Double-stranded RNA-exposed human keratinocytes promote Th1 responses by inducing a Type-1 polarized phenotype in dendritic cells: role of keratinocyte-derived tumor necrosis factor alpha,type I interferons,and interleukin-18

Dendritic cells play a key role in establishing the class of immune response against invading pathogens. Upon engagement with double-stranded RNA, a major bioactive constituent of many virus types, immature dendritic cells develop into type 1 immunostimulatory dendritic cells that promote Th1 responses. Immature dendritic cells reside in the epithelia and are in close contact with keratinocytes. We studied to what extent dendritic cells can also adopt a type 1 immunostimulatory dendritic cell phenotype indirectly, as a result of the interaction with keratinocytes responding to double-stranded RNA. In contrast to supernatants from keratinocytes activated by the combination of tumor necrosis factor alpha and interleukin-1beta, supernatants from keratinocytes activated by synthetic double-stranded RNA, polyriboinosinic polyribocytidylic acid, comprised tumor necrosis factor alpha and type I interferons, which induced maturation of human monocyte-derived immature dendritic cells. In addition, dendritic cells matured in the presence of these supernatants strongly biased the development of Th1 cells from naive Th cells. This bias was dependent on keratinocyte-derived interferon-alpha/beta and interleukin-18, as neutralization of both interferon-alpha/beta and interleukin-18 in the keratinocyte culture supernatant reduced the development of interferon-gamma-producing Th cells. These findings suggest that keratinocytes can contribute to the development of selective Th1/Th2 responses through the induction of maturation and functional polarization of dendritic cells, indicating a novel role for keratinocytes as initiators and regulators of cutaneous T-cell-mediated inflammation. In addition, these results support the concept that, in addition to direct interaction with pathogens, dendritic cells may also be activated and primed by pathogen indirectly, via the effect of resident tissue cells responding to pathogen.     

Tumor necrosis factor related apoptosis inducing ligand triggers apoptosis in dividing but not in differentiating human epidermal keratinocytes

Using serial analysis of gene expression we have previously identified the expression of several pro-apoptotic and anti-apoptotic genes in cultured human primary epidermal keratinocytes, including tumor necrosis factor related apoptosis inducing ligand (TRAIL). TRAIL is a potent inducer of apoptosis in transformed and tumor cell lines, but usually not in other cells. Here we present a study on the effect of TRAIL on cultured keratinocytes. It is shown that differentiated and undifferentiated keratinocytes undergo apoptosis after addition of TRAIL to the medium as determined by morphologic and biochemical criteria, such as cellular shrinkage and activation of caspases. The sensitivity for TRAIL differs greatly between undifferentiated and differentiating keratinocytes, however, with undifferentiated cells being much more susceptible to apoptosis. Commitment to terminal differentiation in the absence of TRAIL does not in itself induce apoptosis. In contrast to the promyelocytic cell line HL60, internucleosomal DNA fragmentation is not observed in keratinocytes, as assessed by flow cytometric analysis and agarose gel electrophoresis. Interestingly, the prime effector of DNA fragmentation, DNA fragmentation factor of 40 kDa (DFF40), is expressed in keratinocytes, yet internucleosomal cleavage fails to occur. Our data indicate that programmed cell death during keratinocyte differentiation is distinct from receptor-mediated apoptosis in response to a death ligand.     

Standing balance evaluation using a triaxial accelerometer

This paper presents a new inherently triaxial accelerometer-based system for determining the ability to maintain balance while standing. In this study, the accelerometer was placed at the back of the subject at the approximate height of the centre of mass. The data were processed to obtain five performance parameters. Paired t-tests indicated that the accelerometer measurements were able to distinguish between the different test conditions as well as or better than simultaneous AMTI force platform measurements (P < or = 0.05). The accelerometer system is fully portable, independent of inclination in space, low-cost and allows long term measurements of standing balance.     

1,25-dihydroxyvitamin D(3)-independent stimulatory effect of estrogen on the expression of ECaC1 in the kidney

Estrogen deficiency results in a negative Ca(2+) balance and bone loss in postmenopausal women. In addition to bone, the intestine and kidney are potential sites for estrogen action and are involved in Ca(2+) handling and regulation. The epithelial Ca(2+) channel ECaC1 (or TRPV5) is the entry channel involved in active Ca(2+) transport. Ca(2+) entry is followed by cytosolic diffusion, facilitated by calbindin-D(28K) and/or calbindin-D(9k), and active extrusion across the basolateral membrane by the Na(+)/Ca(2+)-exchanger (NCX1) and plasma membrane Ca(2+)-ATPase (PMCA1b). In this transcellular Ca(2+) transport, ECaC1 probably represents the final regulatory target for hormonal control. The aim of this study was to determine whether 17beta-estradiol (17beta-E(2)) is involved in Ca(2+) reabsorption via regulation of the expression of ECaC1. The ovariectomized rat model was used to investigate the regulation of ECaC1, at the mRNA and protein levels, by 17beta-E(2) replacement therapy. Using real-time quantitative PCR and immunohistochemical analyses, this study demonstrated that 17beta-E(2) treatment at pharmacologic doses increased renal mRNA levels of ECaC1, calbindin-D(28K), NCX1, and PMCA1b and increased the protein abundance of ECaC1. Furthermore, the involvement of 1,25-dihydroxyvitamin D(3) in the effects of 17beta-E(2) was examined in 25-hydroxyvitamin D(3)-1alpha-hydroxylase-knockout mice. Renal mRNA expression of calbindin-D(9K), calbindin-D(28K), NCX1, and PMCA1b was not significantly altered after 17beta-E(2) treatment. In contrast, ECaC1 mRNA and protein levels were both significantly upregulated. Moreover, 17beta-E(2) treatment partially restored serum Ca(2+) levels, from 1.63 +/- 0.06 to 2.03 +/- 0.12 mM. In conclusion, this study suggests that 17beta-E(2) is positively involved in renal Ca(2+) reabsorption via the upregulation of ECaC1, an effect independent of 1,25-dihydroxyvitamin D(3).     

Partial matrix excision or segmental phenolization for ingrowing toenails

OBJECTIVE: To decide whether partial nail extraction with phenolisation or with partial excision of the matrix should be the standard treatment in patients with ingrowing toenails of the hallux. DESIGN: Randomized clinical trial with 12-month follow-up evaluations performed by observers who did not know which procedure was applied. SETTING: Outpatient department of a surgical teaching hospital. PATIENTS: Fifty-eight consecutive patients with a total of 63 ingrowing toenails were randomized. INTERVENTION: Thirty-four partial matrix excisions ("matrix" group) and 29 phenolizations ("phenol" group) were performed. MAIN OUTCOME MEASURES: Recurrence rate, postoperative morbidity (pain, wound exudates, and scar discomfort), and time to complete recovery (wearing shoes, performing normal activities/work). RESULTS: Recurrences were seen after 7 procedures in the matrix group and also after 7 procedures in the phenol group, of which patients were symptomatic and required a second operation in 4 and 3 instances, respectively. None of the observed differences in wound healing, postoperative pain, and recovery were statistically significant. CONCLUSIONS: Partial matrix excision and phenolization are equally effective in treating ingrowing toenails. Because the use of the toxic agent phenol should be avoided, partial matrix excision is the preferable procedure. But in view of the high recurrence rate, there is a need for further improvement of the treatment of ingrowing toenails.     

A null mutation in the cystatin M/E gene of ichq mice causes juvenile lethality and defects in epidermal cornification

Cystatin M/E (CST6 ), a new member of the cystatin gene family, has a restricted expression pattern in humans, which is largely limited to cutaneous epithelia. Although cystatin M/E possesses two distinct biochemical properties, being a cysteine proteinase inhibitor and a substrate for transglutaminase, its physiological function is unknown. Here we report the isolation and characterization of the mouse Cst6 orthologue and the assignment of the chromosomal localization to the proximal end of mouse chromosome 19. This region corresponds to the locus of the spontaneous harlequin ichthyosis (ichq) mouse mutation, for which no causative gene has been identified so far. We found a nonsense mutation in the Cst6 gene of BALB/cJ-ichq/+ mice, which precludes the synthesis of functional protein. Immunohistochemistry confirmed the absence of cystatin M/E at the protein level in ichq/ichq mice. Mice that are homozygous for two null alleles display a hyperplastic, hyperkeratotic epidermis and abnormal hair follicles, and die between 5 and 12 days of age. In wild-type mice, cystatin M/E was found in the stratum granulosum and in the infundibulum of the hair follicle indicating that the anatomical site in the skin where cystatin M/E is normally expressed correlates with the abnormalities at the tissue level in ichq/ichq mice. Our data provide evidence that cystatin M/E is required for viability and for correct formation of cornified layers in the epidermis and hair follicles. The ichq mouse mutation may serve as a model for human type 2 harlequin ichthyosis.     

Lack of isoprenoid products raises ex vivo interleukin-1beta secretion in hyperimmunoglobulinemia D and periodic fever syndrome

OBJECTIVE: To investigate whether the increased interleukin-1beta (IL-1beta) secretion in hyperimmunoglobulinemia D and periodic fever syndrome is due to the accumulation of mevalonate kinase (MK), the substrate of the deficient enzyme, or the lack of its products, the isoprenoid compounds. METHODS: The effects of lovastatin and farnesol (FOH), geranylgeraniol (GGOH), and mevalonate on peripheral blood mononuclear cells (PBMCs) from 8 patients with MK deficiency and from 13 controls were studied. Lovastatin inhibits isoprenoid biosynthesis by reducing the production of mevalonate. FOH and GGOH restore isoprenoid biosynthesis downstream from MK. Culture supernatants were collected for cytokine analysis 48 hours after stimulation with monoclonal antibodies against CD2 + CD28. RESULTS: Lovastatin induced a 15-fold rise in IL-1beta secretion by normal anti-CD2 + CD28-stimulated cells (P < 0.001). This effect could be countered by mevalonate and, to a lesser extent, by FOH and GGOH. In the absence of lovastatin, mevalonate did not change IL-1beta secretion. Stimulated MK-deficient cells secreted 9-fold more IL-1beta than control PBMCs (P < 0.005), rising 2.4-fold in the presence of lovastatin. The effect of lovastatin on IL-1beta secretion was reduced by mevalonate, FOH, and GGOH. Isoprenoid biosynthesis in PBMCs from patients was impaired due to the endogenous MK deficiency. Bypassing this defect with FOH, in the absence of lovastatin, led to a 62% reduction (P < 0.02) in IL-1beta secretion by these cells. CONCLUSION: In this model, shortage of isoprenoid end products contributes to increased IL-1beta secretion by MK-deficient PBMCs, whereas excess mevalonate does not.     

Visual Acuity Development After the Implantation of Unilateral Intraocular Lenses in Infants and Young Children

PURPOSE: Intraocular lenses (IOLs) are now being implanted in infants and children with unilateral cataracts. However, there are no prospective data on the development of visual acuity after implantation. The aim of the present study was to prospectively assess the development of acuity in infants and preschool children who received IOLs or aphakic contacts lenses (CLs) after the extraction of a unilateral cataract. METHODS: Visual acuity was assessed using Teller Acuity Cards and/or crowded HOTV tests at target ages of 6 months, 1, 2, 3, and 4 years. RESULTS: Infants who received a primary IOL after extraction of dense congenital unilateral cataract (n = 5) showed improvement from an initially low mean visual acuity of 20/170 at 6 months to 20/85 at 12 months and 20/54 at 4 years. Visual acuity in the IOL group was similar to that of children who had good-to-excellent compliance with CL wear (n = 36; 4-year visual acuity 20/50) and better than that of children who had moderate-to-poor compliance (n = 11; 4-year visual acuity 20/135). Children who received IOLs after extraction of developmental unilateral cataracts by 6 months (n = 4; 4-year visual acuity 20/55) had visual acuity development similar to those treated with CLs (n = 5; 4-year visual acuity 20/55). Children who received IOLs after extraction of developmental unilateral cataracts after 1 year of age (n = 18) had better visual acuity than children those treated with CLs (n = 4) at 4 years of age (20/40 vs. 20/135). CONCLUSION: IOLs and aphakic CLs support similar visual acuity development after surgery for a unilateral cataract. IOLs may support better visual acuity development when compliance with CL wear is moderate to poor or when a cataract is extracted after 1 year of age.     

A study on symptom profiles of late-life depression: the influence of vascular, degenerative and inflammatory risk-indicators

BACKGROUND: If specific symptom profiles of depressive disorders in the elderly are related to a specific etiology, this could have implications for everyday clinical practice. We hypothesized that a "motivational" profile, with symptoms such as psychomotor change, loss of interest and loss of energy, could clinically separate patients with predominantly vascular or degenerative risk indicators from patients with inflammatory risk indicators. METHODS: A total of 4051 subjects participated in a study on mental health problems in community-dwelling elderly. Information on psychiatric symptoms, demographic and medical status, previous history and family history was obtained. We distinguished three subgroups according to predominant somatic risk-indicators; vascular, degenerative and inflammatory groups. RESULTS: Motivational symptoms were associated with vascular or degenerative risk-indicators for depression; psychomotor change with both indicators; loss of energy with vascular, though also with the inflammatory indicator, and thinking/concentration disturbance with the degenerative indicator. The so-called mood symptoms of depression, especially thoughts of death, were more strongly related with the inflammatory risk-indicator. Melancholic symptoms like appetite and sleep disturbances were more strongly associated with the inflammatory risk-indicator. LIMITATIONS: Etiological classification was not confirmed by additional investigations such as laboratory findings or MRI brain scans. CONCLUSION: This study showed that in patients with a late-life depression specific symptoms of the depressive disorder may reflect the predominant underlying pathogenic mechanism.