Live-imaging of revertant and therapeutically restored dystrophin in the DmdEGFP-mdx mouse model for Duchenne muscular dystrophy.

BACKGROUND: Dmdmdx , harbouring the c.2983C>T nonsense mutation in Dmd exon 23, is a mouse model for Duchenne muscular dystrophy (DMD), frequently used to test therapies aimed at dystrophin restoration. Current translational research is methodologically hampered by the lack of a reporter mouse model, which would allow direct visualization of dystrophin expression as well as longitudinal in vivo studies. METHODS: We generated a DmdEGFP-mdx reporter allele carrying in cis the mdx-23 mutation and a C-terminal EGFP-tag. This mouse model allows direct visualization of spontaneously and therapeutically restored dystrophin-EGFP fusion protein either after natural fibre reversion, or for example, after splice modulation using tricyclo-DNA to skip Dmd exon 23, or after gene editing using AAV-encoded CRISPR/Cas9 for Dmd exon 23 excision. RESULTS: Intravital microscopy in anaesthetized mice allowed live-imaging of sarcolemmal dystrophin-EGFP fusion protein of revertant fibres as well as following therapeutic restoration. Dystrophin-EGFP-fluorescence persisted ex vivo, allowing live-imaging of revertant and therapeutically restored dystrophin in isolated fibres ex vivo. Expression of the shorter dystrophin-EGFP isoforms Dp71 in the brain, Dp260 in the retina, and Dp116 in the peripheral nerve remained unabated by the mdx-23 mutation. CONCLUSION: Intravital imaging of DmdEGFP-mdx muscle permits novel experimental approaches such as the study of revertant and therapeutically restored dystrophin in vivo and ex vivo.

Skin diseases are more common than we think: screening results of an unreferred population at the Munich Oktoberfest.

BACKGROUND: Skin diseases are ranked as the fourth most common cause of human illness, resulting in an enormous non-fatal burden. Despite this, many affected people do not consult a physician. Accordingly, the actual skin disease burden might be even higher since reported prevalence rates are typically based on secondary data that exclude individuals who do not seek medical care. OBJECTIVE: The aim of the study was to investigate the prevalence of skin diseases in an unreferred population in a real-life setting. METHODS: A cross-sectional study of 9 days duration was performed in 2016 at the 'Bavarian Central Agricultural Festival', which is part of the Munich Oktoberfest. As part of a public health check-up, screening examinations were performed randomly on participating visitors. All participants were 18 years or older and provided written informed consent. RESULTS: A total of 2701 individuals (53.5% women, 46.2% men; mean age 51.9 ± 15.3 years) participated in the study. At least one skin abnormality was observed in 1662 of the participants (64.5%). The most common diagnoses were actinic keratosis (26.6%), rosacea (25.5%) and eczema (11.7%). Skin diseases increased with age and were more frequent in men (72.3%) than in women (58.0%). Clinical examinations showed that nearly two-thirds of the affected participants were unaware of their abnormal skin findings. CONCLUSION: Skin diseases might be more common than previously estimated based on the secondary data of some sub-populations. Further information and awareness campaigns are needed to improve people's knowledge and reduce the global burden associated with skin diseases.

[Health care of chronic inflammatory skin diseases : Do affected individuals seek dermatological care?] / Versorgung von chronisch entzündlichen Hauterkrankungen : Gehen Betroffene zum niedergelassenen Dermatologen?

BACKGROUND: Psoriasis, atopic eczema and urticaria are chronic inflammatory skin diseases that are often associated with an impairment of affected individuals and their families. Despite constant progress in therapy of these diseases, affected people often do not consult an office-based dermatologist. OBJECTIVES: The aim of this study was to estimate which proportion of affected individuals with severe forms of these diseases receive treatment by an office-based dermatologist in Bavaria. MATERIALS AND METHODS: All dermatologists listed in the database of the Bavarian Association of Panel Doctors (KVB; Kassenärztliche Vereinigung Bayern; n = 499) were invited to participate in a paper-based cross-sectional study. The stated number of patients by each dermatologist were set in relation with the literature-based 1­year prevalence, as well as data on population and data of the KVB. Estimations were based on three approaches (conservative, medium, and progressive estimation method). RESULTS: Overall, 137 dermatologists participated (38.7% women; mean age: 53.2 ± 8.5 years). Conservative estimation indicated that 56.5% of individuals with moderate to severe psoriasis, 57.3% of individuals with moderate to severe atopic eczema and 71.9% of those suffering from chronic spontaneous urticaria are not seen by an office-based dermatologist. CONCLUSION: Many affected individuals seem not to seek an office-based dermatologist when affected by a severe skin condition. Thus, further and more precise studies to identify, address and minimize barriers to optimal patient care are needed.

Newly acquired kiwi fruit allergy after bone marrow transplantation from a kiwi-allergic donor.

BACKGROUND: The phenomenon of allergy transfer from an allergic donor to a non-allergic recipient via hematopoietic cell transplantation has been described by several reports. However, it could not yet been conclusively shown that allergic reaction of the recipient is elicited by the donor's cells. OBJECTIVES: In the case of a 46-year-old male patient who - for the first time in his life - had two episodes of oral allergic syndrome upon kiwi consumption after having received myeloablative hematopoietic stem cell transplantation (HCT) from his kiwi-allergic sister, we aimed to clarify the origin of allergen reactive cells in the donor. We not only intended to demonstrate if allergy was transferred by HCT but also to present an experimental workup for the analysis of allergy transfer by HCT. METHODS: Allergic sensitization to kiwi in recipient and donor was proven by ImmunoCAP. Furthermore, origin of peripheral blood mononuclear cells (PBMCs) was analyzed by chromosomal fluorescence in situ hybridization (FISH). To confirm allergic reaction and activation of hematopoietic cells by customized kiwi extract, we performed basophil activation test from whole blood as well as T cell proliferation assays from purified PBMCs of both recipient and donor. RESULTS: Basophil activation upon kiwi extract was demonstrated in both recipient and donor. Besides, we showed proliferation of CD4(+) T cells after incubation with kiwi extract. FISH analysis proved that hematopoietic cells of the male recipient completely originated from the female donor. CONCLUSION: Exemplified in this patient, we show for the first time that allergy transfer is mediated by the donor's cells. Moreover, our experimental approach using customized kiwi extract to prove contribution of kiwi-specific T and B cells in both kiwi-allergic recipient and donor could serve as a model approach for future studies.

Bridging classical and quantum mechanics.

Using a watt balance and a frequency comb, a mass-energy equivalence is derived. The watt balance compares mechanical power measured in terms of the meter, the second, and the kilogram to electrical power measured in terms of the volt and the ohm. A direct link between mechanical action and the Planck constant is established by the practical realization of the electrical units derived from the Josephson and the quantum Hall effects. By using frequency combs to measure velocities and acceleration of gravity, the unit of mass can be realized from a set of three defining constants: the Planck constant h, the speed of light c, and the hyperfine splitting frequency of 133Cs.

[Cerebrovascular complications of immunologically mediated heparin-induced thrombocytopenia]. / Zerebrovaskuläre Komplikationen der Heparininduzierten Thrombozytopenie Typ II (HIT-II).

Immunologically mediated heparin-induced thrombocytopenia (HIT) is a thrombotic disease caused by antibodies occurring after heparin exposure. Thrombocytopenia occurs within a few days after heparin exposure, about half of HIT-patients develop venous or arterial thrombotic complications. Neurological complications of HIT are mainly ischaemic stroke and sinus vein thrombosis. To ensure the primary clinical diagnosis functional and immunological assays for antibody detection are available. The probability for the occurrence of HIT depends on the nature of heparin employed (LMWH vs. UFH) and individual patient characteristics such as gender and primary disease (medical vs. surgical patients). In the case of suspected HIT heparin administration should be discontinued immediately and replaced by an alternative anticoagulation to prevent the expansion or development of further thrombotic complications. Herein we report a case of a patient suffering from HIT-associated embolic cerebral ischaemic stroke.

Constraints on spin-dependent short-range interaction between nucleons.

We search for a spin-dependent P- and T-violating nucleon-nucleon interaction mediated by light pseudoscalar bosons such as axions or axionlike particles. We employ an ultrasensitive low-field magnetometer based on the detection of free precession of colocated 3He and 129Xe nuclear spins using SQUIDs as low-noise magnetic flux detectors. The precession frequency shift in the presence of an unpolarized mass was measured to determine the coupling of pseudoscalar particles to the spin of the bound neutron. For boson masses between 2 and 500 µeV (force ranges between 3×1(-4) m and 10(-1) m) we improved the laboratory upper bounds by up to 4 orders of magnitude.

Stabilized high-power laser system for the gravitational wave detector advanced LIGO.

An ultra-stable, high-power cw Nd:YAG laser system, developed for the ground-based gravitational wave detector Advanced LIGO (Laser Interferometer Gravitational-Wave Observatory), was comprehensively characterized. Laser power, frequency, beam pointing and beam quality were simultaneously stabilized using different active and passive schemes. The output beam, the performance of the stabilization, and the cross-coupling between different stabilization feedback control loops were characterized and found to fulfill most design requirements. The employed stabilization schemes and the achieved performance are of relevance to many high-precision optical experiments.

Current CONtrolled Transmit And Receive Coil Elements (CONTAR) for Parallel Acquisition and Parallel Excitation Techniques at High-Field MRI.

A novel intrinsically decoupled transmit and receive radio-frequency coil element is presented for applications in parallel imaging and parallel excitation techniques in high-field magnetic resonance imaging. Decoupling is achieved by a twofold strategy: during transmission elements are driven by current sources, while during signal reception resonant elements are switched to a high input impedance preamplifier. To avoid B(0) distortions by magnetic impurities or DC currents a resonant transmission line is used to relocate electronic components from the vicinity of the imaged object. The performance of a four-element array for 3 T magnetic resonance tomograph is analyzed by means of simulation, measurements of electromagnetic fields and bench experiments. The feasibility of parallel acquisition and parallel excitation is demonstrated and compared to that of a conventional power source-driven array of equivalent geometry. Due to their intrinsic decoupling the current-controlled elements are ideal basic building blocks for multi-element transmit and receive arrays of flexible geometry.

Glutamate as a spectroscopic marker of hippocampal structural plasticity is elevated in long-term euthymic bipolar patients on chronic lithium therapy and correlates inversely with diurnal cortisol.

While an excess of glucocorticoids is associated with hippocampal pathology in mood disorders, lithium exerts robust neuroprotective and neurotrophic effects. Here, 21 stably remitted bipolar I patients who had been on chronic lithium maintenance therapy, on average, for more than a decade, and 19 carefully matched healthy controls were studied using 3 T (1)H-magnetic resonance spectroscopy of left and right hippocampus. Salivary cortisol samples were obtained to assess activity of the hypothalamus-pituitary-adrenal system. Absolute concentrations of N-acetylaspartate (NAA), choline-containing compounds and total creatine were similar in euthymic bipolar patients and healthy controls. Hippocampal glutamate concentrations were significantly increased as an effect of patient status (patients>controls) and laterality (left hippocampus>right hippocampus). Hippocampal glutamate content (Glu) was strongly correlated with NAA. Across groups and within the patient group, diurnal saliva cortisol levels showed a significant inverse relationship with both Glu and NAA. Taken together, these results add to the concept of bipolar disorder as an illness involving disturbed hippocampal structural plasticity under the opposing influences of lithium and glucocorticoids.

Complex regional pain syndromes: new pathophysiological concepts and therapies.

Complex regional pain syndrome (CRPS), formerly known as Sudeck's dystrophy and causalgia, is a disabling and distressing pain syndrome. We here provide a review based on the current literature concerning the epidemiology, etiology, pathophysiology, diagnosis, and therapy of CRPS. CRPS may develop following fractures, limb trauma or lesions of the peripheral or CNS. The clinical picture comprises a characteristic clinical triad of symptoms including autonomic (disturbances of skin temperature, color, presence of sweating abnormalities), sensory (pain and hyperalgesia), and motor (paresis, tremor, dystonia) disturbances. Diagnosis is mainly based on clinical signs. Several pathophysiological concepts have been proposed to explain the complex symptoms of CRPS: (i) facilitated neurogenic inflammation; (ii) pathological sympatho-afferent coupling; and (iii) neuroplastic changes within the CNS. Furthermore, there is accumulating evidence that genetic factors may predispose for CRPS. Therapy is based on a multidisciplinary approach. Non-pharmacological approaches include physiotherapy and occupational therapy. Pharmacotherapy is based on individual symptoms and includes steroids, free radical scavengers, treatment of neuropathic pain, and finally agents interfering with bone metabolism (calcitonin, biphosphonates). Invasive therapeutic concepts include implantation of spinal cord stimulators. This review covers new aspects of pathophysiology and therapy of CRPS.

Central mechanisms of experimental and chronic neuropathic pain: findings from functional imaging studies.

Over the last few years remarkable efforts have been made using functional imaging studies to unravel brain processing of pain and decipher underlying neuronal mechanisms. Cerebral processing in experimental pain models, especially those provoking hyperalgesia, and its pharmacological modulation will form the first part of this review. In a second part we will address central mechanisms of clinical neuropathic pain. Up to now, there are at least six main mechanisms involved in the chronification of neuropathic pain: (i) activity increase in areas of the pain neuromatrix, (ii) recruitment of additional cortical areas beyond the classical pain neuromatrix, (iii) cortical reorganization and maladaptive neuroplasticity, (iv) alterations in neurochemistry (v) structural brain changes and (vi) disruption of the brain default mode network. In a third part of this review we discuss mechanisms of endogenous pain modulation.

[Neuropathic pain and neuroplasticity in functional imaging studies]. / Neuropathische Schmerzsyndrome und Neuroplastizität in der funktionellen Bildgebung.

Neuropathic pain syndromes are characterised by the occurrence of spontaneous ongoing and stimulus-induced pain. Stimulus-induced pain (hyperalgesia and allodynia) may result from sensitisation processes in the peripheral (primary hyperalgesia) or central (secondary hyperalgesia) nervous system. The underlying pathophysiological mechanisms at the nociceptor itself and at spinal synapses have become better understood. However, the cerebral processing of hyperalgesia and allodynia is still controversially discussed. In recent years, neuroimaging methods (functional magnetic resonance imaging, fMRI; magnetoencephalography, MEG; positron emission tomography, PET) have provided new insights into the aberrant cerebral processing of neuropathic pain. The present paper reviews different cerebral mechanisms contributing to chronicity processes in neuropathic pain syndromes. These mechanisms include reorganisation of cortical somatotopic maps in sensory or motor areas (highly relevant for phantom limb pain and CRPS), increased activity in primary nociceptive areas, recruitment of new cortical areas usually not activated by nociceptive stimuli and aberrant activity in brain areas normally involved in descending inhibitory pain networks. Moreover, there is evidence from PET studies for changes of excitatory and inhibitory transmitter systems. Finally, advanced methods of structural brain imaging (voxel-based morphometry, VBM) show significant structural changes suggesting that chronic pain syndromes may be associated with neurodegeneration.

Kinetics of the fe2+-mg, order-disorder reaction in anthophyllites: quantitative cooling rates.

The kinetics of the Fe(2+)-Mg, order-disorder phenomenon in a highly ordered natural anthophyllite have been determined over the temperature range from 400 degrees to 720 degrees C at a pressure of 2 kilobars. At temperatures of 600 degrees C and above, equilibrium is attained by disordering as well as ordering reactions. The intracrystalline exchange is defined by a standard Gibbs free energy of 4247 +/- 54 calories per formula unit. Rate studies at 550 degrees and 500 degrees C show that equilibrium is attained by ordering but not by disordering within the same time scale and that the exchange reaction is characterized by an activation energy of approximately 55 kilocalories per formula unit. An equilibration temperature for the natural anthophyllite of 270 degrees C is determined from the termination of the ordering process owing to excessively slow reaction kinetics after approximately 10(7) years. From the rate constants of the exchange process, for different crystallization temperatures, the apparent equilibration temperature of 270 degrees C defines a maximum linear cooling rate for the rock of 1 x 10(4) degrees C per year.

In vivo detection of hepatitis C virus (HCV) RNA in the brain in a case of encephalitis: evidence for HCV neuroinvasion.

We report here a 27-year-old woman who presented with encephalitis of unknown origin. Magnetic resonance imaging (MRI) of the brain revealed leukoencephalopathy, cerebrospinal fluid showed signs of inflammation. Serum and brain biopsy tissue was tested positive for hepatitis C virus (HCV). Neuropathological investigation supported the hypothesis of viral encephalitis. C3, C4 and cryoglobulins as well as cerebral MR-angiography were normal. Neurological complications of HCV infection other than hepatic encephalopathy are generally attributed to parainfectious phenomena. This is the first case of HCV-RNA detection in vivo in human brain in literature and it raises the possibility that HCV is able to induce encephalitis caused by neurotrophism. This is supported by the fact that there is a growing body of literature on HCV-induced cerebral dysfunction and laboratory findings indicating HCV neuroinvasion.

Acute onset of fatal vegetative symptoms: unusual presentation of adult Alexander disease.

Since genetic analysis of the GFAP gene for the diagnosis of adult Alexander disease (AD) has been established in 2001, several cases of both sporadic and familial cases of AD have been described. Except for one patient, all subjects revealed glial fibrillary acidic protein (GFAP) mutations, and clinical progression of symptoms, mainly bulbar and pseudobulbar, were moderate. Here we report on a patient with acute onset of vegetative symptoms, rapid progression, and death within 2 months. Although histology and final magnetic resonance imaging (MRI) were characteristic of AD, sequencing of the encoding GFAP gene revealed no mutation. We believe that this case report expands the so far known clinical spectrum and MRI dynamics of adult AD, and suggest that analysis of the coding part of GFAP may be inconclusive in rare cases. In such patients, only histology may lead to definitive diagnosis.

Microcystic adnexal carcinoma (MAC)-like squamous cell carcinoma as a differential diagnosis to Bell´s palsy: review of guidelines for refractory facial nerve palsy.

BACKGROUND: Bell´s palsy is the most common cause of facial paralysis worldwide and the most common disorder of the cranial nerves. It is a diagnosis of exclusion, accounting for 60-75% of all acquired peripheral facial nerve palsies. Our case shows the first case of a microcystic adnexal carcinoma-like squamous cell carcinoma as a cause of facial nerve palsy. CASE PRESENTATION: The patient, a 70-year-old Caucasian male, experienced subsequent functional impairment of the trigeminal and the glossopharyngeal nerve about 1½ years after refractory facial nerve palsy. An extensive clinical work-up and tissue biopsy of the surrounding parotid gland tissue was not able to determine the cause of the paralysis. Primary infiltration of the facial nerve with subsequent spreading to the trigeminal and glossopharyngeal nerve via neuroanastomoses was suspected. After discussing options with the patient, the main stem of the facial nerve was resected to ascertain the diagnosis of MAC-like squamous cell carcinoma, and radiochemotherapy was subsequently started. CONCLUSION: This case report shows that even rare neoplastic etiologies should be considered as a cause of refractory facial nerve palsy and that it is necessary to perform an extended diagnostic work-up to ascertain the diagnosis. This includes high-resolution MRI imaging and, as perilesional parotid biopsies might be inadequate for rare cases like ours, consideration of a direct nerve biopsy to establish the right diagnosis.

Hippocampal glutamate concentration predicts cerebral theta oscillations during cognitive processing.

RATIONALE: Brain waves reflect collective behavior of neurons and provide insight into distributed network processing. Frontal and hippocampal theta oscillations (4-7 Hz) were linked to cognitive tasks and animal studies have suggested an involvement of glutamatergic neurotransmission in integrative frontal-hippocampal processing. Human evidence for such relationships is lacking. METHODS: Here, we studied the associations between glutamate concentrations in the hippocampal region, measured by a 3-T proton magnetic resonance spectroscopy (1H-MRS), and EEG theta activity during an auditory target detection paradigm. RESULTS: A robust relationship between hippocampal glutamate and frontal theta activity during stimulus processing was found. Moreover, frontal theta oscillations were related to response speed. CONCLUSION: The results suggest a functional coupling between the frontal cortex and hippocampal region during stimulus processing and support the idea of the hippocampus as a neural rhythm generator driven by glutamatergic neurotransmission. These preliminary data show, for the first time, a relationship between in vivo measured glutamate and basic cerebral information processing in humans.