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Background/Objectives: Para‐phenylenediamine, a dye frequently added to henna tattoos to create the black color, is a potent contact allergen. Severe contact dermatitis may arise within days even after the first application. Our objective was to develop a method for rapid and complete removal of para‐phenylenediamine‐containing black henna tattoos from the skin, an important problem many physicians are confronted with, but for which no simple method exists. Methods: A database search revealed polyethylene glycol 400, described in removal of ortho‐phenylenediamine from contaminated skin. We therefore investigated its use in removal of the structurally related ortholog para‐phenylenediamine present in black henna tattoos. Results: A protocol was established involving repeated cycles of rinsing of the skin with polyethylene glycol 400 solution. In 5 patients, one of whom had already developed a severe blistering contact dermatitis and another a mild erosive dermatitis, black henna tattoos were successfully removed. Removal was completed in a single session of 1 hour or less, depending on tattoo size, with a maximum of 6 rinse cycles. Conclusions: We provide a simple and safe method for rapid and effective removal of black henna tattoos. This procedure requires no special equipment and can be applied in virtually any setting. [ABSTRACT FROM AUTHOR]

The aim of the study is to explore the nature of understanding when learning music through the use of 'graphic representations' trialled in a learning conversation with a ten-year old flautist. It is argued that the powerful visual component of presenting a musical score in graphic form can enhance students' understanding and ability to process the score more effectively by providing a succinct way of accessing the data. Central to understanding the analysis of sonata form movements is the need to create a representation which is independent from the existing score. This study offers a practical way of doing this which has the potential for wider application. [ABSTRACT FROM AUTHOR]

Abstract: Mast cells are important players in innate immunity and mediate allergic responses. Upon stimulation, they release biologically active mediators including histamine, cytokines and lysosomal hydrolases. We used permeabilized rat basophilic leukaemia cells as model to identify R-SNAREs (soluble NSF (N-ethylmaleimide-sensitive fusion protein)) mediating exocytosis of hexosaminidase from mast cells. Of a complete set of recombinant mammalian R-SNAREs, only vesicle associated membrane protein (VAMP8)/endobrevin consistently blocked hexosaminidase release, which was also insensitive to treatment with clostridial neurotoxins. Thus, VAMP8, which also mediates fusion of late endosomes and lysosomes, plays a major role in hexosaminidase release, strengthening the view that mast cell granules share properties of both secretory granules and lysosomes. [Copyright &y& Elsevier]

We report on the isolation, sequence and a putative role of a human endoplasmic-reticulum-lumenal protein, ERp28. The protein has the C-terminal retention signal KEEL and localizes to the endoplasmic reticulum (ER) as seen by subcellular fractionation and immunofluorescence studies. The protein has significant sequence similarity to members of the protein disulfide isomerase (PDI) family, although it lacks the thioredoxin box (CGHC) motif. We propose, on the basis of sequence analysis, a model of the domain structure of PDI, representing a significant extension of previously proposed models. Our results are in partial agreement with recently published NMR data [Kemmink, J., Darby, J., Dijkstra, K., Nilges, M. & Creighton, T. E. (1997) Curr. Biol. 7, 239-245] and indicate that PDI contains, in addition to the two thioredoxin folds described in previous models, two thioredoxin folds within the domains previously defined as b and b′. The thioredoxin domain of ERp28 shares a higher degree of similarity with the corresponding active and inactive domains of PDI than with other members of the PDI family, indicating that ERp28 developed from an ancient form of PDI or a PDI precursor. In contrast to Ig-heavy-chain-binding protein, human ERp28 is not induced by metabolic stress (tunicamycin). In in vitro experiments, ERp28 and calnexin precipitate with overexpressed, wild-type hepatitis B small surface antigen and with a mutated ER-retained form. This indicates that ERp28, as calnexin, may be involved in the processing of secretory proteins within the ER. [ABSTRACT FROM AUTHOR]

E-learning/online transfusion education A total of 12 of 14 respondents confirmed the use of e-learning platforms in their hospitals/health services for transfusion education (Table 4). The Serious Hazards of Transfusion 2021 report highlighted the importance of providing training and regular education for healthcare professionals involved in the transfusion process to achieve safe transfusion [[1]]. Transfusion education for personnel who administer transfusions The majority of the hospitals/health services require mandatory transfusion education for personnel who administer transfusions, while four hospitals/health services have optional education (Table 2). [Extracted from the article]

Section 5 The main challenge in maintaining a transfusion education programme for personnel who administer transfusions in our hospital is organizing the schedule of the staff to attend the transfusion education. Section 2: Transfusion education for personnel who administer transfusions The following types of transfusion education are available for personnel who administer transfusions in our hospital/health service: Independent e-learning. Section 3 Yes, e-learning/online transfusion education is used for personnel who administer transfusions in our hospital/health service. e-learning/online transfusion education has been used for 9 years in our hospital. Section 3 Yes, e-learning/online transfusion education is used for personnel who administer transfusions in our hospital/health service. e-learning/online education has been used for approximately 5 years in our nursing department. [Extracted from the article]

Abstract: The protein disulfide isomerase (PDI) family member ERp46/endoPDI/thioredoxin domain-containing protein 5 is preferentially expressed in a limited number of tissues, where it may function as a survival factor for nitrosative stress in vivo. It is involved in insulin production as well as in adiponectin signaling and interacts specifically with the redox-regulatory endoplasmic reticulum proteins endoplasmic oxidoreductin 1α (Ero1α) and peroxiredoxin-4. Here, we show that ERp46, although lacking a PDI-like redox-inactive b′-thioredoxin domain with its hydrophobic substrate binding site, is able to bind to a large pool of peptides containing aromatic and basic residues via all three of its catalytic domains (a0, a and a′), though the a0 domain may contain the primary binding site. ERp46, which shows relatively higher activity as a disulfide-reductase than as an oxidase/isomerase in vitro compared to PDI and ERp57, possesses chaperone activity in vivo, a property also shared by the C-terminal a′ domain. A crystal structure of the a′ domain is also presented, offering a view of possible substrate binding sites within catalytic domains of PDI proteins. [Copyright &y& Elsevier]

Abstract: The protein disulfide isomerase-related protein ERp29 is a putative chaperone involved in processing and secretion of secretory proteins. Until now, however, both the structure and the exact nature of interacting substrates remained unclear. We provide for the first time a crystal structure of human ERp29, refined to 2.9 Å, and show that the protein has considerable structural homology to its Drosophila homolog Wind. We show that ERp29 binds directly not only to thyroglobulin and thyroglobulin-derived peptides in vitro but also to the Wind client protein Pipe and Pipe-derived peptides, although it fails to process Pipe in vivo. A monomeric mutant of ERp29 and a D domain mutant in which the second peptide binding site is inactivated also bind protein substrates, indicating that the monomeric thioredoxin domain is sufficient for client protein binding. Indeed, the b domains of ERp29 or Wind, expressed alone, are sufficient for binding proteins and peptides. Interacting peptides have in common two or more aromatic residues, with stronger binding for sequences with overall basic character. Thus, the data allow a view of the two putative peptide binding sites of ERp29 and indicate that the apparent, different processing activity of the human and Drosophila proteins in vivo does not stem from differences in peptide binding properties. [Copyright &y& Elsevier]

The peptidyl prolyl cis/ trans isomerase (PPIase) Pin1 plays an important role in phosphorylation-dependent events of the cell cycle. This function is linked to its display of two phosphothreonine/phosphoserine-proline binding motifs, one within the type IV WW domain and a second within the parvulin-like catalytic domain. By microinjection of the compound Ac-Phe-D-Thr(PO3H2)-Pip-Nal-Gln-NH2, which inhibits Xenopus laevis Pin1 with a Ki value of 19.4±1.5 nM, into the animal pole of X. laevis embryos at the two-cell stage, the impact of Pin1 PPIase activity on cell cycle progression and embryonic development could be analysed, independent of WW domain-mediated phosphoprotein binding. Injected embryos showed a dramatically decreased survival rate at late stages of development that could only be partially compensated by co-injection with mRNAs of enzymatically active Pin1 variants, demonstrating that the phosphorylation-specific PPIase activity of Pin1 is essential for cell division and development in X. laevis. [ABSTRACT FROM AUTHOR]

The structure and mode of binding of the endoplasmic reticulum protein disulfide isomerase-related proteins to their substrates is currently a focus of intensive research. We have recently determined the crystal structure of the Drosophila melanogaster protein disulfide isomerase-related protein Wind and have described two essential substrate binding sites within the protein, one within the thioredoxin b-domain and another within the C-terminal D-domain. Although a mammalian ortholog of Wind (ERp29/28) is known, conflicting interpretations of its structure and putative function have been postulated. Here, we have provided evidence indicating that ERp29 is indeed similar in both structure and function to its Drosophila ortholog. Using a site-directed mutagenesis approach, we have demonstrated that homodimerization of the b-domains is significantly reduced in vitro upon replacement of key residues at the predicted dimerization interface. Investigation of Wind-ERp29 fusion constructs showed that mutants of the D-domain of ERp29 prevent transport of a substrate protein (Pipe) in a manner consistent with the presence of a discrete, conserved peptide binding site in the D-domain. Finally, we have highlighted the general applicability of these findings by showing that the D-domain of a redox-active disulfide isomerase, from the slime mold Dictyostelium discoideum, can also functionally replace the Wind D-domain in vivo. [ABSTRACT FROM AUTHOR]

The structures of the PDI-related protein Wind (with a C-­terminal His6 tag) and the mutants Y53S, Y53F and Y55K have been determined and compared with the wild-type structure with the His6 tag at the N-terminus. All five structures show the same mode of dimerization, showing that this was not an artefact introduced by the nearby N-terminal His6 tag and suggesting that this dimer may also be the biologically active form. Although the mutants Y53S and Y55K completely abrogate transport of the protein Pipe (which appears to be the primary function of Wind in the cell), only subtle differences can be seen in the putative Pipe-binding region. The Pipe binding in the active forms appears to involve hydrophobic interactions between aromatic systems, whereas the inactive mutants may be able to bind more strongly with the help of hydrogen bonds, which could disturb the delicate equilibrium required for effective Pipe transport. [ABSTRACT FROM AUTHOR]

The protein disulfide isomerase (PDI)-related protein Wind is essential in Drosophila melanogaster, and is required for correct targeting of Pipe, an essential Golgi transmembrane 2-O-sulfotransferase. Apart from a thioredoxin fold domain present in all PDI proteins, Wind also has a unique C-terminal D-domain found only in PDI-D proteins. Here, we show that Pipe processing requires dimeric Wind, which interacts directly with the soluble domain of Pipe in vitro, and we map an essential substrate binding site in Wind to the vicinity of an exposed cluster of tyrosines within the thioredoxin fold domain. In vitro, binding occurs to multiple sites within the Pipe polypeptide and shows specificity for two consecutive aromatic residues. A second site in Wind, formed by a cluster of residues within the D-domain, is likewise required for substrate processing. This domain, expressed separately, impairs Pipe processing by the full-length Wind protein, indicating competitive binding to substrate. Our data represent the most accurate map of a peptide binding site in a PDI-related protein available to date and directly show peptide specificity for a naturally occurring substrate. [ABSTRACT FROM AUTHOR]

In the developing Drosophila melanogaster embryo, dorsal-ventral patterning displays an absolute requirement for the product of the essential windbeutel gene, Wind. In homozygous windbeutel mutant flies, dorsalventral patterning fails to initiate because of the failure of the Golgi-resident proteoglycan-modifying protein, Pipe, to exit the endoplasmic reticulum, and this leads to the death of the embryo. Here, we describe the threedimensional structure of Wind at 1.9-Å resolution and identify a candidate surface for interaction with Pipe. This represents the first crystal structure of a eukaryotic protein-disulfide isomerase-related protein of the endoplasmic reticulum to be described. The dimeric protein is composed of an N-terminal thioredoxin domain and a C-terminal α-helical domain unique to protein-disulfide isomerase D proteins. Although Wind carries a CXXC motif that is partially surface accessible, this motif is redox inactive, and the cysteines are not required for the targeting of Pipe to the Golgi. However, both domains are required for targeting Pipe to the Golgi, and, although the mouse homologue ERp28 cannot replace the function of Wind, exchange of the Wind D-domain with that of ERp28 allows for efficient Golgi transport of Pipe. [ABSTRACT FROM AUTHOR]