Ichthyosis with confetti caused by new and recurrent mutations in KRT10 associated with varying degrees of keratin 10 mis-localization.

BACKGROUND: Ichthyosis with confetti (IWC) is an extremely rare autosomal-dominant genodermatosis characterized by erythroderma with numerous confetti-like pale spots. IWC is caused by mutations in KRT10 (IWC-I) or KRT1 (IWC-II) which affect their tail domains. In IWC-I, the mutations lead to replacement of glycine/serine-rich keratin 10 (K10) tail with arginine- or alanine-rich frameshift motifs, causing K10 mis-localization which might trigger loss of the mutant KRT10 allele via mitotic recombination, leading to genetic reversion. OBJECTIVE: To investigate mutations in five IWC-I patients and their functional consequences. METHODS: We performed Sanger sequencing of KRT1 and KRT10 in peripheral blood samples of five patients, with highly polymorphic KRT10 SNPs genotyped to confirm loss-of-heterozygosity in the epidermis of pale spots. K10 expression pattern was examined in both patient skin biopsies and HaCaT cells overexpressing mutant KRT10-enhanced green fluorescence protein fusion. RESULTS: Four novel and one recurrent KRT10 mutations were identified in patient peripheral blood samples but not in the corresponding pale spot epidermis. Two of the mutations, c.1696_1699dupCACA and c.1676dupG, affected residues close to K10 carboxyl terminus and encoded only 3 and 6 arginine residues, which were far fewer than reported previously. Interestingly, imaging analyses for K10 in HaCaT cells overexpressing either of these two mutations and in the corresponding patients' affected skin, showed a remarkably lower level of K10 mis-localization compared to that of other mutations reported in this study. CONCLUSIONS: Our findings suggest that the number of arginine residues in the mutant tail may correlate with the level of K10 mis-localization in IWC-I keratinocytes. These results expand the genotypic and phenotypic spectrum of IWC-I.

The Major Orphan Forms of Ichthyosis Are Characterized by Systemic T-Cell Activation and Th-17/Tc-17/Th-22/Tc-22 Polarization in Blood.

The ichthyoses are rare skin disorders with immune and barrier aberrations. Identifying blood phenotypes may advance targeted therapeutics. We aimed to compare frequencies of skin homing/cutaneous lymphocyte antigen (+) versus systemic/cutaneous lymphocyte antigen (-) "polar" CD4+/CD8+ and activated T-cell subsets in ichthyosis versus atopic dermatitis, psoriasis, and control blood, with appropriate clinical correlations. Flow cytometry was used to measure IFN-γ, IL-13, IL-9, IL-17, and IL-22 cytokines in CD4+/CD8+ T cells, with inducible co-stimulator molecule and HLA-DR defining mid- and long-term T-cell activation, respectively. We compared peripheral blood from 47 patients with ichthyosis (congenital ichthyosiform erythroderma, lamellar ichthyosis, epidermolytic ichthyosis, and Netherton syndrome) with 43 patients with atopic dermatitis and 24 patients with psoriasis and 59 age-matched controls. Clinical measures included the ichthyosis severity score, with subsets for erythema and scaling, transepidermal water loss, and pruritus. All ichthyoses had excessive inducible co-stimulator molecule activation (P < 0.001), particularly epidermolytic ichthyosis. Significantly elevated IL-17- (P < 0.05) and IL-22-producing (P < 0.01) T cells characterized ichthyoses, mainly Netherton syndrome and congenital ichthyosiform erythroderma (P < 0.05). Increased T helper 2/cytotoxic T cell 2/T helper 9 (P < 0.05) and similar IFN-γ frequencies (P > 0.1) versus controls were also noted. IL-17/IL-22-producing cells clustered with clinical measures, whereas IFN-γ clustered with age. Our data show peripheral blood IL-17/IL-22 activation across the ichthyoses, correlating with clinical measures. Targeted therapies should dissect the relative contribution of polar cytokines to disease pathogenesis.

Identification of a novel mutation in ZAP70 and prenatal diagnosis in a Turkish family with severe combined immunodeficiency disorder.

Protein tyrosine kinases (PTKs) play an important role in T cell development and activation. In vitro and in vivo defects, resulting in variable deficiencies in thymic development and in T cell antigen receptor (TCR) signal transduction, in PTKs have been shown. ZAP70, one of those PTKs, is a 70-kDa tyrosine phosphoprotein and associates with the ζ chain and undergoes tyrosine phosphorylation following TCR stimulation. It is expressed in T and natural killer (NK) cells. Several mutations were shown to lead to an autosomal recessive form of severe combined immunodeficiency disease (SCID). Here, we present a family with a novel mutation in ZAP70. The proband, the second child of the first cousin parents of Turkish origin, was diagnosed with SCID having R514C mutation on homozygous state. She had decreased CD8(+) T and natural killer cells, normal CD4(+) T cells, high serum Ig E level, perivascular dermatitis and ichthyosis. This article presents clinical features of a novel mutation on ZAP70 and the first prenatal molecular diagnosis of ZAP70 deficiency. Different mutations in ZAP70 and related phenotypes reported in the literature are also discussed.

[The evaluation of nitrogen metabolism and the reactivity of the organism in patients with dry skin].

It is known that in norm horny layer of the epidermis is able to retain water due to the presence of hygroscopic substances inside corneocytes in the form of so-called natural moisturizing factors (NMF), consisting of free amino acids and their derivatives, which are formed during the decay of filaggrin as well as lactic acid, urea, sugars, and intercellular lipid membranes, creating a barrier that prevents transepidermal water loss. At the same time, the results of recent studies have shown that urea--a kind of natural antioxidant that protects tissues from the accumulation of aggressive forms of oxygen. It is able to stabilize the lysosomal membranes, thus preventing autolysis of cells. The ability of urea at low concentrations to modify the reactivity of functional groups of proteins leads to conformational changes of immunoglobulin, which has an inhibitory effect on the immune system, including the diminishing impact on the development of reaginic type reactions. Urea has anti-inflammatory, hyposensitizing, and antioxidant effect. Based on the above the aim of this study was to determine the content of urea and some indicators of cellular and humoral immunity in case of chronic dermatoses, accompanied by dryness of the skin. Indicators of nitrogen metabolism of blood serum (urea. ammonia), some parameters of cellular and humoral immunity were studied in 27 patients, who according to nosological units were distributed as follows: atopic dermatitis (12), psoriasis (7), xerosis (8). In the study of the concentration of urea in the blood, and some indicators of cellular immunity, as well as the content of immunoglobulin E in the blood of our patients a decrease in the number of T--lymphocytes, mainly due to T--suppressor and raising the level of immunoglobulin E have been revealed. Specific patterns of changes in these parameters, depending on nosological unit, severity of disease and degree of dryness of the skin have also been observed.

A syndrome of retinitis pigmentosa, congenital ichthyosis, hypergonadotropic hypogonadism, small stature, mental retardation, cranial dysmorphism, and abnormal electroencephalogram.

BACKGROUND: Retinal dystrophy and ichthyosis occur together in patients with the well-characterized disorders of Refsum disease and Sjögren-Larsson syndrome. Rud syndrome formerly was considered a genetically heterogeneous but distinct clinical entity with the manifestations of icthyosis, hypogonadism, small stature, mental retardation, epilepsy, and, infrequently, retinitis pigmentosa. Although there are at least 55 case reports of Rud syndrome in the medical literature, the existence of such a syndrome has recently been dismissed based on a new understanding of the ichthyoses. Most case reports previously reported as Rud syndrome can now be reassigned under a contemporary ichthyosis classification that does not include Rud syndrome as a distinct entity. METHODS: Two unrelated women with a disorder showing retinitis pigmentosa, congenital ichthyosis, hypergonadotropic hypogonadism, small stature, mental retardation, cranial dysmorphism, and abnormal electroencephalograms underwent a comprehensive workup. The ocular and systemic findings are compared with those previously described for retinal dystrophy and ichthyosis disorders. RESULTS: These cases were found to be clearly distinct from Refsum disease, Sjögren-Larsson syndrome, and any of the other ichthyosis disorders that have been suggested as a replacement for Rud syndrome. CONCLUSION: The association of retinitis pigmentosa, congenital ichthyosis, hypergonadotropic hypogonadism, small stature, mental retardation, cranial dysmorphism, and abnormal electroencephalogram may represent a distinct syndrome previously considered a subset of the now defunct Rud syndrome.

Serum lipids in children with xeroderma from an inland district of Sri Lanka.

Children living on plantations in inland districts of the southeastern part of Sri Lanka frequently develop a skin condition on the legs described as mosaic skin or xeroderma. This condition is characterized by atrophic, dry, shining and scaly skin. The etiology is unknown. A food frequency survey indicated a low energy intake, a diet with a low fat content, and anthropometric data have shown a high prevalence of malnutrition within this group. The skin condition brought attention to a possible deficiency of essential nutrients, especially essential fatty acids. In order to investigate the possible association with a deficiency of essential fatty acids, blood samples were collected from both children having signs of xeroderma and controls. The total amount of phospholipids was low, but the fatty acid profile of this lipid class was similar to the controls. A vitamin A deficiency was indicated by low levels of its transport proteins. A multifactorial etiology where vitamin A deficiency may play a role is discussed.

Serum parathyroid hormone level is elevated in some patients with disorders of keratinization.

BACKGROUND AND DESIGN: After the chance of observation of an elevated parathyroid hormone (PTH) value in a patient with pityriasis rubra pilaris, the serum PTH level was measured in the next 14 patients seen with disorders of keratinization. Calcium metabolism in three affected patients was then studied in depth. RESULTS: Five of 15 patients had twofold or greater elevations in serum PTH values. The patients had four different disorders of keratinization: bullous congenital ichthyosiform erythroderma (two patients); lamellar ichthyosis (one patient); pityriasis rubra pilaris (one patient); and ichthyosis linearis circumflexa (one patient). At least one other patient with each diagnosis had normal PTH values. Two of three patients who were studied further had clear evidence of increased, biologically active PTH, consistent with secondary hyperparathyroidism. An elevated PTH level spontaneously became normal in one patient, and in a second patient it became normal with a high-calcium diet. CONCLUSIONS: These data provide the first indication that patients with various disorders of keratinization have an increased risk for secondary hyperparathyroidism. The exact prevalence, origin, and physiologic significance of this finding remain to be elucidated.

Neutral lipid storage disease with ichthyosis: serum apolipoprotein levels and cholesterol metabolism in monocyte-derived macrophages.

Neutral lipid storage disease with ichthyosis (NLSDI) is an inherited metabolic disorder characterized by accumulation of neutral lipids, in a wide variety of cells, by a still unknown mechanism. Previous studies have shown normal cholesterol content in NLSDI granulocytes, fibroblasts and skin cells. Monocyte-derived macrophages possess an additional pathway of cholesterol uptake, which is not shared by these cells and which is not regulated by intracellular cholesterol levels. This pathway is thought to play a rôle in the process of atherosclerosis. Three NLSDI patients were studied. The serum levels of triglycerides, cholesterol, high-density lipoprotein cholesterol, and apolipoproteins A-I and B were within normal limits in all three patients. The intracellular levels of free and esterified cholesterol were measured in the monocyte-derived macrophages of one patient and found to be normal, while the triglyceride concentrations were twice as high as normal. The cholesterol esterification rates, which serve as a sensitive indicator of intracellular changes in cholesteryl ester levels, were normal in the monocyte-derived macrophages of all three patients. These findings provide further evidence that cholesterol metabolism is not disturbed in NLSDI, and it may be inferred that in this respect these patients are not at increased risk for atherosclerosis.

Essential-fatty-acid metabolites in plasma phospholipids in patients with ichthyosis vulgaris, acne vulgaris and psoriasis.

The concentration of essential fatty acids (EFAs) and their metabolites in plasma phospholipids were measured by gas chromatography in normal individuals, and in patients with ichthyosis vulgaris, acne vulgaris or psoriasis. In all three patient groups, concentrations of arachidonic acid (20:4 omega 6) and docosapentaenoic acid (22:5 omega 6) were significantly below those in controls, suggesting that these abnormalities may occur in many skin diseases. Concentrations of dihomogammalinolenic acid (20:3 omega 6) were low in ichthyosis, normal in acne and elevated in psoriasis. Thus ichthyosis, acne and psoriasis each had a characteristic pattern of EFA metabolites.

Search for consanguinity within and among families of patients with trichothiodystrophy associated with xeroderma pigmentosum.

The association of two rare hereditary disorders, trichothiodystrophy (TTD) and xeroderma pigmentosum (XP), was found in four patients from three families, apparently unrelated but living in the same geographical area. In order to test the hypothesis of a common ancestor, consanguinity within and among the families was checked using three different approaches: reconstruction of genealogical trees, typing of blood markers, and surname analysis. The results of the three types of analyses strengthen the hypothesis that, in at least two out of the three families, the genetic defect determining the TTD/XP phenotype is identical by descent, as a consequence of remote inbreeding. This implies that if two mutations are responsible for the two diseases they are at linked loci or affect the same gene.

Diagnosis of recessive X-linked ichthyosis: quantitative HPLC/mass spectrometric analysis of plasma for cholesterol sulfate.

A specific, accurate liquid-chromatographic/mass-spectrometric (HPLC/MS) method for measurement of cholesterol sulfate in plasma from normal individuals and patients with recessive X-linked ichthyosis (RXLI) is described. The method is superior to previously described techniques because it measures the analyte intact rather than after hydrolysis. Traces of free cholesterol in the sample analyzed do not add to the measured result. We used either [13C2]cholesterol sulfate or [2H6]cholesterol sulfate as internal standards, which we add to plasma before extraction. Use of such standards makes quantitative extraction unimportant. We use a single solid-phase extraction (SPE) C18 cartridge for plasma extraction. After the cartridge is washed with methanol/ammonium acetate solutions, the fraction containing the steroid sulfates is eluted with methanol, evaporated, and subjected to HPLC/MS analysis, wherein the molecular anions of analyte and internal standards are monitored. The peak ratio gives the cholesterol sulfate concentration directly. Using this method, we have diagnosed 24 patients with RXLI. Their concentration of cholesterol sulfate ranged between 41.7 and 185.3 mumol/L (mean 93.85, SD 31.2 mumol/L). In normal individuals (n = 9) the mean cholesterol sulfate concentration was 2.77 mumol/L (SD 0.62, range 2.05-3.95 mumol/L). The instrumentation required is complex, but the assay is simple: sample preparation takes about 30 min; mass spectrometry, 10 min. About 0.1 mL of plasma is required for cholesterol sulfate measurement in RXLI patients and 0.5-1 mL in normal individuals.

Nyctalopia and conjunctival xerosis indicating vitamin A deficiency in cystic fibrosis.

Thirty-one Cystic Fibrosis patients were investigated for clinical and biochemical evidence of Vitamin A deficiency. All had been prescribed oral pancreatic enzyme replacements and twice the recommended daily requirement of Vitamin A (5000IU). None were aware of any ocular symptoms, but 3 out of 31 (10 per cent) were found to have frank conjunctival xerosis and six (19 per cent) to have abnormal dark adaptation. There was no correlation between the above findings and abnormal liver function or clinical disease severity. All patients with cystic fibrosis should have regular Vitamin A estimations with ophthalmological assessment if serum levels fall below 30 microgram/dl.

Simplified determination of serum cholesterol sulfate by gas-liquid chromatography combined with cyclohexylsilane-bonded phase column purification.

A new gas-liquid chromatographic (GLC) determination of cholesterol sulfate (CS) and dehydroepiandrosterone sulfate (DHEAS) for a biochemical diagnosis of recessive X-linked ichthyosis (RXLI) is described. Although the GLC method for determination of CS is known to be more sensitive than the thin layer chromatographic (TLC) method, the former method has not been widely employed because of its complicated pre-purification steps. The present method allows us to measure the serum levels of CS and DHEAS without tedious purification steps such as multiple conventional column chromatography and preoperative thin layer chromatography. Sulfated steroids are rapidly purified with a commercially available mini disposable cyclohexylsilane-bonded phase (CH) column, CH BOND ELUT, and the purified steroids after desulfation are converted to water-resistant tert-butyldimethylsilyl ether derivative for the GLC analysis on dual 2m glass columns packed with 2% XE-60 on Chromosorb W. By the present method, serum CS concentrations in RXLI patients were shown to be about 10 times higher than those in patients with ichthyosis vulgaris, carriers of RXLI, and healthy subjects. This method is more suitable not only for a biochemical diagnosis of RXLI but also for studies on the metabolism of sulfated steroids than the previous time-consuming GLC methods.

X-linked ichthyosis and ichthyosis vulgaris: comparison of their clinical features based on biochemical analysis.

Thirty cases of X-linked ichthyosis (XLI) and 32 cases of ichthyosis vulgaris (IV) were diagnosed by measuring the steroid sulphatase activity of peripheral blood lymphocytes or the electrophoretic mobility of serum LDL or both. The clinical features of the two conditions were then compared. In both diseases 60-66% of patients had a family history of the condition. Ichthyosis was noted at birth or immediately afterwards in 59% of the patients with XLI while it appeared in infancy in 68% of those with IV. Scales were mostly large and brown or dark brown in patients with XLI, while the majority of patients with IV had small brown or light brown scales. The distribution of the ichthyotic lesions differed in the two types of ichthyosis. On the trunk, the abdomen was more severely involved than the back in 63% of the cases with the XLI, whereas the back was more scaly than the abdomen in 44% of those with IV. On the extremities, the extensor surface was more severely affected than the flexor surface in both types. X-linked ichthyosis was characterized by the presence of lesions in the pre-auricular area, which were found in 93% of the cases with XLI, while only 17% of the IV patients had ichthyotic lesions at this site. Involvement of the preauricular area could be an important clinical feature for distinguishing XLI from IV.