Oral administration of milk-derived phospholipids inhibits penetration of cutaneous nerve fibres into epidermis in a mouse model of acute dry skin.

The density of intraepidermal nerve fibres has been shown to be higher in itchy dry skin than in healthy skin, suggesting that epidermal hyperinnervation is at least partly involved in peripheral itch sensitization. We investigated whether oral administration of milk-derived phospholipids (MPLs) would inhibit epidermal hyperinnervation in a mouse model of dry skin. We found that the number of intraepidermal nerve fibres was significantly lower in the MPL group than in the control group. Expression of nerve growth factor (NGF) levels in the epidermis was significantly decreased by oral administration of MPLs, whereas expression of semaphorin (Sema)3A, a nerve repulsion factor, was increased in the MPL group. These results suggest that dietary MPLs attenuate the penetration of nerve fibres into the epidermis by reducing epidermal NGF levels and increasing Sema3A level. Thus, dietary MPLs may have beneficial effects in the prevention and/or alleviation of dry skin-induced itch by reducing intraepidermal nerve fibre density.

Antibody against chromatin assembly factor-1 is a novel autoantibody specifically recognized in systemic lupus erythematosus.

Autoantibodies to proliferating cell nuclear antigen (PCNA) are specifically, if rarely, present in systemic lupus erythematosus (SLE) patient sera. Even SLE patients lacking PCNA reactivity often show reaction to PCNA-binding protein. Here, immunoreactivity to chromatin assembly factor-1 (CAF-1), an essential molecule for DNA replication and a PCNA-binding protein, was compared for the sera of SLE patients, normal healthy controls (NHCs) and other disease controls, and in autoimmune sera reactive to standard autoantigens, by enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescence, and immunoblotting. CAF1 and IRF1 expression in SLE and NHC peripheral mononuclear cells were compared by quantitative real-time polymerase chain reaction. Serum interferon-γ-inducing protein-10 and anti-double-stranded (ds)DNA antibody levels were measured by ELISA. Increased CAF-1 autoimmune reactivity was recognized in SLE or serum anti-dsDNA antibody-positive patients. Significantly greater central nervous system (CNS) involvement (aseptic meningitis) and serum anti-dsDNA antibody titers were present more often in anti-CAF-1 antibody-positive than antibody-negative SLE patients. IFN-γ positively regulated CAF-1 expression in vitro and was associated with anti-CAF-1 antibody production in SLE. Thus, a novel anti-CAF-1 autoantibody is frequently found in patients with SLE and is a useful biomarker for diagnosis, especially in cases with CNS involvement. Aberrant IFN-γ regulation appears to play an important role in anti-CAF-1 antibody production in SLE.

Neurotropin inhibits the increase in intraepidermal nerve density in the acetone-treated dry-skin mouse model.

Epidermal hyperinnervation is considered one cause of sensitization to itch, and is thought to regulated by keratinocyte-derived axonal guidance molecules, including nerve growth factor (NGF) and semaphorin (Sema)3A. Neurotropin (NTP) shows antipruritic effects in allergic disease and is also used for pain relief. Using cultured rat dorsal root ganglion neurones, we previously found that NTP inhibited NGF-induced neurite outgrowth. However, no such inhibitory effect has been shown in vivo. We therefore assessed the effects of intraperitoneal administration of NTP on nerve density and expression of NGF and Sema3A mRNAs in the epidermis of acetone-treated mice showing epidermal hyperinnervation. We found that NTP significantly reduced intraepidermal nerve growth in these acetone-treated mice. NTP significantly upregulated epidermal Sema3A mRNA, but had no effect on expression of epidermal NGF mRNA. These findings indicate that NTP may reduce intraepidermal nerve density by inducing expression of Sema3A in the epidermis.

Possible role of the JAK/STAT pathways in the regulation of T cell-interferon related genes in systemic lupus erythematosus.

Changes in gene expression in CD3+ T cells associated with disease progression in systemic lupus erythematosus (SLE) patients were determined. The genes related to SLE disease-related activities were identified and their gene regulatory networks were investigated. Analyses of gene expression were performed by both DNA microarray and real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). The expression of certain genes including interferon (IFN) regulatory factor (IRF)-related genes, such as IFN-regulated, -related, and -signature genes was increased in the active phase of SLE. Pathway network analyses suggested that these IRF-related genes are regulated through the Janus kinase/signal transducer and activator of transcription (JAK/STAT) pathway. JAK/STAT pathway-mediated regulation of IRF-related genes may have an important role in the disease activity of SLE. Inhibitors of JAK/STAT cascade may be useful as therapeutic agents.

Neurotropin inhibits both capsaicin-induced substance P release and nerve growth factor-induced neurite outgrowth in cultured rat dorsal root ganglion neurones.

BACKGROUND: Neurotropin (NTP), a biological extract from rabbit skin inoculated with vaccinia virus, is an effective analgesic and anti-allergic agent, and has antipruritic effects in various dermatoses including eczema, dermatitis and urticaria. In patients receiving haemodialysis who have pruritus, NTP appears to exert its antipruritic effect by lowering the plasma levels of substance P (SP), but its underlying mechanisms are not fully understood. AIM: To investigate the antipruritic mechanisms of NTP. METHODS: The effects of NTP on capsaicin-induced SP release from neonatal rat dorsal root ganglion (DRG) neurones were assessed by measuring SP concentrations in culture media by a competitive ELISA. The effects of NTP on nerve growth factor (NGF)-induced neurite outgrowth were assessed by measuring the length of the longest process of cultured DRG neurones. The neuronal cytotoxicity of NTP was determined using a methylthiazole tetrazolium cytotoxicity assay. RESULTS: NTP dose-dependently inhibited capsaicin-induced release of SP from cultured DRG neurones, whereas NTP alone had no effect on SP release. Moreover, NTP dose-dependently inhibited NGF-induced neurite outgrowth in cultured DRG neurones. NTP had no observable cytotoxicity. CONCLUSIONS: These results suggest that NTP exerts its antipruritic effects by inhibiting both SP release and neurite outgrowth of cutaneous sensory nerves.

In vitro model for penetration of sensory nerve fibres on a Matrigel basement membrane: implications for possible application to intractable pruritus.

BACKGROUND: Epidermal hyperinnervation occurs in dermatoses with intractable pruritus, such as atopic dermatitis, suggesting that the hyperinnervation is partly responsible for abnormal itch perception. OBJECTIVES: To investigate the mechanisms of penetration of sensory nerve fibres into the basement membrane of the skin. METHODS: A rat dorsal root ganglion neurone culture system consisting of Matrigel and a Boyden chamber containing a nerve growth factor (NGF) concentration gradient was used. In some experiments, matrix metalloproteinase (MMP) blockers and semaphorin 3A (Sema3A) were added to the culture system. Matrigel-coated membranes were stained with anti-Tau antibody, and the number of nerve fibres that crossed the membrane was counted. Expression of MMPs in the cultured neurones was examined at mRNA and protein levels by quantitative reverse transcription-polymerase chain reaction and immunocytochemistry, respectively. The activity was also examined by zymography. RESULTS: Nerve fibres penetrated into Matrigel in the presence of an NGF concentration gradient, which was dose-dependently inhibited by GM6001, a broad-spectrum MMP inhibitor. Transcripts for MMP2, but not MMP9, were increased in the cultured neurones, and the penetration was dose-dependently inhibited by MMP-2 blockers. MMP-2 and its activity were partially localized on the NGF-responsive growth cones. NGF also upregulated pro-MMP-2 activation molecules in the cultured neurones. Sema3A stimulation showed the opposite effects on these NGF-dependent events. Interestingly, MMP2 expression was modulated by extracellular matrix (ECM) substrates for this enzyme. CONCLUSIONS: Membrane-associated MMP-2 contributes to penetration of nerve fibres into Matrigel through modulation by axonal guidance molecules and/or ECM. These findings provide insight for understanding the development of intractable pruritus involving epidermal nerve density.

Changes in the gene expression of peripheral blood mononuclear cells during the menstrual cycle of females is associated with a gender bias in the incidence of systemic lupus erythematosus.

OBJECTIVE: The incidence of systemic lupus erythematosus (SLE) is far higher in females than in males and the onset and/or disease activity is influenced by pregnancy and the menstrual cycle. Sex hormones seem to influence the pathogenesis of SLE, therefore, changes in gene expression in peripheral blood mononuclear cells (PBMC) were examined during the menstrual cycle in females, under the comparison of gene expression of patients with SLE. METHODS: The detection and a quantitative analysis of the gene expression was performed by DNA microarray or real-time quantitative polymerase chain reaction (RQ-PCR) method. RESULTS: There were thirteen known genes which showed significant quantitative changes during the menstrual cycles of females, but not in males. Among these genes, statistical quantitative differences between normal controls and SLE patients were observed in six genes. CONCLUSION: Based on these findings, certain genes (such as the tumor necrosis factor receptor superfamily, member 14; TNFRSF14, and signal regulatory protein, gamma; SIRPG) appear to contribute to gender difference of SLE.

Protein biomarker analysis by mass spectrometry in patients with rheumatoid arthritis receiving anti-tumor necrosis factor-alpha antibody therapy.

OBJECTIVE: To investigate the mechanism of action of anti-tumor necrosis factor-alpha (TNF-alpha) antibody in patients with rheumatoid arthritis (RA), we analyzed serum or plasma proteins by mass spectrometry system. METHODS: Ten RA patients who received treatment with anti-TNF-alpha antibody were studied. Samples obtained before and after therapy were analyzed by a two-dimensional liquid chromatography tandem mass spectrometry (2D LC-MS/MS) system after pretreatment by a recently developed method to remove high molecular weight proteins. RESULTS: Using this system, certain proteins were identified after treatment with anti-TNF-alpha antibody, including proteins related to the TNF-alpha-mediated pathway for nuclear factor kappa B (NF-kappaB) activation and/or to the metabolism (including regeneration) of articular cartilage. CONCLUSION: Our mass spectrometry system appears to be useful for proteomic analysis. The efficacy of anti-TNF-alpha antibody therapy for RA may be related to various consequence of the inhibition of TNF-alpha activity.

Epithelial fibroblast growth factor receptor 1 regulates enamel formation.

The interaction between epithelial and mesenchymal tissues plays a critical role in the development of organs such as teeth, lungs, and hair. During tooth development, fibroblast growth factor (FGF) signaling is critical for regulating reciprocal epithelial and mesenchymal interactions. FGF signaling requires FGF ligands and their receptors (FGFRs). In this study, we investigated the role of epithelial FGF signaling in tooth development, using the Cre-loxp system to create tissue-specific inactivation of Fgfr1 in mice. In K14-Cre;Fgfr1(fl/fl) mice, the apical sides of enamel-secreting ameloblasts failed to adhere properly to each other, although ameloblast differentiation was unaffected at early stages. Prior to eruption, enamel structure was compromised in the K14-Cre;Fgfr1(fl/fl) mice and displayed severe enamel defects that mimic amelogenesis imperfecta (AI), with a rough, irregular enamel surface. These results suggest that there is a cell-autonomous requirement for FGF signaling in the dental epithelium during enamel formation. Loss of Fgfr1 affects ameloblast organization at the enamel-secretory stage and, hence, the formation of enamel.

Aldo-keto reductase family 1, member B10 in uterine carcinomas: a potential risk factor of recurrence after surgical therapy in cervical cancer.

Aldo-keto reductase family 1, member B10 (AKR1B10), an enzyme that converts retinals into retinols is known to detect in non-small cell lung carcinoma (squamous cell- and adeno-carcinomas), but is barely expressed in normal tissues. Since these types of carcinoma occur frequently in the uterus (like in the lung), AKR1B10 may also be overexpressed in two major types of uterine cancer, cervical cancer (CC), and endometrial cancer (EMC). The objective of this study is to investigate AKR1B10 expression in uterine cancer and to analyze its clinical significance. In samples from uterine cancer patients, AKR1B10 was detected in 6 out of 30 (20.0%) CC cases and 6 out of 38 (15.8%) EMC cases. Statistical analysis indicated that AKR1B10 expression was associated with tumor recurrence after surgery and keratinization of squamous cell carcinoma only in CC. Although retinol (a metabolic product by AKR1B10) was observed in the normal epithelium, the molecule was not observed in cancer cells of AKR1B10-positive CC samples suggesting that the recurrence in CC may not depend on the convert of retinals into retinols via AKR1B10, a potential indicator in the management of patients with CC.

A case of subcutaneous phaeohyphomycotic cyst due to Exophiala jeanselmei complicated with systemic lupus erythematosus.

We report a case of subcutaneous phaeohyphomycosis by Exophiala jeanselmei that appeared on the extensor surface of the left lower leg of a 34-year-old woman with systemic lupus erythematosus (SLE). The superficial symptoms were a subcutaneous nodule 2.5 x 2 cm in size discharging a serous exudate from its center. Histopathological examination revealed granulomatous changes including large numbers of neutrophils in the dermis and the subcutaneous tissues. In addition, periodic acid-Schiff-positive fungal elements consisting of many yeast-like cells and chains of cells with hyphae were seen. The statistics on E. jeanselmei infections in Japan indicated that 54 cases (24 in men and 30 in women) had been reported, of which 50 (21 in men and 29 in women) were phaeohyphomycosis, and about half had underlying diseases; and the sites of the lesions were mainly on the extremities.

Effect of ACTH on the imipramine- and desipramine-induced decrease in duration of immobility time as measured in a rat forced swimming test.

In a rat forced swimming test (FS), we examined the effect of repeated injections of ACTH (adrenocorticotropic hormone) for 14 days on the decreased duration of immobility time produced by imipramine and desipramine. Both imipramine (15 and 30 mg/kg, p.o.) and desipramine (15 and 30 mg/kg, p.o.) significantly decreased the duration of immobility time in the FS. On the other hand, ACTH (100 microg/kg, i.p.) alone did not affect the duration of immobility time in FS. When ACTH (100 microg/kg, i.p.) was injected for 14 days before the 15-min swim session, it counteracted the decreased duration of immobility time induced by both imipramine and desipramine. Thus, ACTH seems to play a key role in decreasing the duration of immobility time of antidepressants in this test.

Repeated treatment with imipramine, fluvoxamine and tranylcypromine decreases the number of escape failures by activating dopaminergic systems in a rat learned helplessness test.

Chronic administration of antidepressants has been shown to reduce the number of escape failures in the rat learned helplessness test (LH). In the present study we investigated the role of D1, D2 and D3 receptors in mediating this effect. In our first series of experiments, we demonstrated that SKF38393, D1 receptor agonist, in a dose of 2.5 mg/kg (i.p.) and quinpirole, D2 receptor agonist in a dose of 0.5 mg/kg (i.p.), significantly decreased the number of escape failures in LH, and these were reversed by SCH23390 (0.015 mg/kg), D1 receptor antagonist, and by sulpiride (25 mg/kg), D2 receptor antagonist, respectively. In contrast, 7-OH-DPAT, a D3 receptor agonist, in a dose of 10 mg/kg (i.p.) did not affect the number of escape failures in LH. In a second series of experiments, we showed that eight days of repeated treatment with imipramine (10 mg/kg, p.o.), fluvoxamine (1.25 mg/kg, p.o.) and tranylcypromine (1.25 mg/kg, p.o.) significantly decreased the number of escape failures in LH. The decrease in escape failures seen with use of imipramine and tranylcypromine was reversed by sulpiride in LH, but not by SCH23390. On the other hand, the effect of fluvoxamine was reversed by both SCH23390 and sulpiride. These findings indicate that stimulation of D1 and D2 receptors decreased the number of escape failures in LH, respectively. Thus, D2 and/or D1 receptors are probably involved in the decreased number of escape failures in case of repeated treatment with antidepressants in LH.

Detection of occlusal caries under sealants by use of a laser fluorescence system.

OBJECTIVE: In recent years, various dental caries diagnosis systems, including digital radiography, light fluorescence, and lasers, have been developed. The aim of this study was to evaluate the new laser fluorescence system known as DIAGNOdent for its ability to detect occlusal caries under sealants. BACKGROUND DATA: Fluorescence induced by laser light for early diagnosis of enamel caries was introduced by Bjelkhagen et al. in 1982, but these systems were difficult to use in the oral cavity. Recently, DIAGNOdent has been shown to have a higher diagnostic validity for the detection and quantification of caries lesions than the electronic caries monitor and to have a higher reproducibility for measurements, according to work by Lussi et al. in 1999. The laser diagnosis system is able to indicate the value of reflected fluorescence on a digital display. This value is associated with the carious progression. METHODS: Thirty-two extracted permanent premolars and molars that had occlusal dental caries were used in this study. The reflected fluorescence was measured by use of DIAGNOdent at the occlusal surface of the tooth before and after chemical irrigation, acid etching, and application of a light-cured fluoride releasing sealant. The measurements were two or three points per experimental tooth. A total of 53 points on the experimental teeth was evaluated. RESULTS: The mean reflected fluorescence value was significantly decreased by chemical irrigation; however, no difference was noted between the value before and the salve after acid etching. These teeth were divided into three groups: clear, red, and white as tooth color sealants of the tooth. The reflected fluorescence value before and after sealants was tested for statistical significance. The diagnosis of caries under sealants was up to 10 of reflected fluorescence value in the study. Clear and red sealants were used to reveal caries under sealants using visual inspection, and all sealants groups achieved 90% of reflected fluorescence value after sealants application. However, the white sealants did not reveal caries through visual inspection. The detection ability of caries under white sealants was 53.5% (n = 23/42). CONCLUSIONS: This laser diagnosis system makes it easy to detect the existence of caries under a pit and fissure sealant during a routine check-up.

[A case of mandibular gingival cancer T4N0M0 which markedly responded to a combined therapy of nedaplatin with 5-fluorouracil].

We recently experienced a case of mandibular gingival cancer T4N0M0 which markedly responded to a combination therapy of nedaplatin (254-S) with 5-fluorouracil (5-FU). The patient was a 68-year-old male who visited our department with the main complaint of ulceration in the left mandibular gingiva. Biopsy revealed a moderately differentiated squamous cell carcinoma which extended to the mandible, mandibular gingiva, buccal mucosa, half tongue and oral floor on the left side of the face. As a neoadjuvant chemotherapy (NAC), 254-S at a dose of 100 mg/m2 was intravenously administered on day 1, while 5-FU at a dose of 700 mg/m2/day was intravenously administered from day 1 to 5 in succession. Hydration (2,000 ml/day) was performed from day 1 to 3. Adverse reactions observed included thrombocytopenia, anorexia, nausea, vomiting, stomatitis and SIADH, but no sign of renal dysfunction was observed. The clinical outcome was evaluated as CR. Surgery was performed later. Pathological examination of the extracted tissues showed tumor cells in the tongue only, indicating an excellent effect of this combination therapy of 254-S and 5-FU.

Availability of learned helplessness test as a model of depression compared to a forced swimming test in rats.

This study was designed to evaluate the antidepressant activity of various antidepressants using the learned helplessness test (LH) or the forced swimming test (FS) in rats. Repeated treatment of the tricyclic antidepressants imipramine (10 mg/kg, p.o.), clomipramine (0.625 mg/kg, p.o.), amitriptyline (10 mg/kg, p.o.) and amoxapine (20 mg/kg, p.o.) reduced the number of escape failures in the LH group, respectively. Repeated treatment of an atypical antidepressant, mianserin (2.5 and 5 mg/kg, p.o.), and one of the selective serotonin reuptake inhibitors (SSRI), fluvoxamine (1.25 mg/kg, p.o.), also reduced the number of escape failures in the LH group. In the FS, repeated treatment of imipramine (5, 10 mg/kg, p.o.), amitriptyline (5, 10 mg/kg, p.o.) and mianserin (10 mg/kg) significantly decreased the duration of immobility time. On the other hand, repeated treatment of amoxapine (5-20 mg/kg), clomipramine (0.1325-1.25 mg/kg, p.o.) and fluvoxamine (0.3125-1.25 mg/kg, p.o.) failed to decrease the duration of immobility time in the FS group. In conclusion, these results suggest that the LH group is sensitive to agents with a variety of antidepressant properties compared to the FS group in rats.