Trehalose loading through the mitochondrial permeability transition pore enhances desiccation tolerance in rat liver mitochondria

AbstractTrehalose has extensively been used to improve the desiccation tolerance of mammalian cells. To test whether trehalose improves desiccation tolerance of mammalian mitochondria, we introduced trehalose into the matrix of isolated rat liver mitochondria by reversibly permeabilizing the inner membrane using the mitochondrial permeability transition pore (MPTP). Measurement of the trehalose concentration inside mitochondria using high performance liquid chromatography showed that the sugar permeated rapidly into the matrix upon opening the MPTP. The concentration of intra-matrix trehalose reached 0.29 mmol/mg protein (∼190 mM) in 5 min. Mitochondria, with and without trehalose loaded into the matrix, were desiccated in a buffer containing 0.25 M trehalose by diffusive drying. After re-hydration, the inner membrane integrity was assessed by measurement of mitochondrial membrane potential with the fluorescent probe JC-1. The results showed that following drying to similar water contents, the mitochondria loaded with trehalose had significantly higher inner membrane integrity than those without trehalose loading. These findings suggest the presence of trehalose in the mitochondrial matrix affords improved desiccation tolerance to the isolated mitochondria

Leviathan on trial: Should states be held criminally responsible?

AbstractMany political theorists, philosophers, and International Relations scholars argue that states are ‘corporate moral agents’, which can be held responsible in many of the same ways as individual moral agents. States can have debts, contractual obligations, reparative obligations, and duties. Should states also be subject to criminal responsibility and punishment? Thus far, the debate about state crime has focused on two general problems with corporate crime: whether corporate entities can have intentions (or mens rea); and whether it is possible to punish them. In this paper, I identify two problems with extending corporate criminal responsibility to the state. First, since there is no ‘international corporate law’ that regulates the internal structures of states, many states fail to meet the conditions for corporate agency (and hence for criminal responsibility). Second, since the most serious international crimes are not subject to a statute of limitations, the argument for state crime paves the way for forms of ‘historical punishment’ that few of its proponents would accept. Finally, I argue that it is unnecessary to hold states criminally responsible, and that state responsibility ought to be understood as reparative rather than punitive.Non

Effects of stabiliser usage and pasteurisation norm on some quality of long-life yoghurt

Bu çalışmada, iki farklı stabilizator (jelatin ve keçiboynuzu sakızı) ilave edilerek üretilen yoğurtlara iki farklı ısıl işlem uygulanmış (65$\circ$C' de 1 dakika ve 60$\circ$C' de 5 dakika) ve üretilen dayanıklı yoğurtların bazı fiziksel, kimyasal, mikrobiyolojik ve duyusal özellikleri depolamanın 1., 15., 30., 45. ve 60. günlerinde saptanmıştır. Uygulanan ısıl işlemler, yoğurdun fiziksel ve duyusal özellikleri ile aroma maddelerinin oluşumlarını olumsuz yönde etkilemiş, stabilizator olarak kullanılan jelatin fiziksel özellikleri olumlu yönde etkilemiş, keçiboynuzu sakızı sadece su salma oranını azaltmıştır (p0.05). Depolama süresince yoğurtların pH değerleri, viskoziteleri, asetaldehit ve uçucu yağ asitleri miktarları ile duyusal özelliklerinde görülen değişimlerin önemli düzeyde olduğu belirlenmiştir (p<0.05).In this research, two different temperature/time combinations (65$\circ$C/1 min, 60 $\circ$C/5 min) were applied to experimental yoghurt samples manufactured from cow's milk, contained antifungal agent and two different stabilizer (Gelatine, Locust Bean Gum) and the chemical, physical, microbiological and organoleptic properties of long-life yoghurts were investigated at 1 th, 15 th, 30 th, 45 th and 60 th days of storage. Physical properties and formation of aromatic compounds of yoghurts were negatively effected by applied different heat treatment, but gelatine effected this properties significantly (<0.05). Locust bean gum only decreased syneresis (<0.05). Heat treatment caused reduction of lactic acid bacteria and mould were not determined yoghurt samples. While organoleptic properties of yoghurts were significantly effected (<0.05), this effect of stabilizers were not determined. During storage, changing of pH values, viscosity, acetaldehyde, volatile fatty acids and organoleptic properties of long-life yoghurts were found to be at important level (<0.05)

Quantification of intracellular trehalose in wild-type CHO cells and CHO-TRET1 cells.

<p>Cells were incubated in fully complemented cell culture medium containing 400 mM trehalose for 4 hours (<i>n</i> = 3, ± SD).</p

Glass transition temperature of the dried spinning solution (1.8 M trehalose, 10 mM KCl, 5 mM glucose, 20 mM HEPES, 120 mM choline chloride, pH 7.4) using FTIR technique.

<p>The figure indicates a characteristic peak-position of ν-OH stretch plotted against decreasing temperature.</p

Survival of CHO-TRET1 cells spin-dried in solutions with or without trehalose, then stored in LN₂ for 1 h, and finally rehydrated.

<p>(A) Membrane integrity of spin-dried cells stored in LN₂ for 1 h and 45 min after thawing and rehydration (B) Micrograph of the spin-dried cells after thawing and rehydration. (C) Growth of spin-dried cells after thawing and rehydration. The values were normalized to the initial cell count (<i>n</i> = 10, ± SD).</p

Survival of CHO-TRET1 cells spin-dried in buffers with or without trehalose and rehydrated immediately following desiccation.

<p>(A) Membrane integrity of spin-dried cells 45 min after rehydration (B) Micrograph of the cell samples after spin drying and rehydration. (C) Growth of cells after spin-drying and rehydration. The values were normalized to the initial cell count (<i>n</i> = 10, ± SD).</p

FTIR analysis of the dried film obtained after spin drying of sample buffer (1.8 M trehalose, 10 mM KCl, 5 mM glucose, 20 mM HEPES, 120 mM choline chloride, pH 7.4).

<p>The ratio of I₁₆₅₀/I₁₁₅₀ was used to quantify local water contents.</p

Basic configuration of the spin-drying apparatus.

<p>The cells were grown on glass cover slips prior to the spin-drying. During spin-drying, the glass cover slip was held in place by a vacuum chuck.</p