Miracle-dz/UniFAR-FTUs huggingface dataset:
Translocation and Endocytosis for Cell-penetrating Peptide Internalization

Cell-penetrating peptides (CPPs) share the property of cellular internalization. The question of how these peptides reach the cytoplasm of cells is still widely debated. Herein, we have used a mass spectrometry-based method that enables quantification of internalized and membrane-bound peptides. Internalization of the most used CPP was studied at 37 °C (endocytosis and translocation) and 4 °C (translocation) in wild type and proteoglycan-deficient Chinese hamster ovary cells. Both translocation and endocytosis are internalization pathways used by CPP. The choice of one pathway versus the other depends on the peptide sequence (not the number of positive changes), the extracellular peptide concentration, and the membrane components. There is no relationship between the high affinity of these peptides for the cell membrane and their internalization efficacy. Translocation occurs at low extracellular peptide concentration, whereas endocytosis, a saturable and cooperative phenomenon, is activated at higher concentrations. Translocation operates in a narrow time window, which implies a specific lipid/peptide co-import in cells.
Thrombin generation and bleeding in haemophilia A

Summary. Haemophilia A displays phenotypic heterogeneity with respect to clinical severity. The aim of this study was to determine if tissue factor (TF)‐initiated thrombin generation profiles in whole blood in the presence of corn trypsin inhibitor (CTI) are predictive of bleeding risk in haemophilia A. We studied factor(F) VIII deficient individuals (11 mild, 4 moderate and 12 severe) with a well‐characterized 5‐year bleeding history that included haemarthrosis, soft tissue haematoma and annual FVIII concentrate usage. This clinical information was used to generate a bleeding score. The bleeding scores (range 0–32) were separated into three groups (bleeding score groupings: 0, 0 and ≤9.6, >9.6), with the higher bleeding tendency having a higher score. Whole blood collected by phlebotomy and contact pathway suppressed by 100 μg mL−1 CTI was stimulated to react by the addition of 5 pm TF. Reactions were quenched at 20 min by inhibitors. Thrombin generation, determined by enzyme‐linked immunosorbent assay for thrombin–antithrombin was evaluated in terms of clot time (CT), maximum level (MaxL) and maximum rate (MaxR) and compared to the bleeding score. Data are shown as the mean±SD. MaxL was significantly different (P < 0.001) between the groups: 504 ± 114, 315 ± 117 and 194 ± 91 nm; with higher thrombin concentrations in the groups with lower bleeding scores. MaxR was higher in the groups with a lower bleeding score; 97 ± 51, 86 ± 60 and 39 ± 16 nm min−1 (P = 0.09). No significant difference was detected in CT among the groups, 5.6 ± 1.3, 4.7 ± 0.7 and 5.6 ± 1.3 min. Our empirical study in CTI‐inhibited whole blood shows that the MaxL of thrombin generation appears to correlate with the bleeding phenotype of haemophilia A.
Methadone maintenance treatment in China: perceived challenges from the perspectives of service providers and patients.

BACKGROUND China has recently adopted methadone maintenance treatment (MMT) as a national strategy to address the problem of drug abuse and related public health issues such as HIV and HCV infections. However, low enrollment and retention rates suggest that barriers may exist in MMT utilization. This study examined both patients' perceptions and service providers' perceptions of challenges in MMT implementation in China. METHODS Four focus groups were conducted in two Chinese cities, Shanghai and Kunming, to explore the perceived and experienced barriers in MMT participation in China. All focus group discussions with participants were audio taped and transcribed. Atlas.ti 5.1 was used to analyze data. RESULTS Service providers and patient participants reported positive experiences (e.g. effects of MMT in curbing withdrawal symptoms) but also expressed concerns about side effects and continued heroin use during MMT. They also identified barriers in participating and remaining in MMT, including affordability (fee requirement), acceptability (methadone as a substitution, dose, long-term nature), accommodation and accessibility (inconvenient operation hours, lack of transferability to other MMT clinics during travel) and competition between public health and public security. CONCLUSIONS The present findings have implications for reconsidering the current MMT policies and practices in order to improve access, utilization and, ultimately, the effectiveness of MMT in China.
Factors associated with bronchiolitis obliterans syndrome and chronic graft‐versus‐host disease after allogeneic hematopoietic cell transplantation

Bronchiolitis obliterans syndrome (BOS) is a form of chronic graft vs. host disease (cGVHD) and a highly morbid pulmonary complication after allogeneic hematopoietic stem cell transplantation (HSCT). We assessed the prevalence and risk factors for BOS and cGVHD in a cohort of HSCT recipients, including those who received reduced intensity conditioning (RIC) HSCT. Between January 1, 2000 and June 30, 2010, all patients who underwent allogeneic HSCT at our institution (n = 1854) were retrospectively screened for the development of BOS by PFT criteria. We matched the BOS cases with two groups of control patients: (1) patients who had concurrent cGVHD without BOS and (2) those who developed neither cGVHD nor BOS. Comparisons between BOS patients and controls were conducted using t‐test or Fisher's exact tests. Multivariate regression analysis was performed to examine factors associated with BOS diagnosis. All statistical analyses were performed using SAS 9.2. We identified 89 patients (4.8%) meeting diagnostic criteria for BOS at a median time of 491 days (range: 48–2067) after HSCT. Eighty‐six (97%) of our BOS cohort had extra‐pulmonary cGVHD. In multivariate analysis compared to patients without cGVHD, patients who received busulfan‐based conditioning, had unrelated donors, and had female donors were significantly more likely to develop BOS, while ATG administration was associated with a lower risk of BOS. Our novel results suggest that busulfan conditioning, even in RIC transplantation, could be an important risk factor for BOS and cGVHD. Am. J. Hematol. 89:404–409, 2014. © 2013 Wiley Periodicals, Inc.
Mucocele labial: relato de caso em criançade dois anos de idade

Mucoceles are common lesions of the oral mucosa that result when a salivary gland duct ruptures. These ducts are subject to trauma and consequent laceration, with mucus extruding into the surrounding soft tissues. The lesion is seen mainly among children and young adults and has no predilection based on gender. It can develop in any location harboring a minor salivary gland. This article presents a case to mucocele in a child of 2 years of age, of traumatic origin. The clinical boarding of the case, and the treatment, will be told and argued.
Importance of Membrane Structural Integrity for RPE65 Retinoid Isomerization Activity*

Regeneration of visual chromophore in the vertebrate visual cycle involves the retinal pigment epithelium-specific protein RPE65, the key enzyme catalyzing the cleavage and isomerization of all-trans-retinyl fatty acid esters to 11-cis-retinol. Although RPE65 has no predicted membrane spanning domains, this protein predominantly associates with microsomal fractions isolated from bovine retinal pigment epithelium (RPE). We have re-examined the nature of RPE65 interactions with native microsomal membranes by using extraction and phase separation experiments. We observe that hydrophobic interactions are the dominant forces that promote RPE65 association with these membranes. These results are consistent with the crystallographic model of RPE65, which features a large lipophilic surface that surrounds the entrance to the catalytic site of this enzyme and likely interacts with the hydrophobic core of the endoplasmic reticulum membrane. Moreover, we report a critical role for phospholipid membranes in preserving the retinoid isomerization activity and physical properties of RPE65. Isomerase activity measured in bovine RPE was highly sensitive to phospholipase A2 treatment, but the observed decline in 11-cis-retinol production did not directly reflect inhibition by products of lipid hydrolysis. Instead, a direct correlation between the kinetics of phospholipid hydrolysis and retinoid isomerization suggests that the lipid membrane structure is critical for RPE65 enzymatic activity. We also provide evidence that RPE65 operates in a multiprotein complex with retinol dehydrogenase 5 and retinal G protein-coupled receptor in RPE microsomes. Modifications in the phospholipid environment affecting interactions with these protein components may be responsible for the alterations in retinoid metabolism observed in phospholipid-depleted RPE microsomes. Thus, our results indicate that the enzymatic activity of native RPE65 strongly depends on its membrane binding and phospholipid environment.
Bone marrow 18F‐fluoro‐2‐deoxy‐d‐glucose positron emission tomography/computed tomography cannot replace bone marrow biopsy in diffuse large B‐cell lymphoma

This study aimed to investigate whether visual and quantitative 18F‐fluoro‐2‐deoxy‐d‐glucose positron emission tomography/computed tomography (FDG‐PET/CT)‐based bone marrow assessment can replace blind bone marrow biopsy (BMB) in newly diagnosed diffuse large B‐cell lymphoma (DLBCL). This retrospective study included 78 patients with newly diagnosed DLBCL who had undergone both FDG‐PET/CT and BMB. FDG‐PET/CT images were visually evaluated for bone marrow involvement. Patient‐based sensitivity of visual FDG‐PET/CT assessment was calculated using BMB as the reference standard. Metabolically active volume, maximum standardized uptake value, 3D partial volume corrected mean standardized uptake value, and 3D partial volume corrected mean metabolic volume product (cMVPmean) of FDG‐avid bone marrow lesions were measured. Cox regression analysis was used to determine the influence of (potential) prognostic factors (BMB status, visual [dichotomous] FDG‐PET/CT bone marrow status, metabolically active volume, maximum standardized uptake value, 3D partial volume corrected mean standardized uptake value, 3D partial volume corrected mean metabolic volume product, and International Prognostic Index score) on progression‐free survival and overall survival. FDG‐PET/CT detected bone marrow involvement in 34 (43.6%) cases and BMB in 16 (20.5%) of 78 cases, of whom 11 were also detected by FDG‐PET/CT, resulting in a patient‐based sensitivity of 68.8% (95% confidence interval = 44.2%–86.1%) for FDG‐PET/CT. In the multivariate Cox proportional hazards model, only BMB status was an independent predictive factor of progression‐free survival (P = 0.016) and overall survival (P = 0.004). In conclusion, FDG‐PET/CT misses bone marrow involvement that has been detected by BMB in a non‐negligible proportion of patients. Furthermore, both visual and quantitative FDG‐PET/CT‐based bone marrow assessments are prognostically inferior to BMB. Therefore, FDG‐PET/CT cannot replace BMB in newly diagnosed DLBCL. Am. J. Hematol. 89:726–731, 2014. © 2014 Wiley Periodicals, Inc.
BclxL Changes Conformation upon Binding to Wild-type but Not Mutant p53 DNA Binding Domain*

p53 can induce apoptosis through mitochondrial membrane permeabilization by interaction of its DNA binding region with the anti-apoptotic proteins BclxL and Bcl2. However, little is known about the action of p53 at the mitochondria in molecular detail. By using NMR spectroscopy and fluorescence polarization we characterized the binding of wild-type and mutant p53 DNA binding domains to BclxL and show that the wild-type p53 DNA binding domain leads to structural changes in the BH3 binding region of BclxL, whereas mutants fail to induce such effects due to reduced affinity. This was probed by induced chemical shift and residual dipolar coupling data. These data imply that p53 partly achieves its pro-apoptotic function at the mitochondria by facilitating interaction between BclxL and BH3-only proteins in an allosteric mode of action. Furthermore, we characterize for the first time the binding behavior of Pifithrin-μ, a specific small molecule inhibitor of the p53-BclxL interaction, and present a structural model of the protein-ligand complex. A rather unusual behavior is revealed whereby Pifithrin-μ binds to both sides of the protein-protein complex. These data should facilitate the rational design of more potent specific BclxL-p53 inhibitors.
A Novel Glucosyltransferase Is Required for Glycosylation of a Serine-rich Adhesin and Biofilm Formation by Streptococcus parasanguinis*

Fap1-like serine-rich glycoproteins are conserved in streptococci, staphylococci, and lactobacilli, and are required for bacterial biofilm formation and pathogenesis. Glycosylation of Fap1 is mediated by a gene cluster flanking the fap1 locus. The key enzymes responsible for the first step of Fap1 glycosylation are glycosyltransferases Gtf1 and Gtf2. They form a functional enzyme complex that catalyzes the transfer of N-acetylglucosamine (GlcNAc) residues to the Fap1 polypeptide. However, until now nothing was known about the subsequent step in Fap1 glycosylation. Here, we show that the second step in Fap1 glycosylation is catalyzed by nucleotide-sugar synthetase-like (Nss) protein. The nss gene located upstream of fap1 is also highly conserved in streptococci and lactobacilli. Nss-deficient mutants failed to catalyze the second step of Fap1 glycosylation in vivo in Streptococcus parasanguinis and in a recombinant Fap1 glycosylation system. Nss catalyzed the direct transfer of the glucosyl residue to the GlcNAc-modified Fap1 substrate in vitro, demonstrating that Nss is a glucosyltransferase. Thus we renamed Nss as glucosyltransferase 3 (Gtf3). A gtf3 mutant exhibited a biofilm defect. Taken together, we conclude that this new glucosyltransferase mediates the second step of Fap1 glycosylation and is required for biofilm formation.
Helpful advice and hidden expertize: pharmacy users' experiences of community pharmacy accessibility

Background In recent years community pharmacies have emerged as strategically important settings to deliver services aimed at promoting public health. In order to develop evidence‐based approaches to public health interventions that exploit the unique accessibility of community pharmacies, it is important to determine how people experience care in this context. This study, therefore, aimed to describe how care is perceived and experienced in community pharmacies with particular focus on community pharmacy access. Methods In‐depth semi‐structured interviews were used to explore the perceptions and experiences' of people using community pharmacies. Results A total of 30 participants were interviewed. Themes specifically emerged in relation to community pharmacy access; these fell into four main categories: relationships; time; lack of awareness; and empowerment. Conclusions The experience of developing a trusting relationship with the pharmacist is an important consideration in the context of community pharmacy accessibility. This could be an important consideration when a person uses a community pharmacy to access a public health service. There is also a perceived lack of awareness among the general public about the extended role of community pharmacy; this is a potential barrier toward people using them.
Neural-specific Deletion of FIP200 Leads to Cerebellar Degeneration Caused by Increased Neuronal Death and Axon Degeneration*

FIP200 (FAK family-interacting protein of 200 kDa) is a conserved protein recently identified as a potential mammalian counterpart of yeast autophagy protein Atg17. However, it remains unknown whether mammalian FIP200 regulates autophagy in vivo. Here we show that neural-specific deletion of FIP200 resulted in cerebellar degeneration accompanied by progressive neuronal loss, spongiosis, and neurite degeneration in the cerebellum. Furthermore, deletion of FIP200 led to increased apoptosis in cerebellum as well as accumulation of ubiquitinated protein aggregates without any deficiency in proteasome catalytic functions. We also observed an increased p62/SQSTM1 accumulation in the cerebellum and reduced autophagosome formation as well as accumulation of damaged mitochondria in the mutant mice. Lastly, analysis of cerebellar neurons in vitro showed reduced JNK activation and increased susceptibility to serum deprivation-induced apoptosis in cerebellar neurons from the mutant mice. Taken together, these results provide strong genetic evidence for a role of FIP200 in the regulation of neuronal homeostasis through its function in autophagy in vivo.
Consumer segmentation and time interval between types of hospital admission: a clinical linkage database study

Background Healthcare policies target unplanned hospital admissions and 30-day re-admission as key measures of efficiency, but do not focus on factors that influence trajectories of different types of admissions in the same patient over time. Objectives To investigate the influence of consumer segmentation and patient factors on the time intervals between different types of hospital admission. Research design, subjects and measures A cohort design was applied to an anonymised linkage database for adults aged 40 years and over (N = 58 857). Measures included Mosaic segmentation, multimorbidity defined on six chronic condition registers and hospital admissions over a 27-month time period. Results The shortest mean time intervals between two consecutive planned admissions were: 90 years and over (160 days (95% confidence interval (CI): 146-175)), Mosaic groups 'Twilight subsistence' (171 days (164-179)) or 'Welfare borderline' and 'Municipal dependency' (177 days (172-182)) compared to the reference Mosaic groups (186 days (180-193)), and multimorbidity count of four or more (137 days (130-145)). Mosaic group 'Twilight subsistence' (rate ratio (RR) 1.22 (95% CI: 1.08-1.36)) or 'Welfare borderline' and 'Municipal dependency' RR 1.20 (1.10-1.31) were significantly associated with higher rate to an unplanned admission following a planned event. However, associations between patient factors and unplanned admissions were diminished by adjustment for planned admissions. Conclusion Specific consumer segmentation and patient factors were associated with shorter time intervals between different types of admissions. The findings support innovation in public health approaches to prevent by a focus on long-term trajectories of hospital admissions, which include planned activity.
Nurse Manager Emotional Intelligence as a Predictor to Registered Nurse Job Satisfaction and RN Perceptions of the Practice Environment and the Relationship to Patient, Nursing and Hospital Outcomes

The purpose of this study was to determine if the level of Nurse Manager (NM) emotional intelligence (EI) predicted registered nurse (RN) job satisfaction and RN perceptions of the practice environment. In addition, relationships to patient, nursing and hospital outcomes were explored. Participants included RNs (N=659) and NMs (N=38) from 53 nursing units at eight hospitals located in the southeast region of the United States. A cross-sectional, correlational research design was used to test the hypotheses. Pearson product-moment correlation coefficients, simple linear and multiple regression statistics were conducted to analyze the data. Level of NM EI had a positive, not significant relationship to RN job satisfaction (B = 3.63, p<.373) and RN perceptions of the practice environment (B = 2.79, p<.189). A direct, positive significant relationship was observed between the variables NM EI and patient satisfaction with nursing care (B=.269, p<.001). There was a positive, significant relationship noted between the variables RN job satisfaction and RN perceptions of the practice environment (r= .762, p<.001). The indirect relationships between level of NM EI and patient, nursing, and hospital outcomes were not significant. There was a direct significant, positive relationship noted between the variables RN perceptions of the practice environment and patient satisfaction with nursing care (p<.044). In addition, the interaction between RN job satisfaction and RN hours of care had a positive, significant relationship with unit
C1272F: a novel type 2A von Willebrand’s disease mutation in A1 domain; its clinical significance

Summary. Most mutations identified in 2A VWD patients are localized in the A2 domain, although missense substitutions have also been recognized in the A1 domain. We describe a novel heterozygous missense mutation in the A1 domain of VWF gene responsible for type 2A phenotype. Analysis of the complete exon 28 was carried out in a patient and his mother with life‐long histories of moderate to severe bleeding and laboratory data of type 2A VWD. The analysis of exon 28 of VWF gene showed a 3815 G→T transversion resulting in C1272F mutation. It is probably associated with a group I mechanism according to patients’ clinical symptoms, and, in the case of the propositus, the lack of clinical response to treatment with desmopressin. The mutation was not found in 100 normal alleles. This substitution affected the normal S–S bound between C1272 and C1458, which is involved in A1 loop structure, altering the normal multimerization and function of VWF. The VWFpp/VWF:Ag ratio in the propositus and his mother was >3, suggesting a shortened survival of VWF. We believe it is important to report the complete clinical phenotype corresponding to the new mutation to increase the knowledge in the clinical field.
Variation in cancer outcomes amongst children and young adults in Yorkshire

This study set out to improve upon the quality of research addressing variation in cancer outcomes amongst children and young adults (CYAs) through the novel application of multiple imputation (MI) to the population based Yorkshire cancer register. The study also sought to determine whether there were inequalities in disease severity according to age, ethnicity or deprivation for CYAs with cancer for the first time in the UK and to quantify cardiovascular late effects amongst survivors of CYA cancer based on a unique data linkage approach to hospital admission data. Key survival inequalities for CYAs with central nervous system (CNS) tumours (n=795), leukaemia (n=912) and germ cell tumours (GCTs) (n=846) between 1990 and 2009 were identified. Teenagers and young adults (TYA) compared to children across all three groups and those of non-white and non-Asian ethnicity diagnosed with CNS tumours had significantly poorer survival. Importantly, these survival inequalities could not be explained by differences in the stage at diagnosis according to age, ethnicity or deprivation. Survival rates from CNS tumours and leukaemia continued to improve over time. These improvements only became evident after using MI to adjust for missingness, which is often ignored by researchers. Despite no observed improvement for GCTs over time, the number of advanced stage tumours at diagnosis decreased significantly for this diagnostic group. For all cancers combined, the long term cardiovascular effects of cancer exist not only for children, but also for TYAs surviving their cancer. Continued efforts should be made to ensure equal access to clinical trials and improved treatment protocols for TYAs. In addition, children as well as TYAs should be monitored for early signs of cardiovascular disease to maximise cardiovascular health. Finally, ignoring missing data can result in reduced study power and biased estimates, thus researchers should strive to use advanced techniques such as MI to account for missing data.