[Two cases of granuloma annulare under anti-PD1 therapy]. / Deux observations de granulome annulaire sous anti-PD1.

BACKGROUND: Granuloma annulare as a granulomatous cutaneous reaction may be drug-induced. Immune checkpoint inhibitors including programmed death-1 (PD-1) inhibitors show remarkable antitumor activity and are approved for melanoma and other cancers. Different immune-related adverse effects have been described. We report herein a rare adverse effect of anti-PD1 therapy given for metastatic melanoma : granuloma annulare. PATIENTS AND METHODS: Two women receiving pembrolizumab metastatic melanoma presented with granuloma annulare. The therapy was continued in both cases. In the first patient, granuloma annulare appeared and then subsided; in the second patient, the lesion resolved completely with topical corticosteroids. DISCUSSION: While there have been reported cases of sarcoidosis induced by immunotherapies, immunotherapy-induced granuloma annulare has not been described. The pathogenesis of drug-induced granulomatous reactions is thought to involve autoimmune dysregulation affecting T cells, especially Th1 cells, which lead to granuloma formation. Granuloma annulare should thus be considered a cutaneous adverse effect of anti-PD1 immunotherapy.

[Iatrogenic skin lesions: a prospective study in premature infants born at less than 33 weeks of amenorrhea at Besançon university hospital]. / Lésions cutanées iatrogènes : étude prospective chez les prématurés de moins de 33 semaines de l'hôpital universitaire de Besançon.

BACKGROUND: Advances in neonatology have markedly improved prognosis for premature babies in recent years. However, they have also entailed the need for recourse to considerable intensive care involving potentially iatrogenic diagnostic and therapeutic acts. Among the resulting iatrogenic events, cutaneous lesions are the most frequent but have been the subject of very few studies. Our own study thus aimed to assess the rate of iatrogenic cutaneous events in premature infants born at less than 33 weeks of amenorrhea and hospitalised at Besançon university hospital and to identify the factors associated with the occurrence of these events. PATIENTS AND METHODS: This was a prospective study carried out in the department of paediatric intensive care and neonatology at Besançon university hospital between May 2011 and April 2012. All babies born before 33 weeks of amenorrhea hospitalised over this period were included. An iatrogenic event was defined as "an adverse event related to a medical procedure". Iatrogenic cutaneous events were reported to the dermatologist by medical and paramedical staff. RESULTS: One hundred and thirthteen newborn babies were included during the study period. Twenty-six iatrogenic cutaneous events were recorded in 19 infants, representing 16.8% of the population involved: nine were associated with ventilation techniques, six with the use of intravenous catheters, five with electrodes, two involved pressure sores, two were linked to the birth, one to disinfectants and one to dressings. The main risk factor was low birth weight (P=0.016). High prematurity and the duration of ventilation increased the risk, although not significantly. The death rate was higher in children with iatrogenic events but the difference was not significant. The duration of hospitalisation was unaffected by the presence or absence of an iatrogenic event. CONCLUSION: The frequency of iatrogenic cutaneous events is high in hospital departments in charge of very premature infants. Awareness by the medical and paramedical staff of the frequency of such iatrogenic events should improve the quality of care.

DLR MiroSurge: a versatile system for research in endoscopic telesurgery.

PURPOSE: Research on surgical robotics demands systems for evaluating scientific approaches. Such systems can be divided into dedicated and versatile systems. Dedicated systems are designed for a single surgical task or technique, whereas versatile systems are designed to be expandable and useful in multiple surgical applications. Versatile systems are often based on industrial robots, though, and because of this, are hardly suitable for close contact with humans. METHOD: To achieve a high degree of versatility the Miro robotic surgery platform (MRSP) consists of versatile components, dedicated front-ends towards surgery and configurable interfaces for the surgeon. RESULTS: This paper presents MiroSurge, a configuration of the MRSP that allows for bimanual endoscopic telesurgery with force feedback. CONCLUSIONS: While the components of the MiroSurge system are shown to fulfil the rigid design requirements for robotic telesurgery with force feedback, the system remains versatile, which is supposed to be a key issue for the further development and optimisation.

Multiple post-translational modifications of mouse insulin-like growth factor binding protein-6 expressed in epithelial Madin-Darby canine kidney cells.

Insulin-like growth factors (IGFs), IGF receptors and IGF binding proteins (IGFBPs) participate in the regulation of proliferation and differentiation of epithelial cells. Expression of the growth-inhibitory murine IGFBP-6 in epithelial Madin-Darby canine kidney (MDCK) cells followed by 2D analysis revealed the presence of multiple isoforms. Metabolic labelling experiments showed that several IGFBP-6 isoforms are modified by phosphate and sulfate groups. Expression analysis of mutant IGFBP-6 further demonstrated that serine residue 143 is O-glycosylated. Substitution of serine 143 by alanine did slightly reduce the preferential sorting of mIGFBP-6 to the apical site in MDCK cells grown on semipermeable filters. Both the presence of multiple and heterogeneously modified isoforms of murine IGFBP-6 in MDCK cells, and the preferential secretion of non-glycosylated IGFBP-6 mutants to the apical side suggest that the major apical sorting signal is the protein moiety.

Transferrin binds insulin-like growth factors and affects binding properties of insulin-like growth factor binding protein-3.

In the circulation, most of the insulin-like growth factors (IGFs) are bound to a ternary 150 kDa complex with IGF-binding protein (IGFBP)-3 and the acid labile subunit. In the current study, we identify transferrin (Tf) by mass spectrometry, and immunoprecipitation as a component of a major IGF-binding fraction separated from human plasma. IGF ligand blotting, cross-linkage experiments and surface plasmon resonance spectrometry have been used to demonstrate the capability of Tf to bind IGFs specifically. In combination with Tf, IGFBP-3 showed a 5-fold higher affinity for IGF-II than IGFBP-3 alone. The data suggest that Tf may play an important role in regulating IGF/IGFBP-3 functions.

MDCK cells secrete neutral proteases cleaving insulin-like growth factor-binding protein-2 to -6.

Proteolysis of insulin-like growth factor-binding proteins (IGFBPs) may be an important mechanism to regulate IGF availability and IGF-independent functions of IGFBPs. We analyzed the secretion of IGFBP proteases in Madin-Darby canine kidney (MDCK) cells. The results showed that several specific proteases were secreted, cleaving IGFBP-2 to -6 at neutral pH. The proteolytic activity against IGFBP-6 differed at least from IGFBP-5 protease activity in its sensitivity both to IGF-II and to the hydroxamic acid-based disintegrin metalloprotease inhibitor, as well as serine protease inhibitors. During partial purification steps, the serine protease inhibitor-sensitive fraction with IGFBP-6 protease activity was separated from fractions characterized by the presence of a 30-kDa disintegrin immunoreactive band. Whereas the IGFBP-4 and -6 proteases are predominantly secreted across the basolateral membrane, the majority of IGFBPs are sorted to the apical medium from filter-grown cells. These studies indicate that the side-specific secretion of several distinct IGFBP proteases with partially overlapping IGFBP specificities may be another level in the regulation of IGF-dependent epithelial functions.

Mouse insulin-like growth factor binding protein-6: expression, purification, characterization and histochemical localization.

The mitogenic and metabolic activities of insulin-like growth factors (IGF) are modulated by a family of six high affinity IGF binding proteins (IGFBPs). This study describes the expression of the mouse IGFBP-6 which is unique among IGFBPs in its preferential binding of IGF II, in insect cells using the baculovirus system. The purified, O-glycosylated IGFBP-6 was functional as shown by IGF binding and by inhibition of IGF II-stimulated DNA synthesis in human fibroblasts. Specific antibodies generated in chicken against the recombinant IGFBP-6 were used for Western blotting analysis and immunohistochemistry. Strong immunoreactivity was found in ossifying bones of the cranial base, in cell clusters of the pancreas anlage, in the trigeminal ganglion, on myoblasts, on motoneurons of the spinal cord of embryonic mice. In tissues of adult mouse, strong IGFBP-6 immunostaining was present in epidermal and peridermal layers of the skin, in meningeal layers, in long-striated skeletal muscle, and in the Langerhans' islets of the pancreas. No immunopositive staining was observed in lung and liver indicating that sites of synthesis and IGFBP action are different.

Proteolysis of insulin-like growth factor binding proteins by a novel 50-kilodalton metalloproteinase in human pregnancy serum.

Insulin-like growth factor binding proteins (IGFBP) proteases have been proposed to be involved in changes of serum IGFBP pattern during pregnancy. IGFBP-4 and -5 are degraded specifically by proteases in pregnancy serum in vitro, whereas IGFBP-3 proteolytic activity was also detected in nonpregnancy serum. To identify and characterize IGFBP proteases, human pregnancy serum was fractionated by size exclusion chromatography revealing IGFBP-4 protease activities in fractions coeluting with proteins of approximately 600-kDa and 50- to 100-kDa molecular mass. In both fractions, a predominant 50-kDa gelatinase was found, suggesting that parts of the gelatinase activity might aggregate or are complexed with other proteins forming a higher molecular complex. Hydroxyapatite chromatography and chromatofocusing of the 50- to 100-kDa serum fraction showed that the IGFBP-4 protease and the 50-kDa gelatinase activity were copurified. When the 50-kDa gelatinase-containing band was excised from the polyacrylamide gel, it exhibited IGFBP-4 proteolytic activity, resulting in the formation of 17- and 10-kDa fragments. [125I] IGFBP substrate zymography combined with fragment blotting showed that the 1,10-phenanthroline-sensitive 50-kDa protease activity purified by chromatofocusing also cleaved IGFBP-3 and -5. Other proteases detected in pregnancy serum fractions with Mr estimates of 79-, 30-, and 22-kDa degraded IGFBP-3 and -5 but not IGFBP-4. [125I] IGFBP-5 substrate zymography revealed that the 30-kDa IGFBP protease was inhibited by serine protease inhibitors. Whereas 1,10-phenanthroline inhibited the IGFBP proteolytic activity in the solution assay, serine protease inhibitors failed to affect proteolysis, indicating the predominant contribution of the metalloproteinase to IGFBP proteolysis. Tissue inhibitors of matrix metalloproteinases-1 and -2 revealed weak or no inhibition of IGFBP-4 and -5 proteolytic activity, whereas a hydroxamic acid-based inhibitor, potentially inhibiting disintegrin metalloproteases, completely prevented the proteolysis of IGFBPs. Whereas no specific immunoreactivity of the 50-kDa protein with antimatrix metalloproteinase-1, -2, -3, -9, or -13 antibodies was observed, antidisintegrin domain-specific antibodies bound to the 50-kDa gelatinase. These studies provide the first direct biochemical evidence that human pregnancy serum contains a 50-kDa IGFBP protease with properties of a soluble disintegrin metalloproteinase that appears to be potentially involved in regulating IGF bioavailability for placental and fetal growth.

Proteolysis of insulin-like growth factors (IGF) and IGF binding proteins by cathepsin D.

Various proteinases have been postulated to function in limited proteolysis of insulin-like growth factor binding proteins (IGFBPs) contributing to the regulation of IGF bioavailability. In this study, we report on the in vitro degradation of IGFs and IGFBPs by the purified acidic aspartylprotease cathepsin D that has been shown to proteolyze IGFBP-3. Recombinant human [125I] IGFBP-1 to -5 were processed by cathepsin D to fragments of defined sizes in a concentration dependent manner, whereas IGFBP-6 was not degraded. Ligand blotting revealed that none of the IGFBP-1 or -3 fragments formed by cathepsin D retain their ability to bind IGF. By N-terminal sequence analysis of nonglycosylated IGFBP-3 fragments produced by cathepsin D, at least four different cleavage sites were identified. Some of these cleavage sites were identical or differed by one amino acid from sites used by other IGFBP proteases described. The IGFBP-3 and -4 cleavage sites produced by cathepsin D are located in the nonconserved central region. IGF-I and -II, but not the unrelated platelet-derived growth factor BB, were degraded by cathepsin D in a time and concentration-dependent manner. We speculate that the major functional site of cathepsin D is intracellular and may be involved 1) in the selected clearance either of IGFBP or IGFs via different endocytic pathways or 2) in the general lysosomal inactivation of the IGF system.