Centrosome amplification is a frequent phenomenon in malignancies and may facilitate tumorigenesis by promoting chromosomal instability. On the other hand, a centrosome inactivation checkpoint ...comprising centrosome amplification leading to elimination of cells by mitotic catastrophe has been described in response to DNA damage by ionizing radiation or cytostatic drugs. So far, the exact nature of DNA damage-induced centrosome amplification, which might be overduplication or fragmentation of existing centrosomes, has been controversial. To solve this controversy, we have established a method to distinguish between these two possibilities using A549 cells expressing photoconvertible CETN2-Dendra2. In response to various DNA-damaging treatments, centrosome amplification but not fragmentation was observed. Moreover, centrosome amplification was preceded by excessive formation of centrin-containing centriolar satellites, which were identified as de novo-generated atypical centrin dots staining positive for centriolar satellite markers but negative or only weakly positive for other established centrosomal markers, and which could be verified as centriolar satellites using immunogold electron microscopy. In line with this notion, disruption of dynein-mediated recruitment of centrosomal proteins via centriolar satellites suppressed centrosome amplification after DNA damage, and excessive formation of centriolar satellites could be inhibited by interference with Chk1, a known mediator of centrosome amplification in response to DNA damage. In conclusion, we provide a model in which a Chk1-mediated DNA damage checkpoint induces excessive formation of centriolar satellites constituting assembly platforms for centrosomal proteins, which subsequently leads to centrosome amplification.
Introduction
Developmental predisposition to schizophrenia can be a consequence of early experienced traumas. Transgenerational trauma is process in which traumatic experience of one generation is ...passed on to the next generation.
Objectives
To show connection between transgenerational transmission of trauma and development of schizophrenia.
Methods
Psychiatric interview, psychological testing.
Results
Patient G.E. age 29, admitted to Psychiatry Clinic due to altered behavior, aggressiveness and presence of delusions and hallucinations. First mental problems in form of a catatonic stupor appeared 6 years ago. Patient has history of earlier abuse of psychoactive substances. A drug test performed at admission was negative. Patient was born in Srebrenica in 1993, he escaped to Tuzla with his mother in July 1995, while father survived escaping on foot. Patient is a first child from his father’s second marriage. The father’s first wife and two minor children were shot by Bosnian Serbs in early 1992. Patient was born a year and a half after death of his siblings and was named after his half-sister. Patient’s father consumed alcohol after the war and was aggressive towards children. In the last two years, patient had frequent hallucinations, he told his parents that voices were telling him to kill his mother and told his father that his children were still alive. Diagnostic processing was performed and diagnosis of schizophrenia was stated. During hospitalization, patient was treated with olanzapine and low doses of haloperidol, along with haloperidol decanoate, which resulted in significant reduction of productive psychotic symptoms. A partial remission is achieved, negative schizophrenic symptoms and cognitive impairments verified by psychological instruments remain.
Conclusions
Case report emphasize transgenerational transmission of trauma: father‘s untreated trauma, alcohol dependency and abuse of the patient in childhood. These findings are important for treatment and therapeutic considerations. Mentalizing is a possible mediator between childhood abuse and negative symptoms. Parental bonding was explored within high expressed emotions theory as a risk factor for relapse to psychosis, especially the “affectionless control” in the parental (mainly father‘s) bonding style. Studies also stated that psychotic patients often show insecure attachment representations. Possible pathway for further analysis could be discussed: a cold parental bonding style leading to experienced emotional neglect and attachment avoidance might be reflected in lower capacity to mentalize. To improve the mentalization capacity, it would be essential to establish a sustainable therapeutic treatment frame.
Disclosure of Interest
None Declared
Mutations in
SCN2A
cause epilepsy syndromes of variable severity including neonatal-infantile seizures. In one case, we previously described additional childhood-onset episodic ataxia. Here, we ...corroborate and detail the latter phenotype in three further cases. We describe the clinical characteristics, identify the causative
SCN2A
mutations and determine their functional consequences using whole-cell patch-clamping in mammalian cells. In total, four probands presented with neonatal-onset seizures remitting after five to 13 months. In early childhood, they started to experience repeated episodes of ataxia, accompanied in part by headache or back pain lasting minutes to several hours. In two of the new cases, we detected the novel mutation p.Arg1882Gly. While this mutation occurred de novo in both patients, one of them carries an additional known variant on the same
SCN2A
allele, inherited from the unaffected father (p.Gly1522Ala). Whereas p.Arg1882Gly alone shifted the activation curve by −4 mV, the combination of both variants did not affect activation, but caused a depolarizing shift of voltage-dependent inactivation, and a significant increase in Na
+
current density and protein production. p.Gly1522Ala alone did not change channel gating. The third new proband carries the same de novo
SCN2A
gain-of-function mutation as our first published case (p.Ala263Val). Our findings broaden the clinical spectrum observed with
SCN2A
gain-of-function mutations, showing that fairly different biophysical mechanisms can cause a convergent clinical phenotype of neonatal seizures and later onset episodic ataxia.
Summary
Background Tape stripping is a common method for investigating stratum corneum (SC) physiology as well as bioavailability and bioequivalence of topical drugs.
Objectives To investigate the ...influence of procedures (anatomical site, pressure, pressure duration, tape removal rate) inherent in each stripping protocol on changes in skin physiology.
Methods Tape stripping was performed using tapes on the forearm, forehead and back. On the forearm different pressures (165 and 330 g cm−2), durations of pressure (2 and 10 s), and removal rate (slow and rapid removal) were used. Changes in skin physiology were evaluated by measurement of transepidermal water loss (TEWL) and hydration.
Results A significant influence of all parameters on the TEWL increase as a function of tape strip number was observed. The fastest increase was demonstrated on the forehead, followed by the back and, lastly, the forearm. Rapid removal produced a protracted increase in comparison with slow removal. Pressure for 10 s induced a faster increase in TEWL than 2 s pressure. Likewise, pressure at 330 g cm−2 induced an earlier increase than pressure at 165 g cm−2. Skin hydration was not influenced by the variables tested.
Conclusions Tape stripping results are influenced dramatically by all investigated parameters. A dynamic SC stress test to investigate SC cohesion more closely is proposed based on the present observations.
Here we describe some of the crucial steps to generate induced pluripotent stem cells (iPSCs) using mRNA transfection. Our approach uses a V. virus-derived capping enzyme instead of a cap-analog, ...ensuring 100% proper cap orientation for in vitro transcribed mRNA. V. virus' 2′-O-Methyltransferase enzyme creates a cap1 structure found in higher eukaryotes and has higher translation efficiency compared to other methods. Use of the polymeric transfection reagent polyethylenimine proved superior to other transfection methods.
The mRNA created via this method did not trigger an intracellular immune response via human IFN-gamma (hIFN-γ) or alpha (hIFN-α) release, thus circumventing the use of suppressors. Resulting mRNA and protein were expressed at high levels for over 48h, thus obviating daily transfections. Using this method, we demonstrated swift activation of pluripotency associated genes in human fibroblasts. Low oxygen conditions further facilitated colony formation. Differentiation into different germ layers was confirmed via teratoma assay.
Reprogramming with non-synthetic mRNA holds great promise for safe generation of iPSCs of human origin. Using the protocols described herein we hope to make this method more accessible to other groups as a fast, inexpensive, and non-viral reprogramming approach.
Balanced chromosomal rearrangements define distinct entities in acute myeloid leukemia (AML). Here, we present 13 AML cases with t(8;16)(p11;p13) with observed low incidence (13/6124 patients), but ...more frequent presentation in therapy-related AML than in de novo AML (7/438 versus 6/5686, P=0.00001). Prognosis was poor with median overall survival of 4.7 months. Cytomorphology was characterized by parallel positive myeloperoxidase and non-specific esterase staining, therefore, French-American-British (FAB)-classification was impossible and origin of the AML with t(8;16) from an early stem cell with myeloid and monoblastic potential is hypothesized. Erythrophagocytosis was observed in 7/13 cases. Using gene expression profiling on 407 cases, patients with t(8;16) were compared to AML FAB subtypes with normal karyotype. Principal component analyses demonstrated that AML with t(8;16) were distinct from FAB subtypes M1, M4, M5a/b. When further compared to AML showing balanced rearrangements, that is, current WHO categories t(15;17), t(8;21), inv(16) and t(11q23)/MLL, AML with t(8;16) cases were clustered close to t(11q23)/MLL sharing commonly expressed genes. Subsequently, a pairwise comparison discriminated AML with t(8;16) from AML with t(11q23)/MLL, thus defining a highly unique signature for AML with t(8;16). In conclusion, AML with t(8;16) demonstrates unique cytomorphological, cytogenetic, molecular and prognostic features and is a specific subtype of AML.
Summary
Background
As previous observations have indicated an inter‐relationship between irritant and allergic skin reactions we analysed data of synchronous allergen and sodium lauryl sulfate (SLS) ...patch tests in terms of a relationship between SLS responsiveness and allergic patch test reactions.
Objectives
To analyse differences in terms of allergen‐specific and overall reaction profiles between patients with vs. those without an irritant reaction to SLS.
Methods
Clinical data of 26 879 patients patch tested from 2008 to 2011 by members of the Information Network of Departments of Dermatology were analysed. After descriptive analyses, including the MOAHLFA index, the positivity ratio and the reaction index, a negative binomial hurdle model was adopted to investigate the correlation between SLS reactivity and positive patch test reactions.
Results
Men, patients aged ≥ 40 years and patients with an occupational dermatitis background were over‐represented in the SLS‐reactive group. Patients with an irritant reaction to SLS showed a higher proportion of weak positive reactions, as well as more questionable and irritant reactions to contact allergens than patients not reactive to SLS. The risk of an additional positive patch test reaction increased by 22% for SLS‐reactive patients compared with those who were SLS negative.
Conclusions
The marked association between SLS reactivity and the number of positive reactions in patch test patients may be due to nonspecific increased skin reactivity at the moment of patch testing only. However, increased SLS reactivity could also be due to longer‐lasting enhanced skin irritability, which may have promoted (poly‐)sensitization. Further studies, for example with longitudinal data on patients repeatedly patch tested with SLS and contact allergens, are necessary.
What's already known about this topic?
Irritation and contact allergy follow a partly similar pathomechanism and lead to similar morphology, e.g. in patch testing.
Doubtful and irritant, but also partly weak positive, patch test reactions to allergens have been found to be associated with irritant reactions to sodium lauryl sulfate (SLS) 0·25% in water.
Some studies have linked skin irritability as diagnosed with SLS to susceptibility to contact sensitization.
What does this study add?
Skin irritability to SLS is associated with multiple positive reactions to patch tests of the baseline series. The risk of having one additional positive reaction increases by 22% in patients with an irritant reaction to SLS.
It is apparent that further research beyond the mere analysis of patch test data is needed to disentangle the conflicting interpretations of irritant SLS reactions being indicative of false (weak) positive patch test reactions vs. increased susceptibility to contact sensitization.
Summary
Background There is evidence that a higher skin susceptibility may induce nonspecific erythematous or weak positive reactions to contact allergens in patch testing.
Objectives To evaluate ...whether simultaneous application of sodium lauryl sulphate (SLS) along with diagnostic patch tests with contact allergens can provide information regarding skin irritability which may help to discriminate allergic from nonspecific irritant reactions to contact allergens.
Methods Between July 2001 and June 2003, this prospective study collected patch test data of 5971 patients from 19 centres in Germany and Austria in the Information Network of Departments of Dermatology (IVDK). In addition to contact allergens (standard series and eight known ‘problematic’ allergens with a low reaction index and a high positivity ratio: 1,3‐diphenylguanidine, amerchol L‐101, benzalkonium chloride, benzoyl peroxide, cocamidopropyl betaine, octyl gallate, phenyl mercuric acetate and propylene glycol), patches with SLS 0·5% and 0·25% aq. were applied. Reactions to the allergens and to SLS were analysed at the IVDK data centre. The association between an erythematous or positive reaction to a certain allergen and an irritant reaction to SLS was assessed with logistic regression analysis, at the same time controlling for the influence of age and sex.
Results Of the 29 allergens of the standard series, 23 and 21 gave a higher percentage of nonspecific erythematous reactions in patients with an irritant reaction to 0·25% and 0·5% SLS, respectively, in comparison with SLS‐negative patients. All eight ‘problematic’ allergens gave an increased percentage of nonspecific erythematous reactions. Similarly, 22 and 21 allergens of the standard series gave a higher percentage of positive allergic reactions in patients with an irritant reaction to 0·25% and 0·5% SLS, respectively, and seven of the eight ‘problematic’ allergens gave a higher percentage of positive allergic rections (exception: octyl gallate). For most allergens, the markers of skin reaction (reaction index and positivity ratio) were worse in SLS‐positive patients. Differences were more pronounced when testing with SLS 0·25% than with SLS 0·5%.
Conclusions Because there is a convincing association between skin irritability (evaluated by SLS test) and the degree of skin reaction to contact allergens, the SLS test may help in deciding whether a doubtful erythematous or weakly ‘positive’ skin reaction should be interpreted as allergic or irritant.
The cutaneous reaction to detergents follows distinct kinetic rules: the duration of application and the irritant concentration are of major importance. The aim of this study was to evaluate the ...differences in kinetics of skin reaction between the standard irritant sodium lauryl sulfate (SLS), and 2 modern detergents: sodium laureth sulfate (SLES) and alkyl polyglucoside (APG). We performed patch testing with SLS and SLES (or APG) at different concentrations (0.125, 0.25, 0.5, 1.0 and 2.0%) and with different exposure times (6, 12 and 24 h). Evaluation was conducted by measurement of transepidermal water loss (TEWL) and laser Doppler flowmetry (LD) 24 h, 7 and 10 days after patch removal. We found a pronounced reaction to SLS, and a far milder one to SLES. Even at the highest concentration the skin reaction to APG was hard to detect. During the regeneration period (day 3–10) SLS showed even at day 10 an increased TEWL at all concentrations tested. The irritation due to SLES was convincingly detectable only up to day 7, whereas the APG‐tested skin areas showed no significant reaction even at day 3. These results demonstrate the improvement in reduction of skin irritation achieved by development of novel detergents.
Background
Aluminium tubes for pharmaceutical use are internally lacquered with epoxy resins (ER) based on bisphenol A diglycidyl ether (BADGE). Recently, it was shown that remnants of ER ...polymerization like BADGE are extractable from epoxy‐based coatings of commercially available tubes and may leach into semi‐solid drug preparations. We aimed to evaluate the safety of BADGE‐contaminated macrogol ointments in individuals sensitized to ER based on BADGE by use tests.
Methods
Repeated open application testing (ROAT) in 11 patients sensitized to ER based on BADGE with BADGE in macrogol ointments (3 mg/kg; 30 mg/kg, equivalent to BADGE concentration determined in macrogol ointment after storage in a commercially available tube; 300 mg/kg).
Results
The 30 mg/kg BADGE ointment elicited reactions in three patients, and another three patients reacted to 300 mg/kg BADGE ointment. No reactions to the vehicle control and 3 mg/kg BADGE were observed.
Conclusions
Elevated BADGE concentrations in ER‐coated aluminium tubes pose a risk of developing contact dermatitis to patients sensitized to ER based on BADGE. Quality standards are deemed necessary for the production of ER‐coated aluminium tubes intended for pharmaceutical use and should consider the results of the present ROAT study.
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