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A method is presented to generate quantitative precipitation estimates over Alaska using kriging to merge sparse, unevenly distributed rain gauge observations with quantitative precipitation forecasts from a three-member ensemble of high-resolution numerical weather prediction models. The estimated error variance of the analysis is computed by starting with the estimated error variance from kriging and then refining the variance in k-fold cross validation by an empirically derived inflation factor. The method combines dynamical model forecast information with observational data to deliver a best linear unbiased estimate of precipitation, along with an analysis uncertainty estimate, that provides a much-needed precipitation analysis in a region where sparse in situ observations, poor coverage by remote sensing platforms, and complex terrain introduce large uncertainties that need to be quantified. For 6-hourly accumulation estimates produced four times daily from 1 August 2019 to 31 July 2020, three analysis configurations are tested to measure the value added by including model forecast data and how those data are best utilized in the analysis. Several directions for further improvement and validation of the analysis product are provided.

Purpose Autosomal dominant polycystic kidney disease (ADPKD) is a common hereditary disorder and accounts for 5–10% of all cases of kidney failure. 50% of ADPKD patients reach kidney failure by the age of 58 years requiring dialysis or transplantation. Nephrectomy is performed in up to 20% of patients due to compressive symptoms, renal-related complications or in preparation for kidney transplantation. However, due to the large kidney size in ADPKD, nephrectomy can come with a considerable burden. Here we evaluate our institution’s experience of laparoscopic nephrectomy (LN) as an alternative to open nephrectomy (ON) for ADPKD patients. Materials and methods We report the results of the first 12 consecutive LN for ADPKD from August 2020 to August 2021 in our institution. These results were compared with the 12 most recent performed ON for ADPKD at the same institution (09/2017 to 07/2020). Intra- and postoperative parameters were collected and analyzed. Health related quality of life (HRQoL) was assessed using the SF36 questionnaire. Results Age, sex, and median preoperative kidney volumes were not significantly different between the two analyzed groups. Intraoperative estimated blood loss was significantly less in the laparoscopic group (33 ml (0–200 ml)) in comparison to the open group (186 ml (0–800 ml)) and postoperative need for blood transfusion was significantly reduced in the laparoscopic group (p = 0.0462). Operative time was significantly longer if LN was performed (158 min (85–227 min)) compared to the open procedure (107 min (56–174 min)) (p = 0.0079). In both groups one postoperative complication Clavien Dindo ≥ 3 occurred with the need of revision surgery. SF36 HRQol questionnaire revealed excellent postoperative quality of life after LN. Conclusion LN in ADPKD patients is a safe and effective operative procedure independent of kidney size with excellent postoperative outcomes and benefits of minimally invasive surgery. Compared with the open procedure patients profit from significantly less need for transfusion with comparable postoperative complication rates. However significant longer operation times need to be taken in account.

Objective: Our objective was to evaluate the accuracy of the prenatal diagnosis and the relation between the type of right aortic arch (RAA) with other intra- or extracardiac (EC) and chromosomal anomalies. Methods: A retrospective, observational study was conducted between 2011–2020 in a Romanian tertiary center. All RAA cases, including double aortic arch (DAA), were extracted from the databases and studied thoroughly. Results: We detected 18 RAA cases: five (27.78%) type I (mirror image, “V” type), 11 (61.12%) type II (“U” type), and two (11.10%) DAA cases. Heart anomalies were associated in 38.89% (overall), 60% (type I), 36.37% (type II), and 0% (DAA) cases. Tetralogy of Fallot represented the most prevalent cardiac malformation (in 22.23% of cases). EC anomalies were present in 44.44% of fetuses (20% of type I, 54.55% of type II, and 50% of DAA cases). Genetic abnormalities were found in 41.17% of pregnancies, with 22q11.2 deletion in 23.53%. 55.55% of the cases had a good neonatal evolution and 44.45% of the pregnancies were terminated. An overall good outcome of pregnancy was noted in 40% of type I RAA, 63.64% of type II RAA, and 50% of DAA cases. All RAA cases examined in the first trimester were correctly diagnosed. Conclusions: RAA can be accurately diagnosed and classified by means of prenatal ultrasound since early pregnancy. A detailed anatomy scan and genetic testing, including 22q11 deletion, should be offered to all pregnancies when RAA is discovered. When isolated, RAA associates a good outcome, indifferently the anatomical type.

Autosomal dominant polycystic kidney disease (ADPKD) is a common hereditary disorder and accounts for 5-10% of all cases of kidney failure. 50% of ADPKD patients reach kidney failure by the age of 58 years requiring dialysis or transplantation. Nephrectomy is performed in up to 20% of patients due to compressive symptoms, renal-related complications or in preparation for kidney transplantation. However, due to the large kidney size in ADPKD, nephrectomy can come with a considerable burden. Here we evaluate our institution's experience of laparoscopic nephrectomy (LN) as an alternative to open nephrectomy (ON) for ADPKD patients.We report the results of the first 12 consecutive LN for ADPKD from August 2020 to August 2021 in our institution. These results were compared with the 12 most recent performed ON for ADPKD at the same institution (09/2017 to 07/2020). Intra- and postoperative parameters were collected and analyzed. Health related quality of life (HRQoL) was assessed using the SF36 questionnaire.Age, sex, and median preoperative kidney volumes were not significantly different between the two analyzed groups. Intraoperative estimated blood loss was significantly less in the laparoscopic group (33 ml (0-200 ml)) in comparison to the open group (186 ml (0-800 ml)) and postoperative need for blood transfusion was significantly reduced in the laparoscopic group (p = 0.0462). Operative time was significantly longer if LN was performed (158 min (85-227 min)) compared to the open procedure (107 min (56-174 min)) (p = 0.0079). In both groups one postoperative complication Clavien Dindo ≥ 3 occurred with the need of revision surgery. SF36 HRQol questionnaire revealed excellent postoperative quality of life after LN.LN in ADPKD patients is a safe and effective operative procedure independent of kidney size with excellent postoperative outcomes and benefits of minimally invasive surgery. Compared with the open procedure patients profit from significantly less need for transfusion with comparable postoperative complication rates. However significant longer operation times need to be taken in account.

Ovarian ectopic pregnancy (OEP) represents the rarest type of ectopic pregnancy, accounting for 1-3% of this pathology. The diagnosis of this pathology is challenging due to the non-specific clinical aspects and the ultrasound examination hampered by the lack of visible gestational sac in the presence of hematocele and hemoperitoneum. The purpose of the extended histopathological (HP) examination was to identify particular aspects of the OEP trophoblast and to highlight potential local ovarian modifications which can determine pregnancy fixation at this level. The patient presented local favorable conditions for intraovarian nidation, conditions confirmed by the HP classical examination and by the immunohistochemical evaluation. We identified, using classical Hematoxylin-Eosin, Masson's trichrome and Periodic Acid-Schiff (PAS)-Hematoxylin, necrotic hemorrhage, accentuated vascular thrombosis and high density lymphoplasmocytary infiltrate. These modifications increased local adhesivity and cell destruction through hypoperfusion. Anti-cluster of differentiation antibodies (CD34, CD38, tryptase) revealed the low number of intravillous vessels and the high number of macrophages and mastocytes involved in the local inflammatory process heighten. We identified the presence of trophoblast tissue in the ovarian structure using anti-cytokeratin AE1∕AE3 (CK AE1∕AE3)/anti-cytokeratin 7 (CK7) antibodies. The anti-alpha-smooth muscle actin (α-SMA) and anti-vimentin (VIM) antibodies displayed the density of myofibroblasts and intravillous stromal cells and with the aid of anti-progesterone receptor (PR) antibody, we identified the corpus luteum hormonal response in the OEP. The placental villosities present a blocked multiplication process at the anti-apoptotic B-cell lymphoma 2 (BCL2) protein, confirmed by the Ki67 cell proliferation and tumor protein 63 (p63) immunomarkers. Anti-neuron specific enolase (NSE), anti-calretinin and anti-inhibin A antibodies showed the particular aspects of the granulosa and internal theca cells, which may be involved in oocyte release blockage, intraluteal and extraluteal fecundation of the OEP.

(1) Background: Neurological Soft Signs (NSS) are subtle neurological abnormalities that are more common in schizophrenia patients than in healthy individuals and have been regularly observed in neuroleptic-naive first-episode patients, supporting the hypothesis that they are an intrinsic component of schizophrenia. (2) Methods: a review of articles published in the last ten years (from January 2013 to January 2023) was carried out on articles published in ScienceDirect and PubMed, by following the PRISMA Statement extension for scoping reviews (PRISMA-ScR), which evaluated the impact of NSS in correlation with the symptomatology, neuroleptic treatment, and the cerebral structural changes of patients with schizophrenia. (3) Results: thirty articles were included, among them twelve included MRI structural evaluation and four studies with a longitudinal design. (4) Conclusions: interest in researching NSS has increased in recent years, but questions remain about their origin and relationship to schizophrenia symptoms, thus this study aims to fill in information gaps in the hope that future research will help provide individualized treatment. It is suggested that NSS in schizophrenia might have an inherited genetic relationship pattern, thus being in line with a trait viewpoint. Most of the research revealed that schizophrenia patients had higher NSS scores than healthy controls, however, they were rather similar to their first-degree relatives, thus, also arguing in favor of a trait perspective. The greatest improvement in scores is seen in those with a remitting course, as shown by declining NSS ratings concurrent with symptomatology.

OBJECTIVE In this pilot study, we tested the feasibility of cardiac structures reconstruction from histological sections in 12-13 weeks normal fetuses. Conventional autopsy is hampered at this gestational age because of the small size of the heart anatomical structures, while alternative non-invasive methods for pathology examination of the fetus are expensive, rarely available and lack accuracy data regarding the confirmation of first trimester heart defects suspected by early prenatal ultrasound (US) scans. MATERIALS AND METHODS Normal hearts from fetuses aged 12-13 gestational weeks (GW) were harvested for histological preparation, virtual reconstruction, and cardiac structures analysis. The normalcy of heart structures was confirmed before pregnancy termination, using a detailed US scan protocol. The fetal heart was routinely processed for formalin fixation and paraffin embedding (FFPE) and 10 μm seriate sections have been cut until finishing the specimen. All sections have been scanned and a three-dimensional (3D) reconstruction of the whole organ has been rendered, based on computer-aided manual tracing. Using the 3D navigation software, the main cardiac structures were searched for a proper and confident visualization. RESULTS Five cases were investigated. Visualization of the normal heart cavities, including atrioventricular septum was very good in all fetuses. The entire course of right and left ventricle outflow tracts was confidently confirmed, along the branching pattern of aorta and pulmonary artery trunk. Regarding the veno-atrial connections, it was easy to identify the entrance of the inferior and superior caval veins into the right atrium, but a detailed review of the histological sections was necessary for the visualization of the left atrium venous openings. The inherent morphological deformation following heart block sectioning resulted in a lower resolution or quality of the "reconstructed" planes, but these distortions did not represent a significant impediment in any of the cases. The resources involved ordinary histology and information technology (IT) equipment. To further decrease the time involved by the protocol, many steps may be automated: cutting, coloring, and scanning. CONCLUSIONS The results indicate that this method can be implemented to routine clinical practice. The use of 3D reconstruction of fetal heart histological sections in first trimester may serve as an important audit to confirm the normalcy of heart structures. Also, the histological and postprocessed information is retained, and this volume can be stored, reanalyzed, or sent online for a second opinion. The method involves relatively undemanding resources, i.e., hardware, software, competences, and time. The procedure could also benefit from refinements used in other imaging techniques to limit human-computer interactions, such as sections distortion.

EDEM3 recognizes and directs misfolded proteins to the ER-associated protein degradation (ERAD) process. EDEM3 was predicted to act as lectin or as a mannosidase because of its homology with the GH47 catalytic domain of the Man1B1, but the contribution of the other regions remained unresolved. Here, we dissect the molecular determinants governing EDEM3 function and its cellular interactions. LC/MS analysis indicates very few stable ER interactors, suggesting EDEM3 availability for transient substrate interactions. Sequence analysis reveals that EDEM3 consists of four consecutive modules defined as GH47, intermediate (IMD), protease-associated (PA), and intrinsically disordered (IDD) domain. Using an EDEM3 knock-out cell line, we expressed EDEM3 and domain deletion mutants to address EDEM3 function. We find that the mannosidase domain provides substrate binding even in the absence of mannose trimming and requires the IMD domain for folding. The PA and IDD domains deletions do not impair the trimming, but specifically modulate the turnover of two misfolded proteins, NHK and the soluble tyrosinase mutant. Hence, we demonstrate that EDEM3 provides a unique ERAD timing to misfolded glycoproteins, not only by its mannose trimming activity, but also by the positive and negative feedback modulated by the protease-associated and intrinsically disordered domain, respectively.

Various pathologies result from disruptions to or stress of endoplasmic reticulum (ER) homeostasis, such as Parkinson's disease and most neurodegenerative illnesses, diabetes, pulmonary fibrosis, viral infections, and cancers. A critical process in maintaining ER homeostasis is the selection of misfolded proteins by the ER quality-control system for destruction via ER-associated degradation (ERAD). One key protein proposed to act during the first steps of misfolded glycoprotein degradation is the ER degradation–enhancing α-mannosidase-like protein 2 (EDEM2). Therefore, characterization of the EDEM2-associated proteome is of great interest. We took advantage of using melanoma cells overexpressing EDEM2 as a cancer model system, to start documenting at the deglycoproteome level (N-glycosites identification) the emerging link between ER homeostasis and cancer progression. The dataset created for identifying the EDEM2 glyco clients carrying high mannose/hybrid N-glycans provides a comprehensive N-glycosite analysis mapping over 1000 N-glycosites on more than 600 melanoma glycoproteins. To identify EDEM2-associated proteins, we used affinity proteomics and proteome-wide analysis of sucrose density fractionation in an integrative workflow. Using intensity and spectral count–based quantification, we identify seven new EDEM2 partners, all of which are involved in ER quality-control system and ERAD. Moreover, we defined novel endogenous candidates for EDEM2-dependent ERAD by combining deglycoproteomics, stable isotope labeling with amino acids in cell culture–based proteomics, and biochemical methods. These included tumor antigens and several ER-transiting endogenous melanoma proteins, including integrin alpha-1 and protocadherin 2, the expression of which was negatively correlated with that of EDEM2. Tumor antigens are key in the antigen presentation process, whereas integrin alpha-1 and protocadherin 2 are involved in melanoma metastasis and invasion. EDEM2 could therefore have a regulatory role in melanoma through the modulation of degradation and trafficking in these glycoproteins. The data presented herein suggest that EDEM2 is involved in ER homeostasis to a greater extent than previously suggested., Graphical Abstract, Highlights • EDEM2 affinity proteomics and proteome-wide analysis of sucrose density fractionation. • New EDEM2 partners involved in ER quality control and ERAD. • Soluble tyrosinase degradation is EDEM2 dependent in melanoma cells. • Glycoproteomics and SILAC-based proteomics reveal novel EDEM2 potential substrates., In Brief Various pathologies including neurodegenerative diseases and cancers result from disruptions to or stress of ER homeostasis. One key protein proposed to act in quality control processes maintaining ER homeostasis is EDEM2. Using affinity proteomics and sucrose-density sedimentation, we identified several new EDEM2 partners involved in quality control. Moreover, we defined novel endogenous candidates for EDEM2-dependent degradation by combining glycoproteomics and SILAC-based proteomics. Our data suggest that EDEM2 is involved in ER homeostasis to a greater extent than previously thought.

Objectives To explore resident learning in the context of emergency situations. The guiding research questions were: How do residents learn in emergency situations? What factors facilitate or hinder their learning? Design A qualitative approach was used in order to understand the different perspectives of participants and explore the context of emergency situations. Aware of the complex sociocultural interactions in emergency situations, we used the methodology of constructivist grounded theory. Setting A medium-sized pediatric academic hospital in San Antonio, TX. Subjects Twenty pediatric residents participated in semi-structured interviews. Interventions None. Measurements and main results We used an iterative process of data collection and analysis. The process continued until thematic saturation was reached. The data were coded and analyzed using constant comparison. Codes were clustered into themes informed by the theory of situated learning. Several methods were used to ensure trustworthiness of results. Three main themes were identified. First, resident participation in emergency situations takes different forms. Residents participate mostly in helping roles. Watching is an under-recognized form of participation yet offered unique experiences. Managing roles are rarely afforded to residents. Second, resident participation is informed by the context of the emergency situation. Based on contextual clues (e.g., safety, needs), residents infer potential risks and values from their participation, which guides their participatory role. Residents may shift their form of participation, based on changes in the context. Supervisors play a significant role in guiding resident participation. Third, engagement, a state of cognitive and emotional focus, is critical in learning. The context of the emergency situation plays a role in the level of resident engagement. Supervisors may also foster resident engagement. Conclusions Resident participation in emergency situations is complex and informed by the context. Learning in emergency situations is influenced by residents' level of mental engagement. Supervisors may positively guide resident participation, foster engagement, and enable them to reach their learning goals in emergency situations.

Endoplasmic reticulum (ER)-associated degradation (ERAD) is the main mechanism of targeting ER proteins for degradation to maintain homeostasis, and perturbations of ERAD lead to pathological conditions. ER-degradation enhancing &alpha, mannosidase-like (EDEM1) was proposed to extract terminally misfolded proteins from the calnexin folding cycle and target them for degradation by ERAD. Here, using mass-spectrometry and biochemical methods, we show that EDEM1 is found in auto-regulatory complexes with ERAD components. Moreover, the N-terminal disordered region of EDEM1 mediates protein&ndash, protein interaction with misfolded proteins, whilst the absence of this domain significantly impairs their degradation. We also determined that overexpression of EDEM1 can induce degradation, even when proteasomal activity is severely impaired, by promoting the formation of aggregates, which can be further degraded by autophagy. Therefore, we propose that EDEM1 maintains ER homeostasis and mediates ERAD client degradation via autophagy when either dislocation or proteasomal degradation are impaired.

Background Cardiac allografts undergo characteristic alterations of the extracellular matrix, including myocardial fibrosis, that contribute to functional changes, particularly diastolic dysfunction due to increased myocardial stiffness(MS). Histological examination is the gold standard for myocardial fibrosis quantification, however, it requires endomyocardial biopsies which are invasive and not without risk. Increased native T1 and extracellular volume(ECV) using CMR T1 mapping have shown good correlation with biopsy evidence of myocardial interstitial fibrosis in heart transplant(HTx) recipients. Echocardiographic shear wave(SW) elastography is an emerging approach for measuring MS in vivo. SWs occur after mechanical excitation of the myocardium, e.g. after mitral(MVC) and aortic valve closure(AVC), and their propagation velocity is directly related to MS, thus providing an opportunity to assess stiffness at end-diastole(ED) and end-systole(ES). Purpose The aim was to investigate if natural shear wave velocities increase with the degree of diffuse myocardial fibrosis in HTx recipients. Methods We prospectively enrolled 22 HTx patients (8.8 ± 5.9 years post-HTx) that underwent CMR during their annual check-up. We performed SW elastography in parasternal long axis views of the left ventricle(LV) using an experimental scanner (HD-PULSE) equipped with a clinical phased array transducer (Samsung Medison P2-5AC) at 1100 ± 250 frames per second. Tissue acceleration maps were extracted from an anatomical M-mode line along the midline of the LV septum. The SW propagation velocity at MVC and AVC was measured as the slope on the M-mode acceleration map(FigureA). All patients underwent right heart catheterization on the same day for the measurement of pulmonary capillary wedge pressure(PCWP), as surrogate for LV filling pressure. The CMR protocol consisted of standard sequences including native and post-contrast T1 mapping. To evaluate diffuse myocardial fibrosis, native T1 and ECV were measured in the anteroseptal wall over all available short-axis slices. Results We found good correlations between SW velocities at ED and both myocardial T1 (r = 0.8,p Conclusions Both end-diastolic and end-systolic shear wave velocities showed a good correlation with CMR defined myocardial fibrosis in cardiac transplant patients. Shear wave velocities at end-diastole correlated with invasively-determined left ventricular filling pressure, reflecting the impact of the fibrous changes on the left ventricular diastolic function. These results suggest the potential of cardiac shear wave elastography for the assessment of structural changes in cardiac transplant recipients. Abstract 556 Figure.

Background The modern management of phenylketonuria (PKU) consists of generalized newborn screening (NBS) for hyperphenylalaninemia (HPA), confirmation of HPA in children detected in the NBS, introduction of dietary treatment in the first weeks of life, followed by monitoring the treatment of PKU for decades to maintain phenylalaninemia within the limits that will not affect the brain. The present study aimed to evaluate the usefulness of two chromatographic methodologies for determination of plasma Phe level in the routine management of PKU: the two dimensional thin layer chromatography (2D - TLC) and the high performance liquid chromatography (HPLC) procedures, respectively. Material and methods Samples of blood from 23 children with HPA detected by neonatal screening or with confirmed PKU who received treatment by low-Phe diet were analyzed to estimate the plasma Phe level by the two chromatographic procedures. Results In case of three subjects the very low concentrations of plasma Phe could not be detected by the 2D - TLC methodology, since the spot was not visible on the chromatogram. In four patients the differences between the values of plasma Phe determined by the two methodologies are not statistically significant, while in fifteen subjects the differences are highly statistically significant. This is due to the greater errors that appear in the case of 2D - TLC methodology. In the range of concentrations of plasma Phe higher than 360 μmol/L (which is the cut-off value for HPA), although in four cases there were statistically significant differences in the level of plasma Phe determined by the two methodologies, the value obtained by the 2D - TLC methodology was high enough to influence the decision of changing the diet so that HPA is kept under control. In addition, the intense spot of Phe on the 2D - TLC chromatogram may be detected even by un unexperienced laboratory specialist. Conclusion The HPLC procedure for measurement of plasma Phe level is very suitable to be used in the routine management of PKU. The 2D - TLC procedure may be accompanied by relatively high errors; however, it detects patients with severe PKU.

IntroductionSleep disturbances can occur as a result of major stressful events. Additionally, research evidence suggests that COVID-19 pandemic may have negatively impacted the quality of sleep among various populations. However, individuals respond differently to the stress, uncertainty and social isolation related with COVID-19 pandemic.ObjectivesThis study aimed to explore the changes in sleep quality and pattern among voluntary psychiatric patients visiting our clinic in Romania during COVID-19 pandemic.MethodsWe implemented a cross-sectional study over a period of 3 months, utilizing a Romanian-translated version of the Pittsburgh Sleep Quality Index (PSQI) which was administered through Google Forms web application. Participants lacking digital skills were provided with guidance for completing the questionnaire. Informed consent was obtained prior to participating in this study and data anonymity and confidentiality were ensured.ResultsAmong a total of 98 responders, 63% reported a global PSQI score greater than 5, indicating poor sleep. Approximately 25% of participants subjectively marked their sleep as either fairly bad or very bad. When analysing the 7 components of PSQI, our participants struggled most with long sleep latency. About a third of participants reported using sleep medication (both prescription and over-the-counter) three or more times a week within the past month.ConclusionsConsidering the fact that the current situation is likely to evolve for an unknown period of time, there is a dire need to assess the effect of prolonged adjustments in daily routine and their impact on the sleep and the quality of life of our patients.

This paper presents an algorithm for flexible and fast leader election in distributed systems using Apache Zookeeper for configuration management. The algorithm proposed in this paper is designed for applications that do not use symmetric nodes so they need a specialized election process or for applications that require a more flexible approach in the leader election process. The algorithm proposes a different approach as it allows assigning prioritizations for servers in the cluster that are candidates to become a leader. The algorithm is flexible as it takes into consideration during the leader election process of the different server settings and roles, network properties, communication latency or specific application requirements.

Economic sanctions have the reputation of harming the population in the sanctioned countries. In this paper, I investigate under what circumstances such harm occurs and what is its magnitude. I estimate the impact of in utero exposure to sanctions episodes on infant weight, child height, and child mortality. Using a large child level data set from 69 countries, I find that being exposed to sanctions leads to lower infant weight and higher probability of death before age three. I find negative effects only for children exposed to the first two years of sanctions.

An understanding of the fate of organic compounds originating from plants in soil is crucial for determining their persistence and concentrations in the environment. Aristolochic acids are believed to be the causal agents that induce Balkan endemic nephropathy by food contamination through soil adsorption of humic acids, major components of soil. Aristolochic acids are active chemicals in Aristolochia plant species found in endemic villages. In this article, molecular structure interactions between 18 structures of aristolochic acids with an inserted humic acid structure were studied. These structures were optimized in vacuo and by periodic box simulation with water solvate using the computational molecular mechanics MM+ method with HyperChem software. The QSPR models were used for correlation of the relationship between the hydrophobicity values of 18 AA structures coupled with a HA structure by MM+ and QSAR+ properties. Computational hydrophobicity values were considered dependent variables and were related to the structural features obtained by molecular and quantum mechanics calculations by multiple linear regression approaches. The obtained model was validated, and the results indicated differing hydrophobicity between the MM+ and QSAR+ properties.

Endoplasmic reticulum (ER) resident and secretory proteins that fail to reach their native conformation are selected for degradation through the ER-Associated Degradation (ERAD) pathway. The ER degradation-enhancing alpha-mannosidase-like proteins (EDEMs) were shown to be involved in this pathway but their precise role is still under investigation. Mass spectrometry analysis has contributed significantly to the characterization of protein complexes in the last years. The recent advancements in instrumentation, especially within resolution and speed can provide unique insights concerning the molecular architecture of protein-protein interactions in systems biology. Previous reports have suggested that several protein complexes in ERAD are sensitive to the extraction conditions. Indeed, whilst EDEM proteins can be recovered in most detergents, some of their partners are not solubilized, which further emphasizes the importance of the experimental setup. Here, we define such dynamic interactions of EDEM proteins by employing offline protein fractionation, nanoLC-MS/MS and describe how mass spectrometry can contribute to the characterization of such complexes, particularly within a disease context like melanoma.

The present study attempted to evaluate the carcinogenicity of natural phenolic compounds with previously demonstrated antifungal activity, using a computational structure–cytotoxicity approach, namely the quantum structure cytotoxicity relationship model. The cytotoxicity of 15 phenolic compounds with antiviral activity 96 h after treatment was studied using the AdmetSAR computational program. Per the EPA classification, four of the investigated compounds would be included in the second cytotoxicity category, four in the third category, and six showed no toxicity, rendering the studied natural phenolic compounds much less toxic to aquatic life than synthetic pesticides, the organophosphorus compounds, which mostly fall into the first and second categories of toxicity.

The implementation of a fault-tolerant system requires some type of consensus algorithm for correct operation. From Paxos to View-stamped Replication and Raft multiple algorithms have been developed to handle this problem. This paper presents and compares the Raft algorithm and Apache Kafka, a distributed messaging system which, although at a higher level, implements many concepts present in Raft (strong leadership, append-only log, log compaction, etc.).This shows that mechanisms conceived to handle one class of problems (consensus algorithms) are very useful to handle a larger category in the context of distributed systems.

The degradation process of the antigens specific to MHC-I presentation depends mainly on the proteasomal proteases in the cytosol. However, since many antigens are glycoproteins, including tumor antigens or viruses envelope proteins, their glycosylation status could also affect their processing and presentation. Here, we investigate the processing of tyrosinase, a multiple glycosylated tumor antigen overexpressed in human malignant melanoma. By LC-MS/MS analysis of human tyrosinase expressed in a melanoma cell, we show that all seven sites of tyrosinase are at least partially N-glycosylated. Using human CD8+ T-cell clones specific for the tyrosinase epitope YMDGTMSQV (369-377), including an N-glycosylation site, we found that transfectants of single and triple N-glycosylation mutants are recognized by specific T cells. Importantly, single, triple, and the aglycosylated tyrosinase mutants lacking the epitope located N-glycosylation site (N371D) were able to trigger higher CD8+ T-cell activation. The LC/MS analysis showed significant increase of the amount of YMDGTMSQV peptide resulted from accelerated oligomerization and degradation of aglycosylated mutants. The generation of the antigenic peptide by the antigen processing machinery is therefore largely independent of tyrosinase N-glycosylation. However, while distal N-glycans had no effect on the epitope generation, the mutants lacking the N371 glycan generated the antigenic peptide more efficiently. We conclude that epitope located N-glycans limit the ability of human tyrosinase to provide HLA-A2-restricted antigen for recognition by specific CD8+ T cells.

Eurypanopeus depressus (Smith, 1869) was observed in Constanţa on the Romanian Black Sea shore in 2013. Herein, this species, which was previously only reported from the western Atlantic, is recorded for the first time in European waters. Some morphological details significant for the species are described, with an emphasis on the setae of the 3rd maxilliped.

Chronic Obstructive Pulmonary Disease (COPD) is a clinical syndrome characterised by a slow progressive decline in expiratory airflow [1], a process that has gradually developed over the years. Studies of patients with COPD show an inflammatory process in the small airways [2]. The aim of this paper is to identify the cytopathological aspects of the liquids present in the bronchoalveolar lavage in patients with COPD. We were performed a descriptive analytical case-control and prospective study on forty patients with COPD and ten asymptomatic smokers (healthy smokers or patients at risk). The percentage of marcophage, the type of the dominant inflamatory cell, in the bronchoalveolar lavage (BAL) liquid was significantly higher at patients with mild and moderate COPD as compared to patients with severe and very severe COPD. In the present work, the percentage of the neutrophil in the BAL liquid was significantly higher at patients with severe and very severe COPD, as compared to the patients with mild and moderate COPD and to aparently healthy smokers. In conclusion, we can say that COPD is characterized by an inflammatory process located in the small airways with predominant participation of macrophages, the procentage of macrophages in BAL fluid variyng inversely proportional to the severity of the disease.

Transcription machinery plays a central role in both the gene expression and nucleoid compaction. In this chapter we elaborate on the optimization of RNA polymerase purification protocol using a mild procedure with the purpose of preserving its native composition. This protocol combines protein extraction under non-denaturing conditions, heparin based affinity purification, and consequent BN-PAGE-SDS-PAGE separation. The outcome is an experimental procedure for screening RNA polymerase composition with associated proteins, in various bacterial strains or mutant backgrounds. With modifications in the column purification step, this procedure can be applied for isolation and identification of the components of other multi-protein complexes.

In the last two decades, there has been registered a constant decrease in the number of breastfed infants and in the duration of breastfeeding. Our paper is aiming at pointing out the rate of breastfeeding in a group of infants correlated with mothers’ demographic profile, the ways to give birth, the time until the initiation of breastfeeding, breastfeeding counselling, and the mothers’ educational level. The study had a prospective development, analysing the answers provided by the mothers of breastfed infants, who filled in some anonymous questionnaires given by family doctors and the 2 nd Paediatric Clinic of the Emergency County Hospital Craiova. The study comprised 110 mothers, with an average age of 20–25 years, 69% of them coming from an urban environment. 56% of the cases were first born children (only 9% being the third born child or more). In 60% of the cases, children were born by caesarian section. 42% of the mothers had a higher education level, 29% having highschool education, while the rest of them having primary and secondary school education. The moment they filled in the questionnaires, 47.3% of the mothers were breastfeeding their infants of 4 months of age, on average. 49% of the infants had an early breastfeeding, while 60% of the mothers needed counselling offered by nurses or neonatologists. 14.5% of the mothers declared they were never explained the breastfeeding technique. Nearly 75% of the mothers were never monitored during their breastfeeding process, at home. Birth by caesarian section, lack of breastfeeding monitoring, and mothers’ lack of education may be the causes of the low rate of breastfeeding in infants.

Transcription machinery plays a central role in both the gene expression and nucleoid compaction. In this chapter we elaborate on the optimization of RNA polymerase purification protocol using a mild procedure with the purpose of preserving its native composition. This protocol combines protein extraction under non-denaturing conditions, heparin based affinity purification, and consequent BN-PAGE-SDS-PAGE separation. The outcome is an experimental procedure for screening RNA polymerase composition with associated proteins, in various bacterial strains or mutant backgrounds. With modifications in the column purification step, this procedure can be applied for isolation and identification of the components of other multi-protein complexes.

Dopachrome tautomerase (DCT) and tyrosinase (Tyr) are melanogenic enzymes and structurally related melanosomal proteins. The present study investigates DCT expression comparatively with Tyr, the most tested melanoma biomarker, aiming to evaluate DCT potential in the assessment of melanocytic tumors and gain insights into the molecular and pathological characterization of DCT-phenotype in tumor progression. DCT and Tyr are simultaneously analyzed in melanoma cell lines by semiquantitative RT-PCR, western blot, and N-glycan analysis, and in cell populations of melanocytic tumors by immunohistofluorescence using a novel anti-hDCT antibody against an extended sequence within DCT luminal domain. DCT, unlike Tyr, is fully processed along the secretory pathway in both pigmented and amelanotic melanoma cells. In 53 nevi and 116 primary malignant melanomas, 81% and 52%, respectively, are DCT+/Tyr+, showing that DCT is a stable antigen, retained by most tumors and partially expressed in Tyr-negative cell populations. The DCT/Tyr disjunction is a process correlated with melanocyte neoplastic transformation and malignant progression. A tumor architecture--DCT-phenotype-containing DCT+/Tyr- cell populations selected into the innermost dermis from double-positive cells is detected in 35% of DCT+/Tyr+ specimens. The DCT-phenotype is associated with enhanced neurotization in benign nevi and with ulceration in thin malignant melanomas. The intradermal DCT+/Tyr- clones in superficial melanomas acquire the expression and specific subcellular distribution of unfavorable prognostic markers. DCT assessment shows specific antigen patterns with potential significance in the outcome of melanocytic lesions, connecting DCT, a mediator of a melanoma stress-resistant pathway, and an antiapoptotic molecule to DCT- phenotypes that are possibly more stable and stress resistant.

The electronic connections need to satisfy three functionalities and those are the electrical, thermal and mechanical functionalities. The PCB (Printed Circuit Board) has the role of providing a mechanical support for an assembly of various electrical and electronical components and to connect them to obtain a functional end-product. The goal of the present paper was to identify the mechanical and tribological properties of the lead-free soldering alloy SAC 305 deposit on the copper-PCB assembly used in electronic technology. The variation of the friction coefficient for all the normal forces applied on the test samples was analyzed. The understanding of the behavior of the lead-free soldering alloy for the micro scratch test is essential for the improvement of the hole electronic system integrity.