Type I interferon response and vascular alteration in chilblain-like lesions during the COVID-19 outbreak.

BACKGROUND: The outbreak of chilblain-like lesions (CLL) during the COVID-19 pandemic has been reported extensively, potentially related to SARS-CoV-2 infection, yet its underlying pathophysiology is unclear. OBJECTIVES: To study skin and blood endothelial and immune system activation in CLL in comparison with healthy controls and seasonal chilblains (SC), defined as cold-induced sporadic chilblains occurring during 2015 and 2019 with exclusion of chilblain lupus. METHODS: This observational study was conducted during 9-16 April 2020 at Saint-Louis Hospital, Paris, France. All patients referred with CLL seen during this period of the COVID-19 pandemic were included in this study. We excluded patients with a history of chilblains or chilblain lupus. Fifty patients were included. RESULTS: Histological patterns were similar and transcriptomic signatures overlapped in both the CLL and SC groups, with type I interferon polarization and a cytotoxic-natural killer gene signature. CLL were characterized by higher IgA tissue deposition and more significant transcriptomic activation of complement and angiogenesis factors compared with SC. We observed in CLL a systemic immune response associated with IgA antineutrophil cytoplasmic antibodies in 73% of patients, and elevated type I interferon blood signature in comparison with healthy controls. Finally, using blood biomarkers related to endothelial dysfunction and activation, and to angiogenesis or endothelial progenitor cell mobilization, we confirmed endothelial dysfunction in CLL. CONCLUSIONS: Our findings support an activation loop in the skin in CLL associated with endothelial alteration and immune infiltration of cytotoxic and type I IFN-polarized cells leading to clinical manifestations.

Physicochemical characteristics of liposome encapsulation of stingless bees' propolis.

Nutraceuticals from natural sources have shown potential new leads in functional food products. Despite a broad range of health-promoting effects, these compounds are easily oxidized and unstable, making their utilization as nutraceutical ingredients limited. In this study, the encapsulated stingless bees' propolis in liposome was prepared using soy phosphatidylcholine and cholesterol by thin-film hydration technique. Three different formulations of phosphatidylcholine composition and cholesterol prepared by weight ratio was conducted to extract high propolis encapsulation. Physicochemical changes in the result of the encapsulation process are briefly discussed using scanning electron microscopy and Fourier Transform Infrared Spectroscopy. A dynamic light-scattering instrument was used to measure the hydrodynamic diameter, polydispersity index, and zeta potential. The increment of the liposomal size was observed when the concentration of extract loaded increased. In comparing three formulations, F2 (8:1 w/w) presented the best formulation as it yielded small nanoparticles of 275.9 nm with high encapsulation efficiency (66.9%). F1 (6:1 w/w) formed large particles of liposomes with 422.8 nm, while F3 (10:1 w/w) showed low encapsulation efficiency with (by) 38.7%. The liposome encapsulation will provide an effective nanocarrier system to protect and deliver the flavonoids extracted from stingless bees' propolis.

Lethal Morel-Lavallée lesion: A forensic radiology-pathology correlation.

Morel Lavallée lesion or closed degloving injury is normally associated with severe trauma and occurs when the skin and subcutaneous fatty tissue traumatically and abruptly separated from the underlying fascia thus creating a potential space filled with fluid. MVA is the commonest etiology but large or lethal Morel Lavallée is extremely rare. A 35 years old, female pillion rider was involved in a motor vehicle accident and sustained injuries to the left pelvis and thigh. Emergency laparotomy and intra-op abdominal and bilateral lower limb arteriogram revealed no significant finding. Her general condition and vital signs continued to deteriorate despite aggressive resuscitation and eventually died. Post-Mortem Computed Tomography and Post-Mortem Computed Tomography Angiogram was performed and revealed a large cavity in the left thigh suggestive of a lethal Morel Lavallée lesion. Findings were confirmed by conventional autopsy.

Sarcoidosis post-anti-PD-1 therapy, mimicking relapse of metastatic melanoma in a patient undergoing complete remission.

Novel treatments of metastatic melanoma include the usage of checkpoint inhibitors such as anti-cytotoxic-T-lymphocyte-associated antigen (CTLA-4) or anti-programmed death-1 (PD-1) antibodies, which are immunomodulatory antibodies that enhance the immune response against tumors. While they have substantially improved the prognosis for patients, these therapies are associated with a large spectrum of immune-related adverse effects (irAEs). We report a patient developing pulmonary and cutaneous sarcoidosis after pembrolizumab therapy while complete regression of stage IV melanoma was perceived. Coincident development of sarcoidosis and metastatic melanoma as illustrated by this case report poses a diagnostic challenge because, in contrast to other irAEs, both diseases share common features such as pulmonary lesions, hilar adenopathies and skin nodules. Considering sarcoidosis as a potential side-effect of pembrolizumab treatment is crucial to avoid misdiagnosis with malignant lesions, hence careful description and reporting of such cases is primordial for patient care and may also provide us with insightful knowledge on the pathogenesis of immune-related diseases.

RIPK1 both positively and negatively regulates RIPK3 oligomerization and necroptosis.

Necroptosis is a form of programmed cell death that depends on the activation of receptor interacting protein kinase-1 (RIPK1) and RIPK3 by receptors such as tumor necrosis factor (TNF) receptor-1. Structural studies indicate that activation of RIPK3 by RIPK1 involves the formation of oligomers via interactions of the RIP homotypic interaction motif (RHIM) domains shared by both proteins; however, the molecular mechanisms by which this occurs are not fully understood. To gain insight into this process, we constructed versions of RIPK3 that could be induced to dimerize or oligomerize in response to a synthetic drug. Using this system, we find that although the formation of RIPK3 dimers is itself insufficient to trigger cell death, this dimerization seeds a RHIM-dependent complex, the propagation and stability of which is controlled by caspase-8 and RIPK1. Consistent with this idea, we find that chemically enforced oligomerization of RIPK3 is sufficient to induce necroptosis, independent of the presence of the RHIM domain, TNF stimulation or RIPK1 activity. Further, although RIPK1 contributes to TNF-mediated RIPK3 activation, we find that RIPK1 intrinsically suppresses spontaneous RIPK3 activation in the cytosol by controlling RIPK3 oligomerization. Cells lacking RIPK1 undergo increased spontaneous RIPK3-dependent death on accumulation of the RIPK3 protein, while cells containing a chemically inhibited or catalytically inactive form of RIPK1 are protected from this form of death. Together, these data indicate that RIPK1 can activate RIPK3 in response to receptor signaling, but also acts as a negative regulator of spontaneous RIPK3 activation in the cytosol.

Trigeminocerebellar and trigemino-olivary projections in rats.

Retrograde and anterograde neuronal tracers (HRP, biocytin, biotinylated dextran-amine) were used to study the organisation of trigeminocerebellar and trigemino-olivary connections, focusing on the connectivity between trigeminal nuclear regions and the sagittal zones of the rat cerebellar cortex. Trigeminocerebellar projections were bilateral, but mostly ipsilateral. Direct trigeminocerebellar fibres originated mostly in the principal trigeminal nucleus (VP) and pars oralis (Vo), pars interpolaris (Vi), and to a lesser extent in pars caudalis (Vc) of the spinal trigeminal nucleus. Consistent projections were found from the Vc to cerebellar lobules IX and X. The trigeminal fibres terminated in the cerebellum in an organised fashion. The ventral part of the VP, Vo and Vi projected to the medial A zone and the C3 and D2 subzones, whereas the dorsal part of the nuclei projected to the lateral A zone and the C2, D0 and D1 subzones. In lobules IX and X, the organisation was different. The medial half of the VP, Vo, Vi and Vc projected to the lateral aspects of these lobules whereas their lateral part projected to their medial aspects. Trigeminal projections to the deep cerebellar nuclei were also present. Projections to a given sagittal zone concomitantly reached its corresponding nuclear target. Trigemino-olivary projections were principally contralateral. The Vo, Vi and Vc projected to the rostromedial dorsal accessory olive, the adjacent dorsal leaf and the dorsomedial part of the ventral leaf of the principal olive, which are known to project subzones C3, D0 and D1 of the rat cerebellar cortex.

Nucleus medialis-nucleus interpositus interface: its olivary and cerebellocortical projections in the rat.

The nuclear target of the X zone of the cerebellar cortex was identified in rats as clusters of neurons scattered at the interface between the nuclei medialis (NM) and interpositus (NI). In a previous study, we had outlined these target neurons and termed them "interstitial cell groups" (icg). In order to determine whether the icg should be considered as part of either the NM or the medial NI, we analyzed two efferent pathways from the icg: their nucleocortical and nucleoolivary projections. These were compared to their homologues from the NM and the NI. This analysis is based on mapping retrograde cell labeling and anterograde terminal labeling following microinjections of tracers in either the cerebellar cortex, the cerebellar nuclei, or the inferior olive. Nucleocortical projections originating from the icg are of the three types described previously: a "reciprocal" projection to the ipsilateral X zone, a "nonreciprocal" projection to the ipsilateral A zone, and a "symmetrical" projection to the contralateral X zone. These features can be considered as the summed characteristics of the nucleocortical projections from the NM and from the medial NI. Nucleoolivary projections from the icg target the lateral-rostral portion of the dorsal accessory olive as well as the centrocaudal part of the medial accessory olive. These pathways converge with the nucleoolivary projections from the medial NI and from the NM, respectively. The icg receives olivary afferents from both the regions of the dorsal and medial accessory olives to which it projects. On the basis of similarities shown here between the two types of efferents originating from the icg and those from the NM as well as the medial NI, the icg may be regarded as a "mosaic" of neuron clusters alternately belonging to the NM and the medial NI. Therefore, the icg would be reciprocally connected with the inferior olive.

Cerebellar afferences from the mesencephalic trigeminal nucleus in the rat.

The aim of the present study was to test for and characterize the organization of a direct projection from neurones of the trigeminal mesencephalic nucleus (Vme) to the cerebellum. WGA-HRP was used as a retrograde tracer following injections in the cerebellar cortex. The extent of each injection site within the sagittal zones was determined according to corticonuclear and olivocortical connections. Retrogradely labelled neurons were observed in the caudal part of the ipsilateral Vme only following vermal injection. The Vme projections reached exclusively the ipsilateral sagittal zone X in the anterior lobe, lobule VI and lobule IX. This identification was confirmed by anterograde labelling of mossy fibre terminals following a biocytin injection restricted to the Vme.

On the caudal extension of the X zone in the cerebellar cortex of the rat.

Following a selective injection of biotinylated dextran amine in the nuclear target (the interstitial cell groups, icg) of the X zone of the rat cerebellum, retrogradely labelled Purkinje cells (PCs) were found within a longitudinal strip of cortex, 250 microns in width, 1000 microns lateral to midline. This labelling delineates two compartments in the X zone, one rostral through lobules II-VI, and one caudal through lobules VIII-X. The whole rostrocaudal extent of the icg appears to be the target of PCs from both compartments without any apparent topographical organization.

The X zone and CX subzone of the cerebellum in the rat.

The existence of an X zone (lateral to the A zone) and a CX subzone (lateral to the C1 subzone) was documented within the anterior lobe and lobule VI in cats and primates. On the basis of their respective efferent and climbing fibre (CF) afferent connections, delineation of these two cortical subdivisions has been investigated here, in the rat, using small injections of WGA-HRP in the cerebellar cortex. We observe that both X and CX are "fractured" into a rostral and a caudal compartment. The rostral compartment of the X zone extends over lobules IV, V and VI and its caudal compartment over lobules VIII, IX and X. The rostral compartment of the CX subzone seems to be restricted to lobules V and VI, its caudal compartment cannot be topographically distinguished, over lobules IX and X, from the caudal compartment of the X zone. The olivary afferents to the X zone and the CX subzone arise from the horizontal and vertical lamellae of the medial accessory olive: subnucleus a projects into the rostral compartment and lobule VIII of the X zone. Subnuclei b and c project into the rostral compartment of both X and CX. The dorsomedial cell column selectively projects onto the caudal compartment of both X and CX over the vestibulo-cerebellum. The corticonuclear projections of the X zone have been found within the junctional region between the nucleus medialis and the nucleus interpositus (NI), here defined as the interstitial cell groups (icg), the corticonuclear projections of the CX subzone within the medial NI. It is suggested that the icg correspond to clusters of neurones dissociated from the medial aspect of the NI. We therefore consider the X zone and CX subzone of the rat, on the basis of their corticonuclear efferents, as "medial C1" and "lateral C1" subzone, respectively, although both may be regarded as part of the A zone on the basis of their olivary afferents.

Extraction of physostigmine from biologic fluids and analysis by liquid chromatography with electrochemical detection.

A rapid and simple method is described for the extraction of physostigmine (Phy) and its hydrolysis product, eseroline, from plasma, whole blood, and cerebrospinal fluid (CSF) and their subsequent quantitation by high-performance liquid chromatography (HPLC) with dual electrode electrochemical detection. Phy and eseroline were extracted from biologic fluids with cyano-phase columns eluted with 0.1 M citrate buffer, pH 4 containing 20% acetonitrile. Phy recovery from citrate buffer and CSF was nearly 100%. Phy recovery from plasma was 82% when methanol was used to precipitate proteins and 62% when HClO4 was used to precipitate proteins. Phy recovery from whole blood was only 17%. These results are discussed in the context of attempting to measure Phy in fluids of patients receiving this drug in clinical trials for the treatment of Alzheimer's disease.