Tocopherol content in tissues of mammals of different ecogenesis

Concentration of vitamin E (α-tocopherol) was studied in liver, kidney, heart, and skeletal muscle of 12 mammalian species, of which nine inhabited under natural conditions, while three were farm-bred. In the majority of the species the highest tocopherol content has been found as a rule in the animal liver and kidney. Essential differences have been revealed in distribution and level of α-tocopherol including those in taxonomically close species. These differences seem to be due to ecological peculiarities of the animals. The obtained results allow considering α-tocopherol as one of factors of maintenance of the natural antioxidant system at the steady level.     

Expression of human N-myristoyltransferase inEscherichia coli. Comparison with N-myristoyltransferases expressed in different tissues

Myristoyl CoA:protein N-myristoyltransferase catalyzes the addition of myristate to the amino-terminal glycine residue of a number of eukaryotic proteins.Escherichia coli transformed with human NMT expression construct produced high levels of N-myristoyltransferase. Using the combination of ammonium sulfate precipitation, chromatography on SP-Sepharose fast flow and fast protein liquid chromatography on Mono-S, the enzyme was purified more than 100 fold with 40% yield. The hNMT fusion protein exhibited an apparent molecular weight of 53 kDa on SDS-polyacrylamide gel electrophoresis. Upon cleavage by the Enterokinase [(Asp)₄-Lys], the hNMT exhibited an apparent molecular mass of 49 kDa without loss of catalytic activity. The hNMT activity could be greatly activated severalfold with the use of Tris, SDS, ethanol and acetonitrile. The catalytic activity of hNMT was potently inhibited in a concentration dependent manner by NIP₇₁₁ a bovine brain NMT inhibitory protein with a half maximal inhibition of 31.0 nM. TheE. coli expressed hNMT was homogeneous and showed enzyme activity.Abbreviations NMT N-myristoyl CoA:protein N-myristoyltransferase - NIP₇₁ 71 kDa heat stable membrane bound N-myristoyltransferase inhibitor protein - hNMT human NMT - DTNB N-5,5dithiobis (2-nitrobenzoic acid) - FPLC fast protein liquid chromatography - IPTG isopropyl -D-thiogalactopyranoside - cDNA complementarydeoxyribonucleic acid - SDS sodium dodecyl sulfate - PMSF phenylmethylsulfonyl fluoride     

Increase in calcium content and Ca2+-ATPase activity in the brain of fasted rats: Comparison with different ages

The effect of fasting on calcium content and Ca²⁺-ATPase activity in the brain tissues of 5 weeks and 50 weeks old rats was investigated. Brain calcium content and Ca²⁺-ATPase activity in the microsomal and mitochondrial fractions of the brain homogenate from young and elderly rats were significantly increased by overnight–fasting. These increases were appreciably restored by a single oral administration of glucose solution (400 mg/100 g body weight) to fasted rats. In comparison with young and elderly rats, brain calcium content and microsomal Ca²⁺-ATPase activity were significantly elevated by increasing ages. The effect of ageing was not seen in the brain mitochondrial Ca²⁺-ATPase activity. When calcium (50 mg/100 g) was orally administered to young and elderly rats, brain calcium content was significantly elevated. The calcium administration–induced increase in brain calcium content was greater in elderly r crease in Ca²⁺-ATPase activity in the microsomal and mitochondrial fractions of brain homogenates from young rats. In aged rats, the microsomal Ca²⁺-ATPase activity was not further enhanced by calcium administration, although the mitochondrial enzyme activity was significantly raised. The present study demonstrates that the fasting–induced increase in brain calcium content is involved in Ca²⁺-ATPase activity raised in the brain microsomes and mitochondria of rats with different ages, supporting a energy–dependent mechanism in brain calcium accumulation.     

Characterization of methadone receptor subtypes present in human brain and lung tissues

In addition to their use in pain control, opioids can function as regulators of tumor cell growth. We have found that the therapeutic opioid, methadone, significantly inhibits the in vitro and in vivo growth of human lung cancer cells, and this effect appears to be mediated by specific, high affinity, non-conventional opioid binding sites. The present study indicates the existence of multiple subtypes of binding sites mediating the peripheral and central nervous system actions of this drug. Pharmacological and biochemical characterizations of the methadone binding sites expressed in human brain and normal lung tissues indicate that these sites are distinct from each other and from other opioid receptor types present on human and rat brain membranes, as well as those expressed in human lung cancer cells. The identification of distinct methadone receptor types in the different tissues could lead to the development of more selective and less toxic drugs targeted toward the tumor Cells.     

Comparison of glycopeptide in lung cancer tissues of different histological types

The glycopeptide was studied in the pulmonary cancer tissues of three different histological types including squamous cell carcinoma, undifferentiated cell carcinoma, and adenocarcinoma. The glycopeptide preparation was made by digesting the lung cancer tissues with proteolytic enzymes followed by removal of enzymes and undigested protein. The comparison of the relative proportion of the glycopeptide, which migrated toward the anode slower than hyaluronic acid, and was stained with both alcian blue and PAS, and contained higher sialic acid, was carried out among three different histologic types of lung cancer. This glycopeptide was highest in adenocarcinoma, followed by undifferentiated cell carcinoma, and was lowest in squamous cell carcinoma.     

Biomechanics of torsion in the human mandible

Comparative investigations of mandibular function among primates have relied upon elementary structural models to estimate states of masticatory stress and strain. In these studies, mandibular corpus morphology is idealized as a homogeneous, isotropic symmetrical body of invariant geometry, and this morphological abstraction is used to infer relative levels of stress and strain in the jaw. In reality, none of the limiting conditions assumed by these models is satisfied; consequently, it is prudent to ask whether this “textbook” engineering approach is valid for the inference of biomechanical behavior. In this study, the predictions of various geometric representations of the mandibular corpus are evaluated against strains recorded in a sample of human jaws loaded in torsion. Symmetrical geometrical models (including various “robusticity” shape indices), although convenient, are probably not consistently reliable for predicting the distribution of strains in the corpus. The experimental data suggest that variations in cortical thickness within sections play a significant role in determining the profile of relative strains. For comparative applications, characterization of the corpus as an asymmetrical hollow ellipse (i.e., with differing thickness of medial and lateral cortical plates) may provide a reasonable portrayal of relative strains. Am J Phys Anthropol 105:73–87, 1998. © 1998 Wiley-Liss, Inc.     

Comparison of plastid DNA replication in different cells and tissues of the rice plant

In a previous study, we mapped replication origin regions of the plastid DNA around the 3 end of the 23S rRNA gene in rice suspension-cultured cells. Here, we examined initiation of the plastid DNA replication in different rice cells by two-dimensional agarose gel electrophoresis. We show for the first time, to our knowledge, that the replication origin region of the plastid DNA differs among cultured cells, coleoptiles and mature leaves. In addition, digestion of the replication intermediates from the rice cultured cells with mung bean nuclease, a single-strand-specific nuclease, revealed that both two single strands of the double-stranded parental DNA were simultaneously replicated in the origin region. This was further confirmed by two-dimensional agarose gel analysis with single-stranded RNA probes. Thus, the mode of plastid DNA replication presented here differs from the unidirectional replication started by forming displacement loops (D-loops), in which the two D-loops on the opposite strands expand toward each other and only one parental strand serves as a template.     

Mass properties of the human mandible

Computer simulation of human masticatory dynamics requires specification of the jaw's mass properties. These are difficult to estimate, especially in living subjects. Here, we used calibrated computed tomography (CT) to determine the properties of eight osseous jaw specimens with adult dentitions. When the CT numbers were converted to mineral densities, the mean estimated jaw mass was 13% greater than the mean wet weight. Putative bone marrow accounted for an extra 7% of mass. The mean bone densities for the sample were very consistent (1.72+/-0.02g/cm(3)). The mass and geometric centers were close (mean linear difference 0.43+/-0.18mm), and were always located anteroposteriorly between the second and third molars. The largest moment of inertia (MI) occurred around the jaw's superoinferior axis, and the smallest around its transverse axis. Bone marrow added an extra 7% to the MIs. There were linear relationships between the mandibular length (expressed three dimensionally), the actual and estimated masses, and the moments of inertia. Our study suggests non-invasive imaging (such as magnetic resonance) and even direct linear measurement, may be adequate to estimate jaw mass properties in living humans.     

The cholesterol content of the human erythrocyte influences calcium influx through the channel

In order to study the influence of the cholesterol content on the calcium entry channel, the human red blood cell was used as a model system. The cholesterol to lecithin ratio (C/L ratio) of the membrane was modified experimentally by incubating the cells (15h, 25 degrees) with liposomes of defined C/L ratios. Subsequently, net 45Calcium-influx into the cell was measured by inhibiting the Ca-ejecting ATPase with vanadate. Additionally, the use of nitrendipine, a potent calcium channel inhibitor, during incubation allowed the determination of Ca-influx through the calcium channel. A positive correlation between the 45Ca++-influx and the molar C/L ratio of the membrane was found over a wide C/L range. A molar C/L ratio of 1.4 in the membrane increased calcium influx by 150 % compared to controls (molar C/L ratio = 0.8, calcium influx rate = 100 %), while a molar C/L ratio at less than 0.75 decreased calcium influx by 50 %. We conclude, that the cholesterol content of the membrane greatly influences the calcium channel and thus plays a pivotal role for the availability of calcium as a second messenger. These findings may provide a link between high plasma cholesterol and the development of atherosclerosis as well as enhanced platelet aggregability.     

Comparison of calcium accumulation between the arteries of human and monkey

To examine whether an accumulation of elements in the arteries was affected by the way of walking, the authors investigated age-related changes of elements in the arteries of the Japanese, Thai, and Japanese monkeys. After the ordinary dissections by medical students were finished, the subclavian, axillary, brachial, radial, common iliac, externaliliac, femoral, and posterior tibial arteries were resected from the subjects of the Japanese and Thai over 60 yr of age and they were also resected from the Japanese monkeys over 20 yr of age. The element content was determined by inductively coupled plasma-atomic emission spectrometry. It was found that in an comparison between the arteries of anatomically corresponding regions, the average content of Ca was eight times or four times higher in the arteries of the lower limb than in the arteries of the upper limb in the Japanese or Thai, over 60 yr of age, respectively. In the Japanese monkeys over 20 yr of age, the average content of Ca was 1.2 times higher in the arteries of the lower limb than in the arteries of the upper limb. The result suggests that an accumulation of Ca in the arteries of the lower limb with aging is affected by the way of walking.