Solid-state NMR sequential assignments of α-synuclein

Parkinson’s disease is amongst the most frequent and most devastating neurodegenerative diseases. It is tightly associated with the assembly of proteins into high-molecular weight protein species, which propagate between neurons in the central nervous system. The principal protein involved in this process is α-synuclein which is a structural component of the Lewy bodies observed in diseased brain. We here present the solid-state NMR sequential assignments of a new fibrillar form of this protein, the first one with a well-ordered and rigid N-terminal part.     

The horseradish tree,Moringa pterygosperma (Moringaceae)—A boon to Arid Lands?

The horseradish tree (Moringa pterygosperma,) is being introduced into drought-ridden lands to augment the local food and fodder supply. The tree grows up to 5 m per year. The foliage is high in calcium and has half the oxalates of amaranth. Seeds yield edible oil and the seed meal is used as fertilizer and as a coagulant to clarify turbid water. The philanthropic center, ECHO (Educational Concerns for Hunger Organization), North Fort Myers, Florida, receives many requests for seeds. A missionary in Mali wrote: “The seeds you sent arrived during the worst year of 14 years of dry weather. Only the moringa survived, and they have flourished. ”Another seed shipment resulted, after harvesting a crop, in 25 000 trees being planted by university students and faculty, around laborers’ houses in Maranhao, Brazil. The tree is not limited to tropical lowlands, but thrives at elevations of 800-1200 m in protected mountain areas of southern Mexico. The long-range effects of ingesting various parts of the tree as food or folkmedicine need study. Attention should be given to horticultural improvement, perhaps through hybridization with one or more related species now being compared with M. pterygosperma in India and Africa. ХРеНовое дерево, Moringa pterygosperma F. Gaertn. (Moringaceae), Дар сыхим землям. Хреновое дерево, Moringa pterygosperma, вводится в бездождивые землии чтоб умножить местное снабжение пищи и корма. Дерево растет до 5 метров в год. Листья содержат много кальцию и половину щавелев по сравнени#x044E; с амарантом. Семя дают съедобное масло и семеная мука употребляется как одобрение и как коагулант для очищение мутной воды. Филантропический цэнтэр ECHO (Educational Concern for Hunger Organization, North Fort Myers, Florida) полужает мното просьб чтоб получить семя. Один миссионер из Мали писал “Семя которые вы послали, прибыли в самый сухой год за 14 лет сухой погоды. Только моринга пережила и цвела. Другая отправка семен была совершена после получения урожая и 25000 деревьев были посажены студентами и учителями около домов рабочих в Маранхау, Брразилия. Это дерево не ограаничевается тропическим климатом и преуспевает на уровне 800 до 1200 метров в защищенных горных местах южной Мексики.     

LPA induces osteoblast differentiation through interplay of two receptors: LPA1 and LPA4

The bioactive phospholipid, lysophosphatidic acid (LPA), acting through at least five distinct receptors LPA1–LPA5, plays important roles in numerous biological processes. Here we report that LPA induces osteoblastic differentiation of human mesenchymal stem cells hMSC‐TERT. We find that hMSC‐TERT mostly express two LPA receptors, LPA1 and LPA4, and undergo osteoblastic differentiation in serum‐containing medium. Inhibition of LPA1 with Ki16425 completely abrogates osteogenesis, indicating that this process is mediated by LPA in the serum through activation of LPA1. In contrast to LPA1, down‐regulation of LPA4 expression with shRNA significantly increases osteogenesis, suggesting that this receptor normally exerts negative effects on differentiation. Mechanistically, we find that in hMSC‐TERT, LPA induces a rise in both cAMP and Ca²⁺. The rise in Ca²⁺ is completely abolished by Ki16425, whereas LPA‐mediated cAMP increase is not sensitive to Ki16425. To test if LPA signaling pathways controlling osteogenesis in vitro translate into animal physiology, we evaluated the bones of LPA4‐deficient mice. Consistent with the ability of LPA4 to inhibit osteoblastic differentiation of stem cells, LPA4‐deficient mice have increased trabecular bone volume, number, and thickness. J. Cell. Biochem. 109: 794–800, 2010. © 2010 Wiley‐Liss, Inc.     

Affective Priming Using Facial Expressions Modulates Liking for Abstract Art

We examined the influence of affective priming on the appreciation of abstract artworks using an evaluative priming task. Facial primes (showing happiness, disgust or no emotion) were presented under brief (Stimulus Onset Asynchrony, SOA = 20ms) and extended (SOA = 300ms) conditions. Differences in aesthetic liking for abstract paintings depending on the emotion expressed in the preceding primes provided a measure of the priming effect. The results showed that, for the extended SOA, artworks were liked more when preceded by happiness primes and less when preceded by disgust primes. Facial expressions of happiness, though not of disgust, exerted similar effects in the brief SOA condition. Subjective measures and a forced-choice task revealed no evidence of prime awareness in the suboptimal condition. Our results are congruent with findings showing that the affective transfer elicited by priming biases evaluative judgments, extending previous research to the domain of aesthetic appreciation.     

Hepatic Mitochondrial Function Analysis Using Needle Liver Biopsy Samples

Backgrounds and Aim

Current assessment of pre-operative liver function relies upon biochemical blood tests and histology but these only indirectly measure liver function. Mitochondrial function (MF) analysis allows direct measurement of cellular metabolic function and may provide an additional index of hepatic health. Conventional MF analysis requires substantial tissue samples (>100 mg) obtained at open surgery. Here we report a method to assess MF using <3 mg of tissue obtained by a Tru-cut® biopsy needle making it suitable for percutaneous application.

Methods

An 18G Bard® Max-core® biopsy instrument was used to collect samples. The optimal Tru-cut® sample weight, stability in ice-cold University of Wisconsin solution, reproducibility and protocol utility was initially evaluated in Wistar rat livers then confirmed in human samples. MF was measured in saponin-permeabilized samples using high-resolution respirometry.

Results

The average mass of a single rat and human liver Tru-cut® biopsy was 5.60±0.30 and 5.16±0.15 mg, respectively (mean; standard error of mean). Two milligram of sample was found the lowest feasible mass for the MF assay. Tissue MF declined after 1 hour of cold storage. Six replicate measurements within rats and humans (n = 6 each) showed low coefficient of variation (<10%) in measurements of State-III respiration, electron transport chain (ETC) capacity and respiratory control ratio (RCR). Ischemic rat and human liver samples consistently showed lower State-III respiration, ETC capacity and RCR, compared to normal perfused liver samples.

Conclusion

Consistent measurement of liver MF and detection of derangement in a disease state was successfully demonstrated using less than half the tissue from a single Tru-cut® biopsy. Using this technique outpatient assessment of liver MF is now feasible, providing a new assay for the evaluation of hepatic function.     

Trends in cation, nitrogen, sulfate and hydrogen ion concentrations in precipitation in the United States and Europe from 1978 to 2010: a new look at an old problem

Industrial emissions of SO₂ and NO_x, resulting in the formation and deposition of sulfuric and nitric acids, affect the health of both terrestrial and aquatic ecosystems. Since the mid-late 20th century, legislation to control acid rain precursors in both Europe and the US has led to significant declines in both SO₄–S and H⁺ in precipitation and streams. However, several authors noted that declines in streamwater SO₄–S did not result in stoichiometric reductions in stream H⁺, and suggested that observed reductions in base cation inputs in precipitation could lessen the effect of air pollution control on improving stream pH. We examined long-term precipitation chemistry (1978–2010) from nearly 30 sites in the US and Europe that are variably affected by acid deposition and that have a variety of industrial and land-use histories to (1) quantify trends in SO₄–S, H⁺, NH₄–N, Ca, and NO₃–N, (2) assess stoichiometry between H⁺ and SO₄–S before and after 1990, and (3) examine regional synchrony of trends. We expected that although the overall efforts of developed countries to reduce air pollution and acid rain by the mid-late 20th century would tend to synchronize precipitation chemistry among regions, geographically varied patterns of fossil fuel use and pollution control measures would produce important asynchronies among European countries and the United States. We also expected that control of particulate versus gaseous emission, along with trends in NH₃ emissions, would be the two most significant factors affecting the stoichiometry between SO₄–S and H⁺. Relationships among H⁺, SO₄–S, NH₄–N, and cations differed markedly between the US and Europe. Controlling for SO₄–S levels, H⁺ in precipitation was significantly lower in Europe than in the US, because (1) alkaline dust loading from the Sahara/Sahel was greater in Europe than the US, and (2) emission of NH₃, which neutralizes acidity upon conversion to NH₄ ⁺, is generally significantly higher in Europe than in the US. Trends in SO₄–S and H⁺ in precipitation were close to stoichometric in the US throughout the period of record, but not in Europe, especially eastern Europe. Ca in precipitation declined significantly before, but not after 1990 in most of the US, but Ca declined in eastern Europe even after 1990. SO₄–S in precipitation was only weakly related to fossil fuel consumption. The stoichiometry of SO₄–S and H⁺ may be explained in part by emission controls, which varied over time and among regions. Control of particulate emissions reduces alkaline particles that neutralize acid precursors as well as S-containing particulates, reducing SO₄–S and Ca more steeply than H⁺, consistent with trends in the northeastern US and Europe before 1990. In contrast, control of gaseous SO₂ emissions results in a stoichiometric relationship between SO₄–S and H⁺, consistent with trends in the US and many western European countries, especially after 1991. However, in many European countries, declining NH₃ emissions contributed to the lack of stoichiometry between SO₄–S and H⁺.Recent reductions in NO_x emissions have also contributed to declines in H⁺ in precipitation. Future changes in precipitation acidity are likely to depend on multiple factors including trends in NO_x and NH₃ emission controls, naturally occurring dust, and fossil fuel use, with significant implications for the health of both terrestrial and aquatic ecosystems.     

Enrichment of adeno-associated virus serotype 5 full capsids by anion exchange chromatography with dual salt elution gradients

Adeno-associated virus-based gene therapies have demonstrated substantial therapeutic benefit for the treatment of genetic disorders. In manufacturing processes, viral capsids are produced with and without the encapsidated gene of interest. Capsids devoid of the gene of interest, or “empty” capsids, represent a product-related impurity. As a result, a robust and scalable method to enrich full capsids is crucial to provide patients with as much potentially active product as possible. Anion exchange chromatography has emerged as a highly utilized method for full capsid enrichment across many serotypes due to its ease of use, robustness, and scalability. However, achieving sufficient resolution between the full and empty capsids is not trivial. In this work, anion exchange chromatography was used to achieve empty and full capsid resolution for adeno-associated virus serotype 5. A salt gradient screen of multiple salts with varied valency and Hofmeister series properties was performed to determine optimal peak resolution and aggregate reduction. Dual salt effects were evaluated on the same product and process attributes to identify any synergies with the use of mixed ion gradients. The modified process provided as high as ≥75% AAV5 full capsids (≥3-fold enrichment based on the percent full in the feed stream) with near baseline separation of empty capsids and achieved an overall vector genome step yield of >65%.     

Causes of Morbidity and Mortality in Free-Living Birds in an Urban Environment in Germany

Free-living birds are often presented to veterinarians at rehabilitation centers as well as in private practice. Information about disease processes and causes of death of indigenous free-living birds can aid the clinician in establishing proper treatment and in the assessment of potential zoonotic risks. For the present study, pathogens as causes of morbidity and mortality were determined by performing a complete necropsy on free-living birds presented to the Clinic for Birds and Reptiles of the University of Leipzig (Germany) that died shortly after admission or were euthanized due to an unfavorable prognosis. Over a 2-year period, 251 birds representing 13 families (Accipitridae, Apodidae, Columbidae, Corvidae, Falconidae, Fringillidae, Hirundinidae, Paridae, Passeridae, Picidae, Strigidae, Sturnidae and Turdidae) were examined. Trauma (62%), including several bite injuries inducing bacterial septicemia caused by Pasteurella multocida, was the most common cause of morbidity. Parasitic disease (18%) was mainly caused by Trichomonas gallinae, Eucoleus dispar and Syngamus trachea. Metabolic disease (13%), including fibrous osteodystrophy, was almost exclusively limited to juvenile specimens. Different Enterobacteriaceae including E. coli, Salmonella Typhimurium DT040 as well as Mycobacterium avium ssp. avium were identified as causal agents of primary bacterial disease (5%). Primary bacterial infection as cause of death or disease was of major importance in nestlings. Viral infections, mycoses and intoxication had minor significance as causes of morbidity.     

<i>In vitro</i> Antibacterial Activity of <i>Moringa oleifera</i> Ethanolic Extract against Tomato Phytopathogenic Bacteria

The tomato (Solanum lycopersicum L.) is one of the world’s most important vegetable crops. Still, phytopathogenic bacteria affect the yield and quality of tomato cultivation, like Agrobacterium tumefeciens (At), Clavibacter michiganensis subsp. michiganensis (Cmm), Pseudomonas syringae pv. tomato (Pst), Ralstonia solanacearum (Rs), and Xanthomonas axonopodis (Xa). Synthetic chemical products are used mostly on disease plant control, but overuse generates resistance to bacterial control. This study aimed to evaluate the in vitro antibacterial activity of the ethanolic extract of Moringa oleifera Lam. leaves against At, Cmm, Pst, Rs, and Xa, as well as information about this plant species’ chemical composition. Antibacterial activity against pathogens observed by microplate technique, phytochemical screening, and FTIR analysis revealed different bio-active compounds on ethanolic extracts with antibacterial activity. The growth inhibition rate ranged between 0.08% and 99.94%. The inhibitory concentration, IC50, required to inhibit 50% of At, Cmm, Pst, Rs, and Xa bacterial growth, was 276.67, 350.48, 277.85, 351.49, and 283.22 mg/L, respectively. Inhibition of phytopathogen bacteria’s growth increased as the concentrations of the extract also increased. Moringa oleifera extract can be recommended as a potent bio-bactericide.     

The peculiarities of succinic acid production from rapeseed oil by Yarrowia lipolytica yeast

The process of succinic acid (SA) production represents the combination of microbial synthesis of α-ketoglutaric acid from rapeseed oil by yeast Yarrowia lipolytica VKM Y-2412 and subsequent decarboxylation of α-ketoglutaric acid by hydrogen peroxide to SA that leads to the production of 69.0 g l^?1 of SA and 1.36 g l^?1 of acetic acid. SA was isolated from the culture broth filtrate in a crystalline form. The SA recovery from the culture filtrate has certain difficulties due to the presence of residual triglycerides of rapeseed oil. The effect of different methods of the culture filtrate treatment and various sorption materials on the coagulation of triglycerides was studied, and as a result, the precipitation of residual triglycerides by acetone was chosen. The subsequent isolation procedures involved the decomposition of H₂O₂ in the filtrate followed by filtrate bleaching and acidification with a mineral acid, evaporation of filtrate, and SA extraction with ethanol from the residue. The purity of crystalline SA isolated from the culture broth filtrate achieved 97.6–100 %. The product yield varied from 62.6 to 71.6 % depending on the acidity of the supernatant.