: We aimed to investigate the effect of allograft amniotic membrane utilization to prevent the post-operative adhesion formation. BACKGROUND: In 24 pregnant inbreed Wistar-Albino rats, pregnancy was terminated by forming bilateral uterine horn defect via cesarean section at 20th gestational day. Rats were assigned in three groups randomly. METHODS: In the first group, abdomen closure was achieved without administration of any intra-peritoneal material following standard surgical intervention. In the second group Seprafilm was used to cover the defect at anterior horn of uterus; whereas amniotic membrane of the rat itself was used in the third group. After 3 weeks, all rats were sacrificed and re-laparotomy was performed to determine adhesions scores. RESULTS: No significant difference was found in adhesion scores between the group 1 and group 3 and also between group 2 and group 3, supporting the previous findings in the literature. CONCLUSION: We observed that direct application of allograft amniotic membrane, which is an adhesion barrier used after cesarean section, to injured surface had no effect in the prevention of adhesions (Tab. 3, Fig. 5, Ref. 28).

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In many animals, rhythmic motor activity is governed by neural limit cycle oscillations under the control of sensory feedback. In the fruit fly Drosophila melanogaster, the wingbeat rhythm is generated myogenically by stretch-activated muscles and hence independently from direct neural input. In this study, we explored if generation and cycle-by-cycle control of Drosophila's wingbeat are functionally separated, or if the steering muscles instead couple into the myogenic rhythm as a weak forcing of a limit cycle oscillator. We behaviourally tested tethered flying flies for characteristic properties of limit cycle oscillators. To this end, we mechanically stimulated the fly's 'gyroscopic' organs, the halteres, and determined the phase relationship between the wing motion and stimulus. The flies synchronized with the stimulus for specific ranges of stimulus amplitude and frequency, revealing the characteristic Arnol'd tongues of a forced limit cycle oscillator. Rapid periodic modulation of the wingbeat frequency prior to locking demonstrates the involvement of the fast steering muscles in the observed control of the wingbeat frequency. We propose that the mechanical forcing of a myogenic limit cycle oscillator permits flies to avoid the comparatively slow control based on a neural central pattern generator.

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PURPOSE: Previous functional neuroimaging studies showed that the brainstem may have an important role in migraine and recently, DTI studies demonstrated that structural alterations in migraineurs may extend beyond the normal appearing brain tissue. The aim of our study was to find out if DTI may detect any abnormality during the spontaneous migraine attacks. METHODS: The DTI images obtained in a 3T system during spontaneous migraine episodes. Patients with any systemic or metabolic disorder and abnormal signal intensity in conventional sequences were excluded. We measured the FA and ADC values of red nuclei, periaquaductal gray matter, thalami, posterior limbs of internal capsules and subcortical white matter. Fifteen healthy volunteers served as control group. RESULTS: Fourteen patients enrolled in the study. The only site where we found an abnormality was the red nuclei, where the ADC values in migraineurs were statistically higher than in healthy volunteers. There was no statistical correlation between the DTI measurements and patients' ages, duration of disease, frequency of attacks and localization of pain. CONCLUSION: Our findings supported the findings of previous functional neuroimaging studies, which concluded that the brainstem might have a role in the pathogenesis of a migraine episode. We think that the increase of ADC values in red nuclei may reflect vasogenic edema, which cannot be detected in conventional sequences. However, the exact underlying mechanism for this observation is unclear and we do not know whether these changes are responsible for triggering an attack or if they are the consequents of the attack itself.

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