Development of immune and microbial environments is independently regulated in the mammary gland.

Breastfeeding is important for mammals, providing immunological and microbiological advantages to neonates, together with the nutritional supply from the mother. However, the mechanisms of this functional diversity in the mammary gland remain poorly characterized. Here, we show that, similar to the gastrointestinal tract, the mammary gland develops immune and microbial environments consisting of immunoglobulin A (IgA) and the microflora, respectively, both of which are important for protecting neonates and the mother from infectious diseases. The IgA production and microflora development are coordinated in the gastrointestinal tract but seem to be independently regulated in the mammary gland. In particular, the chemokine (C-C motif) ligand 28 and poly-Ig receptor, crucial molecules for the IgA production in milk, were expressed normally in germ-free lactating mice but were almost undetectable in postweaning mothers, regardless of the microflora presence. Our findings offer insights into potentially improving the quality of breastfeeding, using both immunological and microbiological approaches.

Interleukin 33 Is Induced by Tumor Necrosis Factor α and Interferon ƴ in Keratinocytes and Contributes to Allergic Contact Dermatitis

Background: Interleukin (IL) 33, a novel member of the IL-1 family, is produced mainly by epithelial cells and endothelial cells in response to various types of stress, including necrosis. The effects of IL-33 on the immune cells involved in allergic contact dermatitis have recently been revealed in vitro. However, in vivo, the induction mechanism and function of IL-33 are not fully understood. Objectives: Our objectives were to investigate induction of IL-33 in keratinocytes and to evaluate the functions of IL-33 and its inducers in a murine model of allergic contact dermatitis. Material and Methods: KERTr cells, a human keratinocyte cell line, were cultured with various cytokines, including tumor necrosis factor (TNF) α and interferon (IFN) ƴ. IL-33 expression was detected using quantitative reverse transcriptase polymerase chain reaction, immunocytochemistry, and Western blotting. The functions of IL-33, TNF- α, and IFN-􀁡 in allergic contact dermatitis were evaluated using a murine model. Results: TNF- α and IFN- ƴ induced expression of IL-33 mRNA and protein in KERTr cells. Blockade of IL-33 attenuated swelling in the ears of the experimental mice. Similar effects were noted for blockade of TNF- α and IFN- ƴ in these mice. Conclusions: TNF- α and IFN- ƴ induce expression of IL-33, and IL-33 produced by keratinocytes contributes to allergic contact dermatitis. Blockade of IL-33, TNF- α, and IFN- ƴ could represent novel and potent strategies to treat allergic contact dermatitis (AU)

Antecedentes: La Interleucina 33 (IL-33), un nuevo miembro de la familia de la IL-1, es producida fundamentalmente por las células epiteliales y endoteliales en respuesta a diferentes estímulos, incluyendo la necrosis. Recientemente se han confirmado los efectos de esta IL sobre las células del sistema inmunológico in vitro en pacientes con dermatitis de contacto, aunque los mecanismos y función in vivo de la IL-33 no son bien conocidos. Objetivos: El objetivo de este estudio fue analizar los factores que podrían inducir IL-33 en queratinocitos y evaluar las funciones de esta citocina y de sus inductores en un modelo murino de dermatitis alérgica de contacto. Métodos: Para ello se cultivaron células KERTr, una línea celular de queratinocitos humanos, en presencia de varias citocinas, incluyendo TNF- α e IFN − ƴ. La expresión de IL-13 se detectó mediante PCR cuantitativa a tiempo real, inmunocitoquímica e inmunobloting. Así mismo se evaluó la función de IL-33, TNF- α, e IFN- ƴ en el modelo murino. Resultados: En cuanto a los resultados obtenidos TNF- α y IFN- ƴ indujeron la expresión de mRNA y expresión de proteína en las células KERTr. El bloqueo de IL-33 atenúa la inflación en la dermatitis de contacto murina. Efectos similares se obtienen mediante el bloqueo de TNF- α y IFN- ƴ. Conclusiones: En conclusión, TNF- α and IFN- ƴ son inductores de la producción de IL-33, y además esta citocina producida por los queratinocitos contribuye a la expresión de dermatitis alérgica de contacto. El bloqueo de no solo IL-33, sino también de TNF- α y IFN- ƴ podría representar una modalidad terapéutica nueva y potente en la dermatitis alérgica de contacto (AU)

Interleukin 33 is induced by tumor necrosis factor alpha and interferon gamma in keratinocytes and contributes to allergic contact dermatitis.

BACKGROUND: Interleukin (IL) 33, a novel member of the IL-1 family, is produced mainly by epithelial cells and endothelial cells in response to various types of stress, including necrosis. The effects of IL-33 on the immune cells involved in allergic contact dermatitis have recently been revealed in vitro. However, in vivo, the induction mechanism and function of IL-33 are not fully understood. OBJECTIVES: Our objectives were to investigate induction of IL-33 in keratinocytes and to evaluate the functions of IL-33 and its inducers in a murine model of allergic contact dermatitis. MATERIAL AND METHODS: KERTr cells, a human keratinocyte cell line, were cultured with various cytokines, including tumor necrosis factor (TNF) alpha and interferon (IFN) gamma. IL-33 expression was detected using quantitative reverse transcriptase polymerase chain reaction, immunocytochemistry, and Western blotting. The functions of IL-33, TNF-a, and IFN-y in allergic contact dermatitis were evaluated using a murine model. RESULTS: TNF-alpha and IFN-gamma induced expression of IL-33 mRNA and protein in KERTr cells. Blockade of IL-33 attenuated swelling in the ears of the experimental mice. Similar effects were noted for blockade of TNF-alpha and IFN-gamma in these mice. CONCLUSIONS: TNF-alpha and IFN-gamma induce expression of IL-33, and IL-33 produced by keratinocytes contributes to allergic contact dermatitis. Blockade of IL-33, TNF-alpha, and IFN-gamma could represent novel and potent strategies to treat allergic contact dermatitis.

Efficacy and safety of multitarget therapy with mizoribine and tacrolimus for systemic lupus erythematosus with or without active nephritis.

The prognosis of lupus nephritis (LN) has improved since the introduction of immunosuppressant therapies, but the safety and effectiveness of treatments can also be improved. We retrospectively assessed the treatment courses of 12 patients with systemic lupus erythematosus who were treated with glucocorticoid, mizoribine (MZR) and tacrolimus. This regimen was used as initial therapy for active LN in six patients (mean glucocorticoid dose, 66.6 mg); four of these six patients also received pulse methylprednisolone therapy. The starting doses of MZR and tacrolimus were 150 and 3 mg, respectively, and they were titrated as required. Five of six patients achieved complete remission and one achieved partial remission at 6 months. Five patients who completed 12-month analysis achieved complete remission. Another six patients were given the combination regimen for treating minor flares or for steroid sparing. The mean prednisolone doses were reduced from 11.0 mg at baseline to 6.6 mg at 12 months. Six patients experienced minor adverse events, including three minor infections. One patient stopped tacrolimus because of suspected toxicity. All 12 patients were successfully treated, and none experienced severe adverse events. Multitarget therapy combining glucocorticoid, MZR and tacrolimus may have the potential to become a treatment option which is effective and safe.

Detection of parasitizing coccidia and determination of host crane species, sex and genotype by faecal DNA analysis.

In Japan, the three main crane species are the endangered red-crowned crane (Grus japonensis) inhabiting Hokkaido, the northernmost island of Japan; the vulnerable hooded crane (Grus monacha); and the vulnerable white-naped crane (Grus vipio). Both the hooded and white-naped cranes migrate in winter to Izumi in Kyushu, the southern island of Japan. In this study, we investigated the cranes and their coccidian parasites, through a targeted molecular approach using faecal DNA to develop a noninvasive method for infectious disease research. To determine the origin of noninvasively collected faecal samples, host species were identified by sequencing a region of approximately 470 bp of the mitochondrial 16S ribosomal RNA gene in the faecal DNA. Furthermore, to avoid sample redundancy, individual determination was performed by fragment analysis using microsatellite and sex-linked markers. For microsatellite genotyping, previously reported markers and markers isolated in this study were examined, and seven loci for red-crowned cranes, eight for hooded cranes and six for white-naped cranes displayed polymorphisms. A low error rate was demonstrated by comparing microsatellite data generated from faecal DNA samples with that generated from feather DNA samples, indicating a high reliability. Polymerase chain reaction-based capillary electrophoresis (PCR-CE), employing genetic markers in the second internal transcribed spacer (ITS2) of nuclear ribosomal DNA, was employed to detect crane coccidia. The sensitivity of detection of PCR-CE using faecal DNA was inferior to that with traditional microscopy; however, our results suggest that PCR-CE can depict crane coccidia diversity with higher resolution and it is a useful tool to characterize community composition of coccidia in detail.

Response to air insufflation in patients with non-erosive gastro-oesophageal reflux disease (NERD).

This study compared changes in intragastric pressure (IGP) in response to a gastric distension stimulus caused by air insufflation in 25 patients with non-erosive gastro-oesophageal reflux disease (NERD) and 34 healthy volunteers (controls). In four NERD patients, IGP responses were also measured before and after oral treatment with 10 mg rabeprazole daily for 2 weeks. The rate of increase in IGP at the start of insufflation was significantly higher in NERD patients than in controls. The time to appearance of symptoms (i.e. time to threshold) was significantly shorter in NERD patients than in controls. Direct measurement of IGP showed that visceral hypersensitivity and impaired fundal relaxation are both characteristic of NERD. Administration of rabeprazole led to a marked prolonging of time to threshold, indicating that treatment could alleviate this hypersensitivity.

Lifestyle, weight perception and change in body mass index of Japanese workers: MY Health Up Study.

OBJECTIVES: To investigate the effect of weight perception and lifestyle on body mass index (BMI) over a 2-year period. STUDY DESIGN: Longitudinal study to compare the change in BMI (kg/m(2)) according to weight perception and lifestyle at baseline. METHODS: Study subjects were 6029 men and 18,567 women aged 20-69 years who worked at a large financial firm in Japan. Subjects' weight and height were measured in 2004 and 2006. The data in 2004 were used as baseline data. Weight perception and lifestyle factors, including eating, physical exercise, hours of sleep, smoking and alcohol consumption, were determined by a self-administered questionnaire in 2004. RESULTS: The age-adjusted mean change in BMI over the 2-year period was -0.0593 among men and 0.0890 among women. In men, subjects who perceived themselves to be overweight had a reduced BMI 2 years later compared with subjects who perceived themselves to be 'just right' or underweight. Multiple regression analysis of lifestyle factors, adjusted for age and BMI at baseline, indicated that less time spent commuting, not having a hobby, not having a fixed lunch time, consumption of sweets, smoking and colleagues' smoking were associated with increased BMI among men. Fewer hours of sleep, no fixed lunch time and frequent soft drink consumption were associated with increased BMI among women. CONCLUSIONS: A perception of being overweight was associated with a decrease in BMI for Japanese male workers. Positive lifestyle factors associated with a decrease in BMI in both men and women include having a fixed lunch time and being older. These factors should therefore be highlighted in future health promotion activities in workplaces.

Expression of keratinocyte growth factor and its receptor in odontogenic keratocysts.

BACKGROUND: The mitotic activity of the epithelial cells of odontogenic keratocysts (OKCs) is greater than that of other odontogenic jaw cysts, and the mitotic activity of the epithelial cells decreases after marsupialization. Keratinocyte growth factor (KGF) interacts with its specific receptor (KGFR), and elicits the proliferation and/or differentiation of the various types of epithelial cells. The aim of this study was to investigate the expression of KGF/KGFR in OKCs before and after marsupialization. METHODS: The expression of KGF was immunohistochemically detected in the specimens of 16 OKCs and 11 dentigerous cysts before and after marsupialization. The expression of KGF mRNA was measured in the fibroblasts isolated from OKCs by real-time PCR. RESULTS: KGF was expressed in the epithelial cells and fibroblasts of 12 and seven of 16 OKC specimens, respectively. The intensity of the KGF expression in both the epithelial cells and the fibroblasts significantly decreased after marsupialization. KGFR was expressed throughout the epithelium in 15 of 16 OKC specimens, but the intensity of the KGFR expression did not change after marsupialization. The expression of KGF was detected in the epithelium of two of 11 dentigerous cyst specimens, but not in the fibroblasts before marsupialization. Real-time PCR revealed that recombinant human interleukin (IL)-1alpha increased the expression of KGF mRNA in the fibroblasts isolated from OKCs. CONCLUSION: KGF/KGFR signaling may play a crucial role in the epithelial cells of OKCs. Furthermore, the expression of KGF in the fibroblasts of OKCs is regulated by IL-1alpha.

Signaling pathways regulating IL-1alpha-induced COX-2 expression.

Interleukin-1alpha(IL-1alpha) stimulates the production of prostaglandin E(2) (PGE(2)) in odontogenic keratocyst fibroblasts. However, the signaling pathways remain obscure. In this study, we investigated IL-1alphasignaling pathways that regulate cyclooxygenase-2 (COX-2) expression in odontogenic keratocyst fibroblasts. IL-1alphaincreased the expression of COX-2 mRNA and protein, and PGE(2) secretion in the fibroblasts. IL-1alphaincreased the phosphorylation of extracellular signal-regulated protein kinase-1/2 (ERK1/2), p38 mitogen-activated protein kinase (MAPK), and c-Jun N-terminal kinase (JNK). PD-98059, SB-203580, SP-600125, and PDTC-which are inhibitors of ERK1/2, p38, JNK, and nuclear factor-kappaB (NF-kappaB), respectively-attenuated the IL-1alpha-induced COX-2 mRNA expression and activated protein kinase C PGE(2) secretion. IL-1alpha(PKC), and PKC inhibitor staurosporine inhibited IL-1alpha-induced phosphorylation of ERK1/2, p38, and JNK, and decreased IL-1alpha-induced COX-2 mRNA expression. Thus, in odontogenic keratocyst fibroblasts, IL-1alphamay stimulate COX-2 expression both through the PKC-dependent activation of ERK1/2, p38, and JNK signaling pathways, and through the NF-kappaB cascade.

Surgical results of combined pars plana vitrectomy, phacoemulsification, and intraocular lens implantation.

PURPOSE: To evaluate the results and complications of combined pars plana vitrectomy (PPV), phacoemulsification and aspiration (PEA), and intraocular lens (IOL) implantation. METHODS: A total of 117 eyes from 114 patients who had undergone PPV combined with PEA and IOL implantation were retrospectively analyzed. Combined surgery was performed for a wide variety of vitreoretinal diseases. Intraoperative and postoperative complications were also reviewed. RESULTS: The postoperative BCVA improved by 2 lines or more in 85 eyes (72.6%). Intraoperative complications consisted of retinal tears in 14 eyes (12.0%) and posterior capsular rupture in 2 eyes (1.7%). Iatrogenic retinal tears occurred more frequently in eyes with a macular hole than in eyes with any other disease (p=0.005, chi-square test). Postoperative complications consisted of posterior capsule opacification (PCO) (21 eyes), transient IOP elevation (29 eyes), vitreous hemorrhage (6 eyes), anterior chamber fibrin exudation (11 eyes), posterior iris synechia (8 eyes), neovascular glaucoma (1 eye), and recurrent retinal detachment (RD) (2 eyes). Fibrin exudation occurred more frequently in eyes with proliferative diabetic retinopathy (PDR) and RD than in eyes with any other disease (p=0.03, chi-square test). PCO occurred more frequently in eyes with PDR than in eyes with any other disease (p=0.03, chi-square test). CONCLUSIONS: The present study suggests that a high success rate can be achieved when recently improved PPV techniques are combined wi th PEA and IOL implantation. The complications that were observed following this combined treatment varied with respect to the vitreoretinal disease present prior to surgery.

Ancient DNA from pollen: a genetic record of population history in Scots pine.

Assessments of plant population dynamics in space and time have depended on dated records of fossil pollen synthesized on a subcontinental scale. Genetic analyses of extant populations have revealed spatial relationships that are indicative of past spatial dynamics, but lack an explicit timescale. Synthesis of these data requires genetic analyses from abundant dated fossil material, and this has hitherto been lacking. Fossil pollen is the most abundant material with which to fill this data gap. Here we report genetic analyses of fossil pollen retrieved from Holtjärnen postglacial lake sediment in Sweden and show that plastid DNA is recoverable from Scots Pine and Norway spruce pollen grains that are 100 and 10 000 years old. By sequencing clones from two short plastid PCR products and by using multiple controls we show that the ancient sequences were endogenous to the fossil grains. Comparison of ancient sequences and those obtained from an extant population of Scots pine establishes the first genetic link between extant and fossil samples in this species, providing genetic continuity through time. The finding of one common haplotype present in modern, 100-year old and 10 000-year old samples suggests that it may have persisted near Holtjärnen throughout the postglacial period. This retrieval of ancient DNA from pollen has major implications for plant palaeoecology in conifer species by allowing direct estimates of population dynamics in space and time.

Enlarged focal nodular hyperplasia of the liver under the influence of oral contraceptives.

Two cases of focal nodular hyperplasia of the liver in mature women treated with the oral contraceptive are described. Radiological investigations in one case revealed the typical findings of focal nodular hyperplasia with computed tomography and magnetic resonance imaging demonstrating central scar structures while spoke-wheel appearance was evident on arteriography, in the other case however findings were atypical. Routine investigations including liver function tests and alpha-fetoprotein levels were normal while hepatitis B surface antigen and hepatitis C virus antibody were negative. The lesions of these two cases enlarged significantly during the follow-up and they were therefore surgically resected. Pathological features of both resected specimens, such as hepatocellular hyperplasia, bile duct proliferation and vascular abnormalities, were compatible with focal nodular hyperplasia. It has been suggested that tumor growth may be augmented by sex hormone stimulation and therefore estrogen and progesterone receptor expressions in the resected tumors were determined by immunocytochemistry. Interestingly, stainings for both receptors were negative. In case 2, the tumor was enlarging although oral contraceptive use had been discontinued for the past 7 years. These results suggest that there is no direct relationship of focal nodular hyperplasia with oral contraceptives. The role of sex hormones in focal nodular hyperplasia of the liver merits further study.

Chloroplast DNA inversion polymorphism in populations of Abies and Tsuga.

Polymorphism for a 42-kb chloroplast DNA inversion was detected in five species of Abies and two species of Tsuga based on a sample of 1,281 individuals and both Southern hybridization and polymerase chain reaction (PCR) analyses. Two haplotypes were observed in all populations and species. The 42-kb inversion is associated with a short inverted repeat that includes trnS, psaM, and trnG. The frequencies of the two haplotypes within species were very similar among the five species of Abies This polymorphism has been maintained within populations and species in both Abies and Tsuga, probably because the mutation rate of the inversion is high. Haplotype frequencies had no geographical tendencies for any species except Abies mariesii, in which haplotype frequencies varied clinally, possibly as a result of rapid dissemination after the most recent glacial period and random genetic drift.

Molecular phylogenetic position of Japanese Abies (Pinaceae) based on chloroplast DNA sequences.

The phylogenetic positions of Japanese Abies species (A. firma, A. homolepis, A. veitchii, A. sachalinensis, and A. mariesii) were revealed based on nucleotide sequences of chloroplast DNA. First, rbcL sequences of 24 Abies species worldwide were analyzed to clarify the phylogenetic position of the Japanese Abies within this genus. While 4 of the Japanese species formed a monophyletic group with almost no sequence divergence, A. mariesii was placed into a clearly different group with North American species. Second, to clarify the relationships among the species in the monophyletic group, sequences of rbcL, matK, and six spacer regions (total ca. 5 kb) were analyzed for 12 species of the group, with A. mariesii as an outgroup. Although a unique sequence and some species-specific sequences were detected in A. mariesii and in A. firma and A. homolepis, respectively, A. veitchii and A. sachalinensis gave identical sequences for all the sequenced regions.

Clonal structure in a dwarf bamboo (Sasa senanensis) population inferred from amplified fragment length polymorphism (AFLP) fingerprints.

Amplified fragment length polymorphism (AFLP) fingerprints were used to reveal clonal structure of a dense population of dwarf bamboo, Sasa senanensis, in a 10-ha study plot at Sugadaira Montane Research Center, University of Tsukuba, Nagano, Japan. We generated AFLP fingerprints for 51 leaf samples, collected at 50 m intervals, using three selective primer pairs. A total of 135-166 fragments were detected per sample, and 22 different fingerprints were identified based on 24-83 differing fragments. Our results demonstrate that the S. senanensis population in our plot consists of at least 22 clones and that the largest single clone occurs over a distance of about 300 m. Furthermore, the clone distribution pattern implies a relationship between site quality and clonal structure.