Herpetiform pemphigus with characteristic transmission electron microscopic findings of various-sized ballooning vacuoles in keratinocytes without acantholysis.

We report a unique case of a Japanese woman with herpetiform pemphigus (HP) who had IgG autoantibodies reactive with nondesmosomal sites of keratinocytes and presented characteristic transmission electron microscopic (TEM) findings of various-sized vacuoles in keratinocytes without acantholysis. The patient presented with pruritic annular oedematous erythemas with small blisters lining the margins on the trunk and extremities. Histopathological examinations showed intraepidermal blisters with prominent infiltrations of eosinophils. Direct and indirect immunofluorescence tests revealed the presence of in vivo bound and circulating IgG autoantibodies to the keratinocyte cell surfaces. However, enzyme-linked immunosorbent assays for desmoglein (Dsg) 1, Dsg3 and desmocollins 1-3 showed negative results. Immunoblotting using the full-length human Dsg1 recombinant protein showed a positive band. TEM examination showed various-sized vacuoles squashing the nuclei in many keratinocytes, resulting in rupture of the cells. Immunoelectron microscopic examination revealed IgG deposition over the entire keratinocyte cell surfaces, which spared the desmosomes. IgG antibodies were also present on the inside walls of the vacuoles around the nuclei of keratinocytes and on the cell surfaces of infiltrating eosinophils. This patient also had marked eosinophilia and high levels of thymus and activation-regulated chemokine and interleukin-5 in the serum. These results indicated a novel autoantigen on the nondesmosomal keratinocyte cell surfaces and the pathogenesis of bullous spongiotic change with inflammation in HP.

Ir(iii) complex-based oxygen imaging of living cells and ocular fundus with a gated ICCD camera.

Phosphorescence lifetime imaging methods using oxygen-sensitive probes are very useful for visualizing the oxygen status of living cells and tissues with high spatial resolution. We aim to develop a useful oxygen detection technique combining a phosphorescent oxygen probe and an optimal detection method. Herein we present a biological oxygen imaging method using a microscope equipped with a gated intensified charge-coupled device (ICCD) camera as a detector and an Ir(iii) complex as a phosphorescent oxygen probe. Microscopic luminescence images of monolayer HT-29 cells (human colorectal adenocarcinoma cells) obtained using the cell-penetrating Ir(iii) complex BTPDM1 and an inverted microscope demonstrated that this method allowed visualization of the oxygen gradient produced in a monolayer of cultured cells when the monolayer is covered with a thin coverslip. Furthermore, combining the IR-emitting Ir(iii) complex DTTPH-PEG24 with a macrozoom microscope equipped with a gated ICCD camera enabled both the visualization of retinal vessels near the optic disc and the monitoring of oxygen level changes in a rabbit retina upon changing the inhaled oxygen content.

CNTN6 mutations are risk factors for abnormal auditory sensory perception in autism spectrum disorders.

Contactin genes CNTN5 and CNTN6 code for neuronal cell adhesion molecules that promote neurite outgrowth in sensory-motor neuronal pathways. Mutations of CNTN5 and CNTN6 have previously been reported in individuals with autism spectrum disorders (ASDs), but very little is known on their prevalence and clinical impact. In this study, we identified CNTN5 and CNTN6 deleterious variants in individuals with ASD. Among the carriers, a girl with ASD and attention-deficit/hyperactivity disorder was carrying five copies of CNTN5. For CNTN6, both deletions (6/1534 ASD vs 1/8936 controls; P=0.00006) and private coding sequence variants (18/501 ASD vs 535/33480 controls; P=0.0005) were enriched in individuals with ASD. Among the rare CNTN6 variants, two deletions were transmitted by fathers diagnosed with ASD, one stop mutation CNTN6W923X was transmitted by a mother to her two sons with ASD and one variant CNTN6P770L was found de novo in a boy with ASD. Clinical investigations of the patients carrying CNTN5 or CNTN6 variants showed that they were hypersensitive to sounds (a condition called hyperacusis) and displayed changes in wave latency within the auditory pathway. These results reinforce the hypothesis of abnormal neuronal connectivity in the pathophysiology of ASD and shed new light on the genes that increase risk for abnormal sensory perception in ASD.

Lichen amyloidosus as a sweat gland/duct-related disorder: resolution associated with restoration of sweating disturbance.

BACKGROUND: Although a number of pathological processes resulting in amyloid deposition have been described in lichen amyloidosus (LA), no attention has been paid to the involvement of sweat glands/ducts in the pathogenesis of LA. According to recent studies, follicular structures are usually spared in serial histological sections of LA, and deposits of amyloid are likely to be confined to areas that display xerosis, suggesting that decreases in skin wetness by sweating disturbance seem to initiate LA. OBJECTIVES: To investigate whether sweating disturbance could represent an early event that triggers LA, and whether resolution of LA could be induced by restoring the sweating disturbance. METHODS: By using the impression mould technique, which allows an accurate quantification of individual sweat glands/ducts actively delivering sweat, we examined sweat responses to thermal stimulus in LA lesions before and after treatment with a moisturizer. RESULTS: Sweating disturbance was most profoundly detected in the 'hub' structure of the LA papule, and this disturbance due to leakage of sweat could be restored by short-term treatment with a moisturizer, particularly when used under occlusion. CONCLUSIONS: This study was limited by the relatively small sample size. Treatment of LA should be primarily directed at preventing leakage of sweat into the dermis or epidermis and therefore sweat delivery to the skin surface could be made easier.

Contactin-5 expression during development and wiring of the thalamocortical system.

The gene encoding the neural cell adhesion molecule Cntn5 (a.k.a. NB-2) has been put forward as a candidate in neurodevelopmental disorders, like autism spectrum disorder (ASD), by recent genetic findings. Little is known about the expression pattern and function of the gene, and its functional involvement in brain development has remained elusive. So far, most research has focused on its early postnatal expression in the auditory system, where the absence of Cntn5 causes abnormal responses to acoustic stimuli and a decrease in fiber density. The current study shows that the Cntn5 gene is expressed in forebrain structures during embryonic development, starting at E15.5, and that it continues to be expressed into adulthood. Sites of strong expression included the thalamus, the caudate putamen (CPu) and to a lesser extent layer Va of the cerebral cortex. Cntn5-positive thalamic nuclei include the laterodorsal (LD), ventrolateral (VL) and posterior group (Po), which contain glutamatergic neurons. Visualization of the expression pattern through the Tau-LacZ fusion protein coded by an insert in the Cntn5 gene, demonstrated that Cntn5-positive nuclei of the thalamus project to the cortex, based on co-localization with thalamocortical markers L1 and Calretinin. These results indicate that the cell adhesion functions of Cntn5 are exploited for circuit formation and connectivity in early development and for synaptic maintenance during adulthood. Subtle alterations in the formation of the thalamocortical circuit may contribute to neurodevelopmental disorders, such as ASD.

Effects of hydrodynamic interaction on the equivalent conductivity minimum of electrolyte solutions in solvents of low dielectric constant.

Brownian dynamics simulation on model electrolyte solutions in our previous work [T. Yamaguchi et al., J. Chem. Phys. 134, 244506 (2011)] is extended to include the hydrodynamic interaction between ions, in order to examine its effects on ionic mobility in solvents of low dielectric constant. The effects of the hydrodynamic interaction are rather small as a whole, and the equivalent conductivity minimum is observed in systems with the hydrodynamic interaction. The hydrodynamic interaction increases the self-diffusion coefficient while decreases the equivalent conductivity, thereby increases the deviation from the Nernst-Einstein relationship. Based on the analysis of the time-dependent ionic mobilities, these changes are elucidated in terms of the electrophoretic and relaxation effects. It is also demonstrated that the concentration dependence of the ionic mobilities with the hydrodynamic interaction is reproduced fairly well by a theoretical calculation.

Phase diagram of interacting composite fermions in the bilayer nu=2/3 quantum hall effect.

We study the phase diagram of composite fermions (CFs) in the presence of spin and pseudospin degrees of freedom in the bilayer nu=2/3 quantum Hall (QH) state. Activation studies elucidate the existence of three different QH states with two different types of hysteresis in the magnetotransport. While a noninteracting CF model provides a qualitative account of the phase diagram, the observed renormalization of tunneling gap and a non-QH state at high densities are not explained in the noninteracting CF model, and are suggested to be manifestations of interactions between CFs.

Antigens of monoclonal antibody NB3C4 are novel markers for oligodendrocytes.

We produced NB3C4, a novel monoclonal antibody specific for oligodendrocytes, using human neuroblastoma IMR-32 cells. NB3C4 specifically recognized oligodendrocytes in the CNS, although it bound to neuroblastoma IMR-32 cells and oligodendrocytes in vitro. Double immunofluorescence staining of rat brain using NB3C4 and anti-GST-pi, anti-glial fibrillary acidic protein (GFAP), or anti-neurofilament 200 (NF) antibody revealed that anti-GST-pi antibody identified an oligodendrocyte marker recognizing NB3C4-positive cells, while both anti-GFAP and anti-NF antibody did not. Western blotting of rat brain homogenates showed that NB3C4 bound three proteins of 22-28 kDa, while the anti-GST-pi recognized a 27 kDa protein. Therefore, antigens recognized by NB3C4 could be novel markers for oligodendrocytes.

Expression of glycoconjugates bearing the Lewis X epitope during neural differentiation of P19 EC cells.

The Lewis X (Le(x)) bearing glycolipids were noticeably increased in amounts during the course of neural differentiation of P19 EC cells induced by retinoic acid (RA, all-trans form). Applying neoglycolipid technology and in situ TLC-LSIMS, the oligosaccharide chains of these scarce Le(x) bearing glycolipids were partially characterized after released by endoglycoceramidase and subsequent conversion into neoglycolipids. In order to understand the enzymatic basis for the expression of Le(x) bearing glycolipids, we measured glycolipid, glycoprotein and oligosaccharide fucosyltransferase (Fuc-T) activities using appropriate substrates in P19 EC cells with or without RA treatment. All three Fuc-Ts were increased after RA treatment and the highest activity was in the differentiated neural cells. We then investigated the two possible Fuc-T genes that might be responsible for these changes using RT-PCR analysis. Mouse Fuc-TIX (mFuc-TIX) transcript was detected in all cell types but it was only strongly expressed in RA-induced aggregates and neural cells. In the case of mouse Fuc-TIV (mFuc-TIV) gene, its transcript was only detectable in RA-induced aggregates and not found in either undifferentiated or RA-induced neural cells. These results strongly support that RA induces only a transient expression of the mFuc-TIV gene in cell aggregates but a more persistent expression of the mFuc-TIX gene at the transcription level throughout neural cell differentiation. The mFuc-TIX gene is probably the main cause for the increased expression of Le(x) glycoconjugates during neural differentiation of P19 EC cells.