Optimized Protocol for Proportionate CNS Cell Retrieval as a Versatile Platform for Cellular and Molecular Phenomapping in Aging and Neurodegeneration

Efficient purification of viable neural cells from the mature CNS has been historically challenging due to the heterogeneity of the inherent cell populations. Moreover, changes in cellular interconnections, membrane lipid and cholesterol compositions, compartment-specific biophysical properties, and intercellular space constituents demand technical adjustments for cell isolation at different stages of maturation and aging. Though such obstacles are addressed and partially overcome for embryonic premature and mature CNS tissues, procedural adaptations to an aged, progeroid, and degenerative CNS environment are underrepresented. Here, we describe a practical workflow for the acquisition and phenomapping of CNS neural cells at states of health, physiological and precocious aging, and genetically provoked neurodegeneration. Following recent, unprecedented evidence of post-mitotic cellular senescence (PoMiCS), the protocol appears suitable for such de novo characterization and phenotypic opposition to classical senescence. Technically, the protocol is rapid, efficient as for cellular yield and well preserves physiological cell proportions. It is suitable for a variety of downstream applications aiming at cell type-specific interrogations, including cell culture systems, Flow-FISH, flow cytometry/FACS, senescence studies, and retrieval of omic-scale DNA, RNA, and protein profiles. We expect suitability for transfer to other CNS targets and to a broad spectrum of engineered systems addressing aging, neurodegeneration, progeria, and senescence

Biomass removal promotes plant diversity after short-term de-intensification of managed grasslands.

Land-use intensification is one of the main drivers threatening biodiversity in managed grasslands. Despite multiple studies investigating the effect of different land-use components in driving changes in plant biodiversity, their effects are usually studied in isolation. Here, we establish a full factorial design crossing fertilization with a combined treatment of biomass removal, on 16 managed grasslands spanning a gradient in land-use intensity, across three regions in Germany. Specifically, we investigate the interactive effects of different land-use components on plant composition and diversity using structural equation modelling. We hypothesize that fertilization and biomass removal alter plant biodiversity, directly and indirectly, mediated through changes in light availability. We found that, direct and indirect effects of biomass removal on plant biodiversity were larger than effects of fertilization, yet significantly differed between season. Furthermore, we found that indirect effects of biomass removal on plant biodiversity were mediated through changes in light availability, but also by changes in soil moisture. Our analysis thus supports previous findings, that soil moisture may operate as an alternative indirect mechanism by which biomass removal may affect plant biodiversity. Most importantly, our findings highlight that in the short-term biomass removal can partly compensate the negative effects of fertilization on plant biodiversity in managed grasslands. By studying the interactive nature of different land-use drivers we advance our understanding of the complex mechanisms controlling plant biodiversity in managed grasslands, which ultimately may help to maintain higher levels of biodiversity in grassland ecosystems

Unilateral optic neuropathy following subdural hematoma: a case report

<p>Abstract</p> <p>Introduction</p> <p>Unilateral optic neuropathy is commonly due to a prechiasmatic affliction of the anterior visual pathway, while losses in visual hemifields result from the damage to brain hemispheres. Here we report the unusual case of a patient who suffered from acute optic neuropathy following hemispherical subdural hematoma. Although confirmed up to now only through necropsy studies, our case strongly suggests a local, microcirculatory deficit identified through magnetic resonance imaging <it>in vivo</it>.</p> <p>Case presentation</p> <p>A 70-year-old Caucasian German who developed a massive left hemispheric subdural hematoma under oral anticoagulation presented with acute, severe visual impairment on his left eye, which was noticed after surgical decompression. Neurologic and ophthalmologic examinations indicated sinistral optic neuropathy with visual acuity reduced nearly to amaurosis. Ocular pathology such as vitreous body hemorrhage, papilledema, and central retinal artery occlusion were excluded. An orbital lesion was ruled out by means of orbital magnetic resonance imaging. However, cerebral diffusion-weighted imaging and T2 maps of magnetic resonance imaging revealed a circumscribed ischemic lesion within the edematous, slightly herniated temporomesial lobe within the immediate vicinity of the affected optic nerve. Thus, the clinical course and morphologic magnetic resonance imaging findings suggest the occurrence of pressure-induced posterior ischemic optic neuropathy due to microcirculatory compromise.</p> <p>Conclusion</p> <p>Although lesions of the second cranial nerve following subdural hematoma have been reported individually, their pathogenesis was preferentially proposed from autopsy studies. Here we discuss a dual, pressure-induced and secondarily ischemic pathomechanism on the base of <it>in vivo </it>magnetic resonance imaging diagnostics which may remain unconsidered by computed tomography.</p

Kindliche Verhaltensauffälligkeiten im ersten Lebensjahr und mütterliche Belastung in der Zeit der COVID-19-Pandemie

Theoretischer Hintergrund: Die COVID-19-Pandemie stellte in vielen Bereichen eine Belastung für Familien dar, insbesondere durch die einschneidenden Beschränkungen zu Beginn. Fragestellung: Wie wirkte sich dies auf die Belastung von Müttern mit Kindern im ersten Lebensjahr und auf die kindliche Verhaltensregulation aus? Methode: In einer Online-Befragung schätzten 577 Mütter das Schrei-‍, Schlaf- und Fütter-/Essverhalten ihrer Kinder (M = 7.3; 0 – 12 Mon., SD = 3.25) und ihre eigene Belastung ein, zudem Stresserleben, depressive Symptomatik, Partnerschaftszufriedenheit und Bonding. Ergebnisse: Schlafprobleme traten bei 21.7 %, schwer tröstbares und häufiges Schreien bei 12.3 % und exzessives Schreien bei 1.6 % der Kinder auf. Mindestens jede fünfte Mutter fühlte sich durch Schreien oder Schlafprobleme belastet. Mehr Stress, beeinträchtigtes Bonding und weniger Partnerschaftszufriedenheit erklärten 17 – 21 % der Varianz der mütterlichen Belastung durch Schrei- und Schlafverhalten. In der Zeit der stärksten Beschränkungen zeigte sich signifikant mehr Belastung in der Gruppe von Müttern, die von vermehrtem Schreien und verlängerter Einschlaflatenz berichteten, sowie mit mindestens einem weiteren Kind im Haushalt (MANOVA). Diskussion und Schlussfolgerung: Pandemiebedingt belastend für Mütter im ersten Jahr scheinen eingeschränkter Zugang zum Versorgungssystem, die Betreuung von mehr als einem Kind sowie das Alter des Kindes zu sein, während eine gute Beziehung zum Kind (Bonding) und/oder zum Partner (Partnerschaftszufriedenheit) abmildernd wirken.Theoretical background: The COVID-19 pandemic placed a burden on families in several respects, particularly because of the severe confinement imposed at its beginning. The confinement in spring 2020 led to social disruption and a reduction of supportive structures. In the first year of an infant’s life, the psychological well-being of a mother–child dyad is particularly susceptible to external stressful changes. Research question: How did the restrictions from the pandemic affect families with children in the first year of the infant’s life, particularly infant regulatory problems and related maternal stress? Methods: In an online survey, N = 577 mothers reported on their infants’ behavior (0 – 12 months of age, M = 7.3 months, SD = 3.3) regarding their crying, sleeping, and feeding/eating behavior as well as the respective distress experienced during the restrictions. Measures of current maternal well-being included overall perceived stress, depressivity, relationship satisfaction, and maternal bonding. Frequency/duration of infantile crying, sleep latency as well as night awakenings and feeding/eating problems were surveyed following clinical criteria (DC: 0 – 5) and percentile scores, respectively. The association of infant behavior and maternal distress was examined using linear regression and MANOVA. Results: Overall, at least one in five mothers felt burdened by her child’s regulatory problems during the time of severe restrictions. More than one in four mothers reported being highly or very highly distressed by her child’s crying. Sleeping problems, such as prolonged sleep latency (> 90th percentile) or several nightly awakenings, were reported for 21.7 % of the children. Difficulty to console and frequent crying were reported for 12.3 % and excessive crying for 1.6 % of the children. Higher levels of stress as well as increased impairment in maternal bonding and less relationship satisfaction explained 17 – 21 % of the variance of maternal distress from crying and sleeping problems. Significantly more distress was evident in the group of mothers who reported increased crying and prolonged latency to fall asleep (> 45 min) and with more than one child in the household during the period of most severe restrictions (MANOVA). A lack of medical, psychotherapeutic, and other means of care was reported by nearly a quarter of the respondents and was the only restriction that was significantly related to the perception of more stressful child behavior. Discussion and conclusion: Pandemic-related stressors for first-year mothers appear to be enhanced by limited access to the care system, caring for more than one child, the increasing age of the child, while a good relationship with the child (bonding) and/or partner (relationship) satisfaction provide buffers

Models of <i>KPTN</i>-related disorder implicate mTOR signalling in cognitive and overgrowth phenotypes

KPTN-related disorder is an autosomal recessive disorder associated with germline variants in KPTN (previously known as kaptin), a component of the mTOR regulatory complex KICSTOR. To gain further insights into the pathogenesis of KPTN-related disorder, we analysed mouse knockout and human stem cell KPTN loss-of-function models. Kptn -/- mice display many of the key KPTN-related disorder phenotypes, including brain overgrowth, behavioural abnormalities, and cognitive deficits. By assessment of affected individuals, we have identified widespread cognitive deficits (n = 6) and postnatal onset of brain overgrowth (n = 19). By analysing head size data from their parents (n = 24), we have identified a previously unrecognized KPTN dosage-sensitivity, resulting in increased head circumference in heterozygous carriers of pathogenic KPTN variants. Molecular and structural analysis of Kptn-/- mice revealed pathological changes, including differences in brain size, shape and cell numbers primarily due to abnormal postnatal brain development. Both the mouse and differentiated induced pluripotent stem cell models of the disorder display transcriptional and biochemical evidence for altered mTOR pathway signalling, supporting the role of KPTN in regulating mTORC1. By treatment in our KPTN mouse model, we found that the increased mTOR signalling downstream of KPTN is rapamycin sensitive, highlighting possible therapeutic avenues with currently available mTOR inhibitors. These findings place KPTN-related disorder in the broader group of mTORC1-related disorders affecting brain structure, cognitive function and network integrity.</p

Framework and baseline examination of the German National Cohort (NAKO)

The German National Cohort (NAKO) is a multidisciplinary, population-based prospective cohort study that aims to investigate the causes of widespread diseases, identify risk factors and improve early detection and prevention of disease. Specifically, NAKO is designed to identify novel and better characterize established risk and protection factors for the development of cardiovascular diseases, cancer, diabetes, neurodegenerative and psychiatric diseases, musculoskeletal diseases, respiratory and infectious diseases in a random sample of the general population. Between 2014 and 2019, a total of 205,415 men and women aged 19–74 years were recruited and examined in 18 study centres in Germany. The baseline assessment included a face-to-face interview, self-administered questionnaires and a wide range of biomedical examinations. Biomaterials were collected from all participants including serum, EDTA plasma, buffy coats, RNA and erythrocytes, urine, saliva, nasal swabs and stool. In 56,971 participants, an intensified examination programme was implemented. Whole-body 3T magnetic resonance imaging was performed in 30,861 participants on dedicated scanners. NAKO collects follow-up information on incident diseases through a combination of active follow-up using self-report via written questionnaires at 2–3 year intervals and passive follow-up via record linkages. All study participants are invited for re-examinations at the study centres in 4–5 year intervals. Thereby, longitudinal information on changes in risk factor profiles and in vascular, cardiac, metabolic, neurocognitive, pulmonary and sensory function is collected. NAKO is a major resource for population-based epidemiology to identify new and tailored strategies for early detection, prediction, prevention and treatment of major diseases for the next 30 years. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s10654-022-00890-5