Proper paraffin slide storage is crucial for translational research projects involving immunohistochemistry stains.

The use of paraffin slides and tissue microarrays (TMA) is indispensable for translational research. However, storage of paraffin slides over time has a substantial detrimental effect on the quality and reliability of immunohistochemistry stains. Particularly affected by this issue may be any collaborative efforts where paraffin slides or TMAs are shipped to central laboratories and then 'biobanked' for some time until use. This article summarizes some of the key issues affecting loss of antigenicity on paraffin slides and some simple storage solutions to help maintain high quality immunohistochemistry results when paraffin slides must be stored for a certain time prior to use.

Rings um Jerusalem

von Hans Much

A Missense Change in the ATG4D Gene Links Aberrant Autophagy to a Neurodegenerative Vacuolar Storage Disease.

Inherited neurodegenerative disorders are debilitating diseases that occur across different species. We have performed clinical, pathological and genetic studies to characterize a novel canine neurodegenerative disease present in the Lagotto Romagnolo dog breed. Affected dogs suffer from progressive cerebellar ataxia, sometimes accompanied by episodic nystagmus and behavioral changes. Histological examination revealed unique pathological changes, including profound neuronal cytoplasmic vacuolization in the nervous system, as well as spheroid formation and cytoplasmic aggregation of vacuoles in secretory epithelial tissues and mesenchymal cells. Genetic analyses uncovered a missense change, c.1288G>A; p.A430T, in the autophagy-related ATG4D gene on canine chromosome 20 with a highly significant disease association (p = 3.8 x 10-136) in a cohort of more than 2300 Lagotto Romagnolo dogs. ATG4D encodes a poorly characterized cysteine protease belonging to the macroautophagy pathway. Accordingly, our histological analyses indicated altered autophagic flux in affected tissues. The knockdown of the zebrafish homologue atg4da resulted in a widespread developmental disturbance and neurodegeneration in the central nervous system. Our study describes a previously unknown canine neurological disease with particular pathological features and implicates the ATG4D protein as an important autophagy mediator in neuronal homeostasis. The canine phenotype serves as a model to delineate the disease-causing pathological mechanism(s) and ATG4D function, and can also be used to explore treatment options. Furthermore, our results reveal a novel candidate gene for human neurodegeneration and enable the development of a genetic test for veterinary diagnostic and breeding purposes.

Chemistry of pyrrocorphins: methylative opening of the macrocycle between rings A and D, a side reaction in the peripheral C-methylation of a 20-methyl-pyrrocorphinate

One of the minor products from the previously described peripheral -methylation of a magnesium()-20-methyl--pyrrocorphinate is a C-19-methylated 19,20-seco-corphinoid derivative which, on complexation with nickel() acetate, recyclizes to a nickel()-tetradehydro-corrinate.

Chemistry of pyrrocorphins: synthesis of isobacteriochlorins and pyrrocorphins bearing a methyl group at the meso position between rings A and D

A sigmatropic methyl shift from the angular position C-1 in ring to the position C-20 between rings and constitutes the crucial step in syntheses leading to a 20-methyl-isobacteriochlorin and to 20-methyl-pyrrocorphins which served as substrates in the investigation presented in the accompanying communication.

Are carbohydrate storage strategies of trees traceable by early–latewood carbon isotope differences?

Investigations of stable isotopes in early and latewood cellulose offer interesting insights to climate-driven adaptations of trees’ carbon storage strategy during different phenological phases.

Crop Yield Responses to Temperature Fluctuations in 19th Century Finland: Provincial Variation in Relation to Climate and Tree Rings

Past agricultural responses to climate variability can helps us to better understand the current and future impacts of climate change on agricultural production. We studied rye (Secale cereale) and barley (Hordeum vulgare) yield responses to temperature fluctuations in Finland during the period 1861–1913. Our analyses demonstrate the high sensitivity of non-industrialised northern agriculture to temperature anomalies. We found evidence of a strong relationship between monthly and seasonal mean temperatures and crop yields. In particular, high spring temperatures were associated with higher yields. Additionally, we tested temperature-sensitive tree-ring series for their value in indicating previous agricultural outputs. The results imply that tree-ring proxies (in particular, maximum latewood density) can provide novel material for studies of historical periods and locations where instrumentally measured climate and harvest data are not available.

Petition der Herren Schusterjungen Münchens an den Magistrat daselbst: Hoher und stellenweiser sehr vortrefflicher Magistrat! Während rings um uns herum diejenigen Bürger, welche sich in den großen Tagen des Februar und März ...

Welsch (Projektbearbeiter): Ätzende Satire von konservativ/reaktionärer Seite auf den Verlauf der Münchener Märzrevolution in Form einer Petition der Schusterjungen zwecks Anerkennung ihrer Verdienste

Alternative nighttime nutrition regimens in glycogen storage disease type I: a controlled crossover study

BACKGROUND Traditional approaches for nighttime glycemic control in glycogen storage disease type I (GSDI) include continuous tube feeding, or ingestion of uncooked corn starch (CS) at bedtime. A modified corn starch (MCS) has been shown to prolong euglycemia in some patients. The aim of this study was to evaluate whether stable nighttime glucose control can be achieved with other types of slowly digested carbohydrates in adult GSDI patients. METHODS In this cross-over study, nocturnal glucose control and fasting times were assessed with three different nocturnal nutrition regimens in five patients, using continuous glucose monitoring (CGMS) in an outpatient everyday life setting. For each patient, continuous glucose profiles were measured after ingestion of (1) CS, (2) MCS or (3) a pasta meal at bedtime, during 5 to 6 consecutive nights for each regimen. RESULTS Stable nocturnal glucose control was achieved for all patients with a pasta meal, with a mean duration of glycemia >3.5 mmol/l of 7.6 h (range 5.7-10.8), and >4 mmol/l of 7 h (5.2-9.2), similar to CS and MCS. Fasting glucose before breakfast on workdays (after 7.1 ± 0.8 h) was not significantly different between the three interventions (CS 4.1 ± 0.5 mmol/l, MCS 4.6 ± 0.7 mmol/l, pasta 4.3 ± 0.9 mmol/l). During prolonged fasting on weekends, longer duration of normoglycemia was achieved with CS or MCS than with pasta. CONCLUSION Consumption of cooked pasta is a suitable and more palatable alternative to uncooked corn starch to achieve nighttime glucose control in adult patients with GSDI.

Stable carbon and oxygen isotopes in tree rings show physiological responses of Pericopsis elata to precipitation in the Congo Basin

In equatorial regions, where tree rings are less distinct or even absent, the response of forests to high-frequency climate variability is poorly understood. We measured stable carbon and oxygen isotopes in anatomically distinct, annual growth rings of four Pericopsis elata trees from a plantation in the Congo Basin, to assess their sensitivity to recorded changes in precipitation over the last 50 y. Our results suggest that oxygen isotopes have high common signal strength (EPS = 0.74), and respond to multi-annual precipitation variability at the regional scale, with low δ18O values (28–29‰) during wetter conditions (1960–1970). Conversely, δ13C are mostly related to growth variation, which in a light-demanding species are driven by competition for light. Differences in δ13C values between fast- and slow-growing trees (c. 2‰), result in low common signal strength (EPS = 0.37) and are driven by micro-site conditions rather than by climate. This study highlights the potential for understanding the causes of growth variation in P. elata as well as past hydroclimatic changes, in a climatically complex region characterized by a bimodal distribution in precipitation.

Chemical characterization and quantification of the brown algal storage compound laminarin

In search of a meaningful stress indicator for Fucus vesiculosus we found that the often used quantitative determination procedures for the polysaccharide laminarin (beta-1,3-glucan) result in different kind of problems, uncertainties and limitations. This chemical long-term storage form of carbon enables perennial brown algae in seasonally fluctuating ecosystems to uncouple growth from photosynthesis. Because of this high ecological relevance a reliable and precise method for determination and quantification of laminarin is needed. Therefore, a simple, cold water extraction method coupled to a new quantitative liquid chromatography-mass spectrometrical method (LC-MS) was developed. Laminarin was determined in nine out of twelve brown algal species, and its expected typical molar mass distribution of 2000-7000 Da was confirmed. Furthermore, laminarin consisted of a complex mixture of different chemical forms, since fifteen chemical laminarin species with distinct molecular weights were measured in nine species of brown algae. Laminarin concentrations in the algal tissues ranged from 0.03 to 0.86% dry weight (DW). The direct chemical characterization and quantification of laminarin by LC-MS represents a powerful method to verify the biochemical and ecological importance of laminarin for brown algae. Single individuals of Laminaria hyperborea, L. digitata, Saccharina latissima, F. serratus, F. vesiculosus, F. spiralis, Himanthalia elongata, Cystoseira tamariscifolia, Pelvetia canaliculata, Ascophyllum nodosum, Halidrys siliquosa and Dictyota dichotoma were collected in fall (18.11.2013) during spring low tide from the shore of Finavarra, Co. Clare, west coast of Ireland (53° 09' 25'' N, 09° 06' 58'' W). After sampling, the different algae were immediately transported to the lab, lyophilized and sent to the University of Rostock. Laminarin was extracted with cold ultrapure water from the algal samples. Before extraction they were ground to < 1 mm grain size with an analytical mill (Ika MF 10 Basic). The algal material (approx. 1.5 g DW) was extracted in ultrapure water (8 mL) on a shaker (250 rpm) for 5 h. After the addition of surplus ultrapure water (4 mL) and shaking manually, 1 mL of the sample was filter centrifuged (45 µm) at 14,000 rpm (Hettich Mikro 22 R). The slightly viscous supernatant was free of suspended material and converted into a microvial (300 µL) for further analysis. The extracts were analyzed using liquid chromatography-mass spectrometry (LC-MS) analysis (LTQ Velos Pro ion trap spectrometer with Accela HPLC, Thermo Scientific). Laminarin species were separated on a KinetexTM column (2.6 µm C18, 150 x 3 mm). The mobile phase was 90 % ultrapure water and 10 % acetonitrile, run isocratically at a flow rate of 0.2 mL min-1. MS was working in ESI negative ion mode in a mass range of 100 - 4000 amu. Glucose contents were determined after extraction using high-performance liquid chromatography (HPLC). Extracted samples were analyzed in an HPLC (SmartLine, Knauer GmbH) equipped with a SUPELCOGELTM Ca column (30 x 7,8 mm without preColumn) and RI-detector (S2300 PDA S2800). Water was used as eluent at a flow rate of 0.8 mL min-1 at 75 °C. Glucose was quantified by comparison of the retention time and peak area with standard solutions using ChromGate software. Mannitol was extracted from three subsamples of 10-20 mg powdered alga material (L. hyperborea, L. digitata, S. latissima, F. serratus, F. vesiculosus, F. spiralis, H. elongata, P. canaliculata, A. nodosum, H. siliquosa) and quantified, following the HPLC method described by Karsten et al. (1991). For analyzing carbon and nitrogen contents, dried algal material was ground to powder and three subsamples of 2 mg from each alga thalli were loaded and packed into tin cartridges (6×6×12 mm). The packages were combusted at 950 °C and the absolute contents of C and N were automatically quantified in an elemental analyzer (Elementar Vario EL III, Germany) using acetanilide as standard according to Verardo et al. (1990).

Concepción heroica Homérica en The Lord of the rings

Fil: Pepe de Suárez, Luz E. A..