Revisiting adult neurogenesis and the role of erythropoietin for neuronal and oligodendroglial differentiation in the hippocampus.

Recombinant human erythropoietin (EPO) improves cognitive performance in neuropsychiatric diseases ranging from schizophrenia and multiple sclerosis to major depression and bipolar disease. This consistent EPO effect on cognition is independent of its role in hematopoiesis. The cellular mechanisms of action in brain, however, have remained unclear. Here we studied healthy young mice and observed that 3-week EPO administration was associated with an increased number of pyramidal neurons and oligodendrocytes in the hippocampus of ~20%. Under constant cognitive challenge, neuron numbers remained elevated until >6 months of age. Surprisingly, this increase occurred in absence of altered cell proliferation or apoptosis. After feeding a 15N-leucine diet, we used nanoscopic secondary ion mass spectrometry, and found that in EPO-treated mice, an equivalent number of neurons was defined by elevated 15N-leucine incorporation. In EPO-treated NG2-Cre-ERT2 mice, we confirmed enhanced differentiation of preexisting oligodendrocyte precursors in the absence of elevated DNA synthesis. A corresponding analysis of the neuronal lineage awaits the identification of suitable neuronal markers. In cultured neurospheres, EPO reduced Sox9 and stimulated miR124, associated with advanced neuronal differentiation. We are discussing a resulting working model in which EPO drives the differentiation of non-dividing precursors in both (NG2+) oligodendroglial and neuronal lineages. As endogenous EPO expression is induced by brain injury, such a mechanism of adult neurogenesis may be relevant for central nervous system regeneration.

[Beyond antibiotic therapy - Future antiinfective strategies - Update 2017]. / "Beyond antibiotic therapy" ­ Zukünftige antiinfektiöse Strategien ­ Update 2017.

BACKGROUND: The key elements in the therapy of surgical site infections (SSI) are surgical debridement and local and systemic antibiotic therapy; however, due to increasing antibiotic resistance, the development of additional therapeutic measures is of great interest for future trauma and orthopedic surgery. METHOD: Against the background of our own experimental and clinical experiences and on the basis of the current literature, possible future anti-infective strategies were elaborated. RESULTS/CONCLUSIONS: Bacteriophages were discovered and clinically implemented approximately one century ago and have been used in Western Europe for about one decade. They are currently used mainly in patients with burn injuries. It is likely that bacteriophages will become of great importance in view of the increasing antibiotic multi-drug resistance; however, they will probably not entirely replace antibiotic drugs. A combined use of bacteriophages and antibiotics is likely to be a more reasonable efficient therapy. In addition, the clinical importance of antimicrobial peptides (AMP) also increases. Up to now the possible use of AMPs is still experimental; however, individual AMPs are already established in the routine therapy (e. g. colistin). Further diagnostic and therapeutic measures may include photodynamic therapy, ultraviolet (UV) light application and differentiated genome analysis as well as the individual metabolism situation (metabolomics) of the pathogen cell and the patient tissue.

Neuropsychiatric disease relevance of circulating anti-NMDA receptor autoantibodies depends on blood-brain barrier integrity.

In 2007, a multifaceted syndrome, associated with anti-NMDA receptor autoantibodies (NMDAR-AB) of immunoglobulin-G isotype, has been described, which variably consists of psychosis, epilepsy, cognitive decline and extrapyramidal symptoms. Prevalence and significance of NMDAR-AB in complex neuropsychiatric disease versus health, however, have remained unclear. We tested sera of 2817 subjects (1325 healthy, 1081 schizophrenic, 263 Parkinson and 148 affective-disorder subjects) for presence of NMDAR-AB, conducted a genome-wide genetic association study, comparing AB carriers versus non-carriers, and assessed their influenza AB status. For mechanistic insight and documentation of AB functionality, in vivo experiments involving mice with deficient blood-brain barrier (ApoE(-/-)) and in vitro endocytosis assays in primary cortical neurons were performed. In 10.5% of subjects, NMDAR-AB (NR1 subunit) of any immunoglobulin isotype were detected, with no difference in seroprevalence, titer or in vitro functionality between patients and healthy controls. Administration of extracted human serum to mice influenced basal and MK-801-induced activity in the open field only in ApoE(-/-) mice injected with NMDAR-AB-positive serum but not in respective controls. Seropositive schizophrenic patients with a history of neurotrauma or birth complications, indicating an at least temporarily compromised blood-brain barrier, had more neurological abnormalities than seronegative patients with comparable history. A common genetic variant (rs524991, P=6.15E-08) as well as past influenza A (P=0.024) or B (P=0.006) infection were identified as predisposing factors for NMDAR-AB seropositivity. The >10% overall seroprevalence of NMDAR-AB of both healthy individuals and patients is unexpectedly high. Clinical significance, however, apparently depends on association with past or present perturbations of blood-brain barrier function.

Common variant at 16p11.2 conferring risk of psychosis.

Epidemiological and genetic data support the notion that schizophrenia and bipolar disorder share genetic risk factors. In our previous genome-wide association study, meta-analysis and follow-up (totaling as many as 18 206 cases and 42 536 controls), we identified four loci showing genome-wide significant association with schizophrenia. Here we consider a mixed schizophrenia and bipolar disorder (psychosis) phenotype (addition of 7469 bipolar disorder cases, 1535 schizophrenia cases, 333 other psychosis cases, 808 unaffected family members and 46 160 controls). Combined analysis reveals a novel variant at 16p11.2 showing genome-wide significant association (rs4583255[T]; odds ratio=1.08; P=6.6 × 10(-11)). The new variant is located within a 593-kb region that substantially increases risk of psychosis when duplicated. In line with the association of the duplication with reduced body mass index (BMI), rs4583255[T] is also associated with lower BMI (P=0.0039 in the public GIANT consortium data set; P=0.00047 in 22 651 additional Icelanders).

Ebstein's anomaly may be caused by mutations in the sarcomere protein gene MYH7.

Ebstein's anomaly is a rare congenital heart malformation characterised by adherence of the septal and posterior leaflets of the tricuspid valve to the underlying myocardium. Associated abnormalities of left ventricular morphology and function including left ventricular noncompaction (LVNC) have been observed. An association between Ebstein's anomaly with LVNC and mutations in the sarcomeric protein gene MYH7, encoding ß-myosin heavy chain, has been shown by recent studies. This might represent a specific subtype of Ebstein's anomaly with a Mendelian inheritance pattern. In this review we discuss the association of MYH7 mutations with Ebstein's anomaly and LVNC and its implications for the clinical care for patients and their family members.

Linking Patient-Centered Outcomes to Neuropsychological Practice: A Conceptual Framework and Opportunities for Research.

In the current healthcare climate, reimbursement for services is increasingly linked to the ability to demonstrate beneficial patient outcomes. Neuropsychology faces some unique challenges in outcomes research, namely, that neuropsychologists often do not follow patients over time and the effect of neuropsychological services on patient outcomes may not be fully realized until under another provider's care. Yet there is an urgent need for empirical evidence linking neuropsychological practice to positive patient outcomes. To provide a framework for this research, we define a core set of patient-centered outcomes and neuropsychological processes that apply across practice settings and patient populations. Within each area, we review the available existing literature on neuropsychological outcomes, identifying substantial gaps in the literature for future research. This work will be critical for the field to demonstrate the benefit of neuropsychological services, to continue to advocate effectively for reimbursement, and to ensure high-quality patient care.

Positive and negative bioimprinted polymeric substrates: new platforms for cell culture.

Bioimprinting, which involves capturing cell morphological details into a polymer matrix, provides a new class of patterned surfaces which opens an opportunity to investigate how cells respond to their own signatures and may introduce possibilities for regulating their behaviour. In this study, phenotypic details of human nasal chondrocytes (HNCs) were replicated in soft polydimethylsiloxane (PDMS) mould resulting in inverse replicas of cells, which have been termed here as 'negative bioimprint'. For the first time, the information from this negative bioimprint was then transferred into another PDMS layer resulting in surfaces which resemble cell morphology and were called 'positive bioimprints'. Soft lithography was used to transfer these details from PDMS into different polymers like polystyrene, tissue culture polystyrene and clinically used block co-polymer poly (ethylene glycol) terephthalate-poly (butylene terephthalate) (PEGT-PBT). Results obtained from surface characterization confirmed that fine details of cells were successfully replicated from cells to different polymer matrices without any significant loss of information during the different steps of pattern transfer. HNCs seeded on different polymer surfaces with positive and negative bioimprints exhibited distinct behaviour. Cells cultured on positive bioimprints were more spread out and displayed high levels of proliferation compared to those on negative bioimprints, where cells were more compact with lower proliferation.

Electron microscopy of cultured human corneas. Osmotic hydration and the use of a dextran fraction (dextran T 500) in organ culture.

Human corneas were studied by means of electron microscopy after culture at 31 degrees C for two to 20 days in a medium containing 8% dextran T 500. Dextran T 500, a strong osmotic agent, was included in the culture medium to prevent excessive swelling of the cornea. In order to exclude the possibility that the observed effects were the result of osmotic changes during fixation, in each experiment, fixatives with different osmolalities were used (430, 574, 727, and 812 mOsm). After 8, 16, and 20 days of culture, vacuoles appeared that were filled with dextran; the cytoplasm was completely filled with these vacuoles. The vacuolization was not limited to the endothelium, but was also observed in stromal keratocytes and to a limited extent in the epithelium. It was concluded that the monolayer of endothelium was still intact after 20 days of culture in medium containing 8% dextran T 500, and, secondly, that the dextran might have been taken up by endocytosis. Whether or not the uptake of dextran has a long-term toxic effect on the corneal cells remains to be elucidated.

Assessment of endothelial cell density in bovine corneas after osmotically induced dilation of intercellular spaces.

Corneal endothelial cells were visualized in intact bovine eyes by specular microscopy of unstained and stained cell borders. The corneas were excised, and flat corneal preparations were studied by reflected and transmitted light. In the excised corneas, the cell borders were visualized by osmotically induced dilation of intercellular spaces, alizarin red staining, and a combination of alizarin red and trypan blue staining. In whole eyes and in excised corneas, the estimates of endothelial cell densities varied by less than 3% from one method of visualization to the next. Estimates of endothelial cell densities obtained in intact eyes at 1 mm Hg were highly correlated to, but 13.6% lower than, estimates in excised corneas. Estimates of endothelial cell density obtained at intraocular pressures of 50 mm Hg were 7.7% lower than estimates obtained at 1 mm Hg.

Autoshaping with common and distinctive stimulus elements, compact and dispersed arrays.

Four groups of pigeons were trained with a standard autoshaping procedure in which a brief fixed-duration interval always followed by a grain delivery alternated with a longer variable-duration interval never associated with grain delivery. One of two stimuli was always presented during each interval. One of them contained three black dots and a black star on a green background; the other contained four black dots on a green background. The four elements of each stimulus were arranged in a more compact array for two groups and in a more dispersed array for the other two groups. Which of the two stimuli preceded grain delivery was counterbalanced within each pair of groups. The speed of occurrence of the first autoshaped peck was not affected by whether the stimulus containing the distinctive star element preceded grain delivery, but autoshaping was faster when the stimulus arrays were compact than when they were dispersed. During 560 response-independent training trials that followed the first autoshaped peck, this pattern reversed; both discriminative control over responding and the relative frequency of pecking the stimulus that preceded grain delivery were greater for the two groups where this stimulus contained the discriminative element than for the two groups where it contained only common elements. During subsequent testing with stimuli containing only a single element each, the distinctive feature was responded to proportionately more often by the two groups for which it had been an element of the stimulus preceding grain delivery than by the two groups for which it had been an element of the stimulus complex that never was associated with grain delivery. These data add further support to the hypothesis that the initial occurrence of autoshaped responding and its subsequent maintenance are not affected by the same variables. They also suggest that automaintenance is as sensitive as response-dependent training to the presence or absence of a distinctive stimulus element among several common elements and that this sensitivity appears to be independent of the specific method used for presenting the stimuli during automaintenance.

Stimulus generalization from feeder to response key in the acquisition of autoshaped pecking.

During autoshaping, a 6-second presentation of one stimulus and a variable time 30-second presentation of a second stimulus alternated in appearance on a pigeon key. Grain always was delivered for 3 seconds at the end of the first stimulus interval. In the first experiment, autoshaped pecking of the stimulus preceding grain delivery began much sooner when that stimulus was a black vertical line on a white background and the other stimulus was green than when the opposite stimulus arrangement was used. Because these two stimuli differed in form, hue, brightness, and similarity in hue and brightness to the illumination of the raised feeder, three subsequent experiments examined whether the differential speed of autoshaping in the two groups was due to a feature-positive, feature-negative effect, a preference for brighter over darker stimuli, a simple preference for white over green, or stimulus generalization from the brightness or hue of the illuminated, raised feeder to the stimulus on the key preceding grain delivery. The data from these experiments showed that the first autoshaped key peck was most likely to be made to the stimulus of the same hue as that illuminating the feeder, regardless of whether that stimulus was positively or negatively associated with grain delivery. At least under some conditions, therefore, stimulus-generalization mediated response transfer of pecking grain in the presence of the hue illuminating the feeder to pecking the key illuminated by a similar hue appears to account for the occurrence of autoshaped key pecking.

Molar incisor hypomineralisation in a group of children and adolescents living in Dresden (Germany).

AIM: This was to determine the prevalence of Molar Incisor Hypomineralisation (MIH) and to evaluate possible causes of these enamel defects. METHODS: The study group consisted of 2,408 10-17 year old children born during 1985-1992 and living in Dresden (Germany). Enamel defects were recorded using the modified DDE index. Children with enamel defects and their parents were invited for a re-examination to record the medical history of the children during the first 3 years of life; these children were matched by age with other children with apparently normal first molars (control group). RESULTS: 135 (5.6%) of the children had demarcated opacities in at least one first molar, i.e. MIH. A significantly higher prevalence of MIH was seen in children born between 1989 and 1991 compared with those born before and after that period (p<0.01). The number of children returning for the medical history questionnaire was low, 31 out of 135 responded (test group). Although there were no significant differences between the test and control groups in terms of peri and neonatal complications or other health problems, the low return precluded any definitive interpretation. CONCLUSION: The overall prevalence of MIH in this study was low by comparison with other previous epidemiological reports.

A single gene defect causing claustrophobia.

Claustrophobia, the well-known fear of being trapped in narrow/closed spaces, is often considered a conditioned response to traumatic experience. Surprisingly, we found that mutations affecting a single gene, encoding a stress-regulated neuronal protein, can cause claustrophobia. Gpm6a-deficient mice develop normally and lack obvious behavioral abnormalities. However, when mildly stressed by single-housing, these mice develop a striking claustrophobia-like phenotype, which is not inducible in wild-type controls, even by severe stress. The human GPM6A gene is located on chromosome 4q32-q34, a region linked to panic disorder. Sequence analysis of 115 claustrophobic and non-claustrophobic subjects identified nine variants in the noncoding region of the gene that are more frequent in affected individuals (P=0.028). One variant in the 3'untranslated region was linked to claustrophobia in two small pedigrees. This mutant mRNA is functional but cannot be silenced by neuronal miR124 derived itself from a stress-regulated transcript. We suggest that loosing dynamic regulation of neuronal GPM6A expression poses a genetic risk for claustrophobia.

Dissociation of accumulated genetic risk and disease severity in patients with schizophrenia.

Genotype-phenotype correlations of common monogenic diseases revealed that the degree of deviation of mutant genes from wild-type structure and function often predicts disease onset and severity. In complex disorders such as schizophrenia, the overall genetic risk is still often >50% but genotype-phenotype relationships are unclear. Recent genome-wide association studies (GWAS) replicated a risk for several single-nucleotide polymorphisms (SNPs) regarding the endpoint diagnosis of schizophrenia. The biological relevance of these SNPs, however, for phenotypes or severity of schizophrenia has remained obscure. We hypothesized that the GWAS 'top-10' should as single markers, but even more so upon their accumulation, display associations with lead features of schizophrenia, namely positive and negative symptoms, cognitive deficits and neurological signs (including catatonia), and/or with age of onset of the disease prodrome as developmental readout and predictor of disease severity. For testing this hypothesis, we took an approach complementary to GWAS, and performed a phenotype-based genetic association study (PGAS). We utilized the to our knowledge worldwide largest phenotypical database of schizophrenic patients (n>1000), the GRAS (Göttingen Research Association for Schizophrenia) Data Collection. We found that the 'top-10' GWAS-identified risk SNPs neither as single markers nor when explored in the sense of a cumulative genetic risk, have any predictive value for disease onset or severity in the schizophrenic patients, as demonstrated across all core symptoms. We conclude that GWAS does not extract disease genes of general significance in schizophrenia, but may yield, on a hypothesis-free basis, candidate genes relevant for defining disease subgroups.

Human corneal endothelium in organ culture. The influence of temperature and medium of incubation.

Forty-four human corneas from patients between 21 and 86 years were incubated in Eagle's minimum essential medium with Earle's salts 10--46 h post mortem. The influence of incubation temperature and composition of the medium on endothelial survival was evaluated. Whole corneas were stained by alizarine red. Recent cell loss was indicated by morphological alterations in the endothelial pattern. After 20--28 h of incubation minimum cell loss was found at 31 degrees C when 8% Dextrane-250 and 20% serum or 8% Dextrane-500 and 10% serum was added to the medium.

Endothelial cell density in donor corneas.

In 68 pairs of donor corneas the endothelial cells were visualized by provoked swelling of the cell borders in isotonic saline. In cornea from donors below 50 years of age numerical cell density was correlated to age. This was not the case in corneas from older donors. The cell densities in paired corneas were correlated. The cell density in the second cornea of a pair was found to be within x +/- 10.97% (+/-1.98SD) of the first cornea. The precision of this estimate could only be improved insignificantly by correction for age, sex, mean cell density or variation of the mean.

A simple apparatus for controlled rate corneal freezing II.

A corneal freezer without electrical or mechanical function is described. Cooling is achieved by partial immersion of the freezing unit in liquid nitrogen. The cooling rate is controlled by the heat content in the unit and the facility for heat removal from the unit. Corneas are frozen in aluminum vials. Corneas are thawed in less than 30 sec without risk of over heating, when the vials are placed on the metal core of another freezing unit at room temperature.

Cryopreservation of human cadaver corneas regenerated at 31 degrees C in a modified tissue culture medium.

Human corneas were obtained 2-82 h post mortem, cultured for 20-28 h, in a modified tissue culture medium, frozen at a controlled rate, and thawed rapidly. The thawed corneas were subjected to 20-28 h of additional culture. Immediately after thawing, a mean endothelial cell damage of 11% was indicated by trypane blue staining. The mean endothelial cell loss during the subsequent culture was 34%. This cell loss was not related to post mortem time, to donor age, to cell loss during the primary culture, or to endothelial cell density.

Early morphological changes in organ cultured human corneal endothelium.

Nineteen human cadaver corneas with few damaged endothelial cells were incubated under tissue culture conditions for time periods ranging from five min to 48 h. Morphological alterations of the endothelial cells were studied in whole wet mounts stained by alizarine red-alkohol-trypane blue and by scanning electron microscopy. Joint meetings of three cells are characteristic for normal corneal endothelium. After 15--60 min of incubation, damaged cells were expelled from the coherent cell sheet by expanding neighbouring cells. Joint meetings of 5--8 expanding cells were formed. After 24 h of incubation, joint meetings of four cells were the dominating morphological abnormality. Morphological changes during reduction of the numbers of cells in joint meetings are described.