Plant mitochondrial RNA editing factors can perform targeted C-to-U editing of nuclear transcripts in human cells.

RNA editing processes are strikingly different in animals and plants. Up to thousands of specific cytidines are converted into uridines in plant chloroplasts and mitochondria whereas up to millions of adenosines are converted into inosines in animal nucleo-cytosolic RNAs. It is unknown whether these two different RNA editing machineries are mutually incompatible. RNA-binding pentatricopeptide repeat (PPR) proteins are the key factors of plant organelle cytidine-to-uridine RNA editing. The complete absence of PPR mediated editing of cytosolic RNAs might be due to a yet unknown barrier that prevents its activity in the cytosol. Here, we transferred two plant mitochondrial PPR-type editing factors into human cell lines to explore whether they could operate in the nucleo-cytosolic environment. PPR56 and PPR65 not only faithfully edited their native, co-transcribed targets but also different sets of off-targets in the human background transcriptome. More than 900 of such off-targets with editing efficiencies up to 91%, largely explained by known PPR-RNA binding properties, were identified for PPR56. Engineering two crucial amino acid positions in its PPR array led to predictable shifts in target recognition. We conclude that plant PPR editing factors can operate in the entirely different genetic environment of the human nucleo-cytosol and can be intentionally re-engineered towards new targets.

The dual-targeted RNA editing factor AEF1 is universally conserved among angiosperms and reveals only minor adaptations upon loss of its chloroplast or its mitochondrial target.

KEY MESSAGE: Upon loss of either its chloroplast or mitochondrial target, a uniquely dual-targeted factor for C-to-U RNA editing in angiosperms reveals low evidence for improved molecular adaptation to its remaining target. RNA-binding pentatricopeptide repeat (PPR) proteins specifically recognize target sites for C-to-U RNA editing in the transcriptomes of plant chloroplasts and mitochondria. Among more than 80 PPR-type editing factors that have meantime been characterized, AEF1 (or MPR25) is a special case given its dual targeting to both organelles and addressing an essential mitochondrial (nad5eU1580SL) and an essential chloroplast (atpFeU92SL) RNA editing site in parallel in Arabidopsis. Here, we explored the angiosperm-wide conservation of AEF1 and its two organelle targets. Despite numerous independent losses of the chloroplast editing site by C-to-T conversion and at least four such conversions at the mitochondrial target site in other taxa, AEF1 remains consistently conserved in more than 120 sampled angiosperm genomes. Not a single case of simultaneous loss of the chloroplast and mitochondrial editing target or of AEF1 disintegration or loss could be identified, contrasting previous findings for editing factors targeted to only one organelle. Like in most RNA editing factors, the PPR array of AEF1 reveals potential for conceptually "improved fits" to its targets according to the current PPR-RNA binding code. Surprisingly, we observe only minor evidence for adaptation to the mitochondrial target also after deep losses of the chloroplast target among Asterales, Caryophyllales and Poales or, vice versa, for the remaining chloroplast target after a deep loss of the mitochondrial target among Malvales. The evolutionary observations support the notion that PPR-RNA mismatches may be essential for proper function of editing factors.

Generating Attention, Inhibition, and Memory: A Pilot Randomized Trial for Preschoolers With Executive Functioning Deficits.

This goal of this study was to assess the initial feasibility and efficacy of a play-based intervention targeting executive functions (EF) and parent-child relationships in preschoolers compared with an active control group. Preschoolers with EF deficits (M age = 3.7 ± 0.47, predominantly White boys) and their parents were randomized to intervention (n = 36) or active control (n = 32) conditions. Child performance on EF tasks, parent and masked teacher ratings of EF and behavior, and masked clinician ratings of severity were collected at baseline and at 3 and 6 months postbaseline. Partial eta-squared effect sizes at .02 or higher comparing performance across the two groups was considered evidence of meaningful, albeit small, intervention effects. Intervention effects were observed for parent ratings of inattention, hyperactivity/impulsivity, and number/severity of problems experienced in various home situations, teacher ratings of severity of problems in various school situations, parent and teacher ratings of overall impairment, and clinician ratings of impairment. Intervention effects for functional improvements were maintained at the 6-month follow-up. No effect of the intervention was observed on the objective EF measures, although parent ratings of emotional control were improved for children in the intervention group. An intervention utilizing play-based activities targeting EF, when administered in a structured way by parents, is a promising approach for improving behavior in preschoolers with self-regulation deficits. More work is needed to investigate potential impact on EF and to disentangle mechanisms of action. It may be that the intervention's focus on the structure and quality of parent-child interactions is a mediator of outcomes, rather than improved EFs.

Eponyms and clinical entities from the land of Israel.

Seven eponyms have been pioneered by dermatologists in Israel: Brenner's sign, Chanarin-Dorfman syndrome, granulated sweetener packet sign, isopathic phenomenon of Sagher, lanolin paradox, Nakar-Ingber disease, and Wolf's isotopic response. In addition, there are three id reactions described by Israeli dermatologists: leishmanid, pediculid, and scabid. There is also the acronym PEMPHIGUS, which stands for the causative reasons for pemphigus. We celebrate these eponyms and clinical entities, which reflect the impressive progress made by dermatologists in Israel during the past century who have helped to build an academic, vibrant, and dynamic specialty in the Holy Land.

In vitro release of interferon-gamma from peripheral blood lymphocytes in cutaneous adverse drug reactions.

BACKGROUND: Cutaneous drug reactions are common but diagnostically challenging due to phenotypic heterogeneity and simultaneous exposure to multiple drugs. These limitations prompted the development of diagnostic tests. AIMS: To evaluate the performance of an in vitro assay measuring interferon-gamma release from patients' lymphocytes in the presence of causative drugs for the diagnosis of drug reactions. METHODS: Mononuclear cells derived from patients were incubated with and without suspected drugs, and increment of interferon-gamma levels was measured by ELISA. We performed a telephonic survey to evaluate the effect of stopping the drugs incriminated by the assay on cutaneous manifestations. RESULTS: We assessed 272 patients who used 1035 medications. When assessed against the questionnaire data collected at least 6 months after stopping the causative drug, sensitivity was found to be 83.61% and specificity 92.67%. Likelihood ratio for a positive test is 11.40 and for a negative test 0.18. Positive predictive value is 75.37% and negative predictive value is 95.47%. The test was found to perform significantly better in females and in older patients. CONCLUSIONS: Interferon-gamma release test is a useful adjunct tool in the diagnosis of cutaneous drug reactions.

Effect of Solar Radiation on Skin Microbiome: Study of Two Populations.

Here, we examined the skin microbiome of two groups of healthy volunteers living on the Mediterranean coast with different exposures to sun radiation. One group, exposed to the sun in the summer, was compared with a group covered with clothing throughout the year. The seasonal effects on the skin microbiome of three body sites were determined before and after summer. Surprisingly, at the phyla level, there were no significant differences in microbiome diversity between the groups. Furthermore, within each group, there were no significant seasonal differences in high-abundance species at any of the sampling sites. These results suggest that the skin microbiome, developed over years, remains stable even after several months of exposure to summer weather, direct sunlight and humidity. However, in the group exposed to the sun during the summer months, there were significant differences in low-abundance species in sun-exposed areas of the skin (the inner and outer arm). These subtle changes in low-abundance species are interesting, and their effect on skin physiology should be studied further.

Differences in cytokine levels in melanoma patients with and without redness (Brenner sign).

BACKGROUND: In view of several studies highlighting an observation of an erythematous eruption in the vicinity of or distant from the lesion in melanoma patients (The Brenner sign), this study sought to assess whether this phenomenon might be related to the blood level of cytokines IL-6 and IL-8. PATIENTS AND METHODS: Sera specimens obtained from 27 patients with melanoma, of which 15 had erythematous eruptions and 12 did not, were studied by immunohistochemistry for the expression of IL-6 and IL-8. RESULTS: IL-6 was detected in all melanoma patients in both groups. The mean level of IL-6 in the redness group (2.41 pg/L) was significantly higher than in the group without redness (1.25 pg/L). IL-8 was detected in all 27 melanoma patients in the two groups. The serum level was less than 5 pg/L in only 1 patient (6.7%) in the redness group, and in 6 patients (50%) in the group without redness, a statistically significant difference. CONCLUSION: The Brenner sign appears to reflect a more advanced disease and herald a poor prognosis according to its correlation with the IL-8 and IL-6 blood level. However, in view of the biphasic effect of IL-8 level on tumor progression, and IL-6's ability to inhibit early stage melanoma, redness in melanoma patients could be a sign of a better prognosis of the melanoma.

Eczema craquelé with purpura: a sign of internal malignancy or malabsorption syndrome?

We describe a patient with eczema craquelé associated with adenocarcinoma of the pancreas. Following a Whipple operation, zinc malabsorption was ruled out as the cause based on the distribution of the dermatitis, alkaline phosphatase levels within reference range, and response to pancreatic enzyme replacement therapy. Because the patient's skin changes appeared following removal of the malignancy and resolved shortly after the second course of pancreatic enzyme replacement therapy, fat malabsorption probably was responsible for the skin changes rather than a direct paraneoplastic expression of the cancer. The occurrence of purpura with eczema craquele has not been previously described in the literature. These findings raise the possibility that generalized eczema craquele is a diagnostic clue to a malignancy and argue for adding it to the list of cutaneous paraneoplastic syndromes.

Multifarious Evolutionary Pathways of a Nuclear RNA Editing Factor: Disjunctions in Coevolution of DOT4 and Its Chloroplast Target rpoC1eU488SL.

Nuclear-encoded pentatricopeptide repeat (PPR) proteins are site-specific factors for C-to-U RNA editing in plant organelles coevolving with their targets. Losing an editing target by C-to-T conversion allows for eventual loss of its editing factor, as recently confirmed for editing factors CLB19, CRR28, and RARE1 targeting ancient chloroplast editing sites in flowering plants. Here, we report on alternative evolutionary pathways for DOT4 addressing rpoC1eU488SL, a chloroplast editing site in the RNA polymerase ß' subunit mRNA. Upon loss of rpoC1eU488SL by C-to-T conversion, DOT4 got lost multiple times independently in angiosperm evolution with intermediate states of DOT4 orthologs in various stages of degeneration. Surprisingly, we now also observe degeneration and loss of DOT4 despite retention of a C in the editing position (in Carica, Coffea, Vicia, and Spirodela). We find that the cytidine remains unedited, proving that DOT4 was not replaced by another editing factor. Yet another pathway of DOT4 evolution is observed among the Poaceae. Although the rpoC1eU488SL edit has been lost through C-to-T conversion, DOT4 orthologs not only remain conserved but also have their array of PPRs extended by six additional repeats. Here, the loss of the ancient target has likely allowed DOT4 to adapt for a new function. We suggest rps3 antisense transcripts as previously demonstrated in barley (Hordeum vulgare) arising from promotor sequences newly emerging in the rpl16 intron of Poaceae as a new candidate target for the extended PPR stretch of DOT4. Altogether, DOT4 and its target show more flexible pathways for evolution than the previously explored editing factors CLB19, CRR28, and RARE1. Certain plant clades (e.g., Amaranthus, Vaccinium, Carica, the Poaceae, Fabales, and Caryophyllales) show pronounced dynamics in the evolution of editing sites and corresponding factors.

Unilateral spiny hyperkeratosis: case report and review of the literature.

We describe the first case of unilateral spiny hyperkeratosis (SH) of the left hand, review the literature and discuss possible patho- mechanisms. SH can be sporadic or familial, often appearing in healthy individuals. However, there is an association with various malignancies in a significant number of the sporadic cases. Although there is no satisfactory explanation of this association, we agree with previous authors that a patient with SH appearing in adult life should be evaluated and followed for the presence of malignancy. Other patients with SH may suffer from a variety of nonmalignant diseases, which may be coincidental or causally related. SH is not a premalignant lesion of the skin and should not be confused with porokeratosis which has a malignant potential. Except for excision of individual lesions, there is no permanent cure.

In vitro interferon-gamma release test in patients with drug-induced pemphigus.

BACKGROUND: Drug-specific CD8+ TH1 lymphocytes have been found in the peripheral blood and involved skin of patients with drug-induced bullous exanthems. OBJECTIVES: To determine whether the interferon-gamma release test can identify culprit drugs in pemphigus patients. METHODS: Clinical and laboratory workup for pemphigus was performed in 14 pemphigus vulgaris patients who had been exposed to drugs, and the IFN-gamma release test was conducted on their lymphocytes from heparinized venous blood cultured with medium, phytohemagglutinin and one of 32 drugs, or medium and phytohemagglutinin alone. RESULTS: Ten of the patients and 13 of the 32 drugs exhibited a positive response to the test. Eight of the 10 patients with positive IFN-gamma test results had a less severe course of the disease, with fast reduction in steroid dosage. CONCLUSIONS: The findings demonstrate both the ability of the IFN-gamma release test to identify drugs that can induce pemphigus, and its usefulness in the diagnostic workup of pemphigus patients.

Expression of vinculin in autoimmune cutaneous diseases.

BACKGROUND: Reports have appeared that sera of patients with systemic autoimmune disorders have demonstrated autoantibodies to vinculin. OBJECTIVE: To determine the presence and distribution of vinculin in the skin of patients with cutaneous autoimmune disorders. METHODS: Semiquantitative immunohistochemistry investigations for presence of vinculin were conducted on skin biopsy specimens from patients with pemphigus vulgaris (PV), bullous pemphigoid (BP), and various collagen vascular diseases, and from healthy controls. RESULTS: Results of staining for vinculin were positive in 2 of 7 PV patients, 6 of 9 BP patients, and all 6 cutaneous autoimmune patients. Staining results were negative in all controls. Strong immunostaining to vinculin was found in 3 of 6 vinculin-positive BP patients and 5 of 6 vinculin-positive cutaneous autoimmune patients. CONCLUSIONS: The expression and distribution of vinculin are accentuated in patients with various skin autoimmune diseases and appear to be stronger in diseases involving the basement membrane, where it is thought to be relatively more important than in other skin disorders.

Are sluggish cognitive tempo symptoms associated with executive functioning in preschoolers?

The aim of this study is to investigate whether sluggish cognitive tempo (SCT) symptoms are associated with neurocognitive task performance and ratings of real-world executive functioning (EF) in preschoolers at risk for attention-deficit/hyperactivity disorder (ADHD). The associations between parent- and teacher-rated SCT symptoms and neuropsychological task performance and ratings of EF in 61 4-year-old preschool children (51 boys, 10 girls) with self-regulation difficulties were examined, with regression analyses controlling for the effects of ADHD inattention symptoms. In the study sample, higher teacher-rated SCT symptoms are significantly associated with poorer performance on tasks of visual-perceptual abilities, auditory and visual attention, sustained and selective attention, inhibitory control, pre-numerical/numerical concepts, and slower processing speed, but SCT symptoms are not significantly associated with working memory, attention shifting or cognitive flexibility when controlling for ADHD inattention. Higher parent-rated SCT symptoms are significantly associated with visual-perceptual abilities. ADHD inattention symptoms are more strongly associated than SCT with daily life EF ratings; neither parent- nor teacher-rated SCT symptoms are significantly associated with daily life ratings of inhibition, working memory, or planning/organization after controlling for ADHD inattention. This study suggests that SCT symptoms contribute to EF deficits at least on neurocognitive tasks assessing visual-perceptual/spatial abilities, attention to detail and processing speed, as observed in this sample of young children at risk for ADHD, and may be an important intervention target.

Immunohistochemical expression of platelet growth factor and vascular endothelial growth factor in patients with melanoma with and without redness (Brenner sign).

OBJECTIVE: To assess whether an erythematous eruption in the vicinity of or distant from a melanoma lesion might be related to the vascular endothelial growth factor, the platelet-derived endothelial cell growth factor, or both. METHODS: Biopsy specimens from 13 patients with primary melanoma, 6 of whom had erythematous eruptions and 7 who did not, were studied by immunohistochemistry for the expression of vascular endothelial growth factor and platelet-derived endothelial cell growth factor. RESULTS: Vascular endothelial growth factor was positive in 3 of 6 patients (50%) with melanoma and redness (Brenner sign) and in 4 of 7 patients (57%) with melanoma without redness. Platelet-derived endothelial cell growth factor was positive in all 6 patients (100%) with melanoma and redness and in 4 of 7 patients (57%) with melanoma without redness. CONCLUSION: Platelet-derived endothelial cell growth factor may have a part in the pathogenesis of the redness observed in patients with melanoma, called Brenner sign, by affecting vasculature function.

A survey of sex differences in 249 pemphigus patients and possible explanations.

BACKGROUND: Pemphigus, an immunoblistering disorder, is reported with equal or near equal frequencies in men and women despite prominent female predominance in prevalence ratios of the vast majority of autoimmune diseases. OBJECTIVE: To assess a possible correlation between pemphigus and intake of sex hormones in a cohort of pemphigus patients. METHODS: A prospective online survey using a specially designed questionnaire was conducted among patients with diagnosed pemphigus in the United States during a 1-year period from September 2005 through September 2006. RESULTS: A total of 249 pemphigus cases were enrolled, 158 women (63%) and 91 men (36%). The female-to-male ratio was 1.7:1. Age at onset of the disease ranged from 16 to 85 years, mean 4.5.4+/-12.9 [corrected] years for both sexes (not statistically significant): 45.3 for women and 45.7 for men. At the time of pemphigus diagnosis, 12% (20 of 158) of the women and 4% (4 of 91) of the men reported using hormone replacement therapy. At the time of disease onset, 46% (20 of 43) of the postmenopausal women took hormone replacement therapy. LIMITATIONS: Possible questionnaire self-reporting biases. CONCLUSIONS: The finding of a female predominance among pemphigus patients is attributed to the immunopathogenesis of the disease that makes women more susceptible to this and other autoimmune disorders and to the strikingly high proportion of hormone replacement therapy users found among postmenopausal women.

Drug rash with eosinophilia and systemic symptoms syndrome: sex and the causative agent.

The study was conducted to determine whether there is a sex difference in the causative agent of drug rash with eosinophilia and systemic symptoms (DRESS) syndrome. A retrospective review of hospital records from 1999 to 2006 in the dermatology department of a large municipal medical center found 8 patients who met the criteria for DRESS syndrome: drug-induced generalized eruption, associated systemic involvement (lymph node or visceral), and presence of eosinophilia (eosinophil count > or =1500/microL and/or circulating atypical lymphocytes). There were 4 men and 4 women, aged 19 to 53 years, with a mean age of 38 years. A sex difference was found in 3 parameters: (1) age, younger age in women; (2) time interval between drug intake and rash, shorter in women; and (3) the culprit drug: carbamazepine in 2 men and 2 women, salazopyrin in 2 women, and phenytoin in 2 men. While the sample is small, there is an indication of sex differences in the DRESS syndrome.

Severe acne vulgaris and tobacco smoking in young men.

As the relationship between tobacco smoking and acne remains unclear, we examined the relationship between cigarette smoking and severe acne in a large cohort of young men. Trained nurses interviewed subjects upon discharge from compulsory military service, regarding family history, habits, and tobacco smoking habits. Data was correlated with severe acne status, as diagnosed and coded by board-certified dermatologists. In total, 27,083 male subjects participated in the study from 1983 to 2003, of which 237 (0.88%) had severe acne, 11,718 (43.27%) were active smokers, and 15,365 (56.73%) were nonsmokers at the time of interviews. Active smokers showed a significantly lower prevalence of severe acne (0.71%) than nonsmokers (1.01%) (P = 0.0078). An inverse dose-dependent relationship between severe acne prevalence and daily cigarette consumption became significant from 21 cigarettes a day (chi2 and trend test: P < 0.0001), odds ratio: 0.2 (95% CI: 0.06-0.63). The study did not aim to establish a temporal correlation, and passive smoking and acne treatments were not measured. Previous in vitro and clinical studies strongly support an association with nicotine. We suggest a trial with topical nicotine treatment for acne to further investigate this association.

Significance of autoimmunity to non-desmoglein targets in pemphigus.

The pathogenesis of pemphigus vulgaris (PV) is a highly controversial, "hot" topic that has received considerable enrichment in recent years by both clinical and basic researchers. On the one hand, the classical view of desmogleins (Dsg) as main targets of this autoimmune disease is supported by the characterization of pathogenic anti-Dsg3 antibodies from both patients and animal models. On the other hand, fundamental doubt has been raised towards this monopathogenic view by several independent factors: (1) pemphigus lesions can be induced in Dsg3-knockout (KO) mice; (2) pemphigus sera contain multiple autoantibodies against different adhesion molecules and also cholinergic receptors; (3) experimental inhibition of PV IgG induced acantholysis can be obtained by interference with different signaling cascades regulating both calcium homeostasis and apoptosis; and (4) cholinergic agonists exhibit anti-acantholytic activity both in vitro and in vivo. The field is open for controlled clinical trials and further basic research to unfold the true story of the pemphigus enigma and provide the basis for a better treatment of pemphigus patients.

Increased use of mental health services related to isotretinoin treatment: a 5-year analysis.

BACKGROUND: The association between exposure to Isotretinoin, the development of depression and suicide attempts is controversial. OBJECTIVE: To retrospectively assess pattern of utilization of mental health services in the Israeli Defense Forces (IDF) during a 5-year period for all subjects exposed to Isotretinoin in comparison to a control group consisting of army conscripts suffering from psoriasis. All subjects were young adults (18 to 21 years old) in compulsory military service. METHOD: Exposure to Isotretinoin mandates reporting and marking as a coded medical profile in the IDFs' computerized medical record of each conscript and soldier. Medical data, tracked by military medical profiles, were summarized from medical records of all subjects treated by Isotretinion during the years 1999-2003 and for the control group for the same period. Use of mental health services was the a-priori defined primary outcome measure. RESULTS: During the study period 1419 subjects were exposed to Isotretinoin and 1102 suffered from psoriasis. Utilization of mental health services was highest for the index group wherein 17.2% (245/1419) of subjects were evaluated or treated compared to 12.5% in the control group (psoriasis). The inter-group differences were statistically significant; Chi-square=15.9 (df=2), p=0.0003. CONCLUSIONS: We suggest that psychiatric evaluation be regularly undertaken prior to initiation of Isotretinion treatment in young adults at risk, as well as providing follow-up visits during and at completion of treatment.

Multiple common naevi in Israeli adolescents.

The most prominent known phenotypic risk factor for the development of melanoma is a large number of common melanocytic naevi. The aims of this study were to determine the prevalence of multiple common naevi in a group of 17-year-olds and to identify possible associated risk factors. The study subjects were approximately 40,000 female and 60,000 male 17-year-old Jewish Israelis presenting to army recruitment centres for compulsory military service during a 1-year period from January 2003 to December 2003. Data were gathered on whole-body naevi count, years of education, place of residence and place of birth of the recruit, his/her parents and paternal grandparents. The count of whole-body naevi was carried out by military physicians. The multiple naevi phenotype was defined by the presence of more than 50 naevi. The prevalence of the phenotype was highest amongst recruits of American origin and lowest amongst those of Asian and African origin (P < 0.0001). The prevalence differed in different population centres, the highest being in the coastal cities of Haifa and Tel Aviv. This pattern corresponded to the geographical distribution of melanoma in Israel. Amongst male conscripts, the phenotype was more common in the better educated group (> or =12 years) than in the poorer educated conscripts (< or = 11 years) (P = 0.051). When adjusted for the factors studied, a significant correlation was found between a high multiple common naevi count and American and European origin and residence in coastal cities, reflecting the complex interplay between endogenous and exogenous factors of melanoma.