Production of hydroxyl radicals in contracting skeletal muscle of cats

Reactive oxygen species increase during exhaustive contraction of skeletal muscle, but characterization of the specific species involved and their rates of production during nonexhaustive muscle contraction have not been investigated. We hypothesized that the production rate of hydroxyl radical (.OH) increases in contracting muscle and that this rate is attenuated by pretreatment with deferoxamine (Def) or dimethylthiourea (DMTU). We measured the rate of production of .OH before, during, and after 5 min of intermittent static contraction of the triceps surae muscles in cats (n = 6) using the formation of p-, m-, and o-tyrosines by hydroxylation of phenylalanine. L-Phenylalanine (30 mg/kg i.v.) was administered to each animal 3 min before contraction. Blood samples were collected from the popliteal vein 1 min before contraction; 1, 3, and 4.5 min during contraction; and 1 min after contraction. During and after contraction, the cumulative production rates of p-, m-, and o-tyrosines were elevated by 42.84 +/- 5.41, 0.25 +/- 0.04, and 0.21 +/- 0.03 nmol.min-1.g-1, respectively, compared with noncontracting triceps surae muscles. Pretreatment with Def (10 mg/kg i.v.; n = 5) or DMTU (10 mg/kg i.v.; n = 4) decreased the cumulative rates of production of p-, m-, and o-tyrosines during and after contraction. Additionally, the rate of tyrosine production increased in proportion to the percentage of maximal tension developed by the triceps surae muscles. These results directly demonstrate that .OH is produced in vivo in the skeletal muscle of cats during intermittent static contraction and that production can occur before the onset of fatigue.     

Potassium uptake and water content in hepatocytes isolated from rat livers preserved in Euro-Collins and UW solutions and after transplantation

Isolated hepatocytes from the rat were used to assess the maintenance of liver cell function in relation to the composition of the preservation medium. After separation by collagenase, they were incubated in Krebs-Ringer-Bicarbonate medium (KRB), Euro-Collins (EC), or University of Wisconsin (UW) solutions. Potassium influx, cell volume, and transaminase release were measured in cells freshly separated from control livers or from livers preserved in vitro up to 12 h in these media or having undergone orthotopic liver transplantation (OLT). While ion exchange levels were retained in all media, cells shrank significantly in UW but were able to restore their volume after 3 h of liver preservation. With regard to in vivo conditions, UW appears to be the best medium to prevent edema and to maintain more stable potassium exchange and enzyme production. These results are of value for liver transplantation in humans.     

Apoptosis in the failing human heart

BACKGROUND: Loss of myocytes is an important mechanism in the development of cardiac failure of either ischemic or nonischemic origin. However, whether programmed cell death (apoptosis) is implicated in the terminal stages of heart failure is not known. We therefore studied the magnitude of myocyte apoptosis in patients with intractable congestive heart failure. METHODS: Myocardial samples were obtained from the hearts of 36 patients who underwent cardiac transplantation and from the hearts of 3 patients who died soon after myocardial infarction. Samples from 11 normal hearts were used as controls. Apoptosis was evaluated histochemically, biochemically, and by a combination of histochemical analysis and confocal microscopy. The expression of two proto-oncogenes that influence apoptosis, BCL2 and BAX, was also determined. RESULTS: Heart failure was characterized morphologically by a 232-fold increase in myocyte apoptosis and biochemically by DNA laddering (an indicator of apoptosis). The histochemical demonstration of DNA-strand breaks in myocyte nuclei was coupled with the documentation of chromatin condensation and fragmentation by confocal microscopy. All these findings reflect apoptosis of myocytes. The percentage of myocytes labeled with BCL2 (which protects cells against apoptosis) was 1.8 times as high in the hearts of patients with cardiac failure as in the normal hearts, whereas labeling with BAX (which promotes apoptosis) remained constant. The near doubling of the expression of BCL2 in the cardiac tissue of patients with heart failure was confirmed by Western blotting. CONCLUSIONS: Programmed death of myocytes occurs in the decompensated human heart in spite of the enhanced expression of BCL2; this phenomenon may contribute to the progression of cardiac dysfunction.     

Psychopharmacotherapy for addictive and comorbid disorders: current studies

Proper diagnosis of comorbid disorders is crucial in treatment planning for the dually diagnosed. Since psychoactive substance use can obfuscate the diagnosis, special care must be taken to exclude organically based syndromes. Adequate periods of abstinence should first be achieved and subsequently the patient re-examined for residual symptoms compatible with a nonaddictive, nonsubstance-induced psychiatric disorder. The integration of concurrent treatment of both the mental and the addictive disorders appears to be the best approach for treatment of comorbid psychiatric and addictive disorders. An abstinence-based model that typically utilizes a 12-step group therapy is often employed for the addictive illnesses. Other forms of psychosocial therapies such as case managers are being used as well. Presently, physicians' prescribing practices for comorbid addicted patients are based on traditional approaches to use of medications in psychiatric patients, and their attitudes towards addictive disorders may play a significant role in determining the overall success of treatment.     

Assessment of bog-body tissue preservation by pyrolysis-gas chromatography/mass spectrometry

Flash pyrolysis-gas chromatography/mass spectrometry (py-GC/MS) was used to assess the quality and mechanism of protein preservation in the tissue of Iron Age bog bodies from Lindow, UK, and south-eastern Drenthe, The Netherlands. Abundant pyrolysis products of the fresh skin tissue, including 2,5-diketopiperazines of Pro-Gly, Pro-Ala, Pro-Val, Pro-Pro and Hyp, were readily assigned to specific amino acid or dipeptide moieties. Comparison of the pyrolysates of the bog-body tissues with that of modern samples revealed qualitative similarities suggesting good preservation of the collagen and non-collagenous proteins in the ancient tissues. Examination of the pyrolysates of samples of fresh calf skin, which had been treated with various vegetable tanning agents, clearly revealed markers of non-hydrolysable tannins including 1,2-benzenediol, 1,3-benzenediol and 1,2,3-benzenetriol, although chromatographic quality inevitably diminished with increasing functionalization of the compounds. Such markers were not detected in the pyrolysates of the bog-body tissues. Instead 4-isopropenylphenol, a characteristic pyrolysis product of Sphagnum moss, was detected in both solvent-extracted and base-treated samples of tissue. The presence of 4-isopropenylphenol in the pyrolysates of the bog-body tissues provides evidence that their preservation involves reactions of amino acids with sphagnum acid, and possibly other agents derived from the peat. The study constitutes the first chemical characterization of the pyrolysis products of modern and ancient collagen.     

A Maximum Entropy Approach for Collaborative Filtering

Collaborative filtering (CF) involves predicting the preferences of a user for a set of items given partial knowledge of the user's preferences for other items, while leveraging a database of profiles for other users. CF has applications e.g. in predicting Web sites a person will visit and in recommending products. Fundamentally, CF is a pattern recognition task, but a formidable one, often involving a huge feature space, a large data set, and many missing features. Even more daunting is the fact that a CF inference engine must be capable of predicting any (user-selected) items, given any available set of partial knowledge on the user's other preferences. In other words, the model must be designed to solve any of a huge (combinatoric) set of possible inference tasks. CF techniques include memory-based, classification-based, and statistical modelling approaches. Among these, modelling approaches scale best with large data sets and are the most adept at handling missing features. The disadvantage of these methods lies in the statistical assumptions (e.g. feature independence), which may be unjustified. To address this shortcoming we propose a new model-based CF method, based on the maximum entropy principle. For the MS Web application, the new method is demonstrated to outperform a number of CF approaches, including naive Bayes and latent variable (cluster) models, support vector machines (SVMs), and the (Pearson) correlation method.     

The distribution of palladium in a Pd-modified 4130 steel

The addition of palladium to a 4130 steel reduces its sensitivity to stress corrosion cracking and to hydrogen-induced cracking. The atom probe field-ion microscope has been used to study the microstructure and microchemistry of these palladium-modified 4130 steels with particular emphasis on the distribution of palladium between the various phases and internal interfaces. The palladium was found to segregate to the ferrite-ferrite lath boundaries to form a mono-atomically thick adsorbate layer. Palladium-manganese precipitates both at the lath boundaries and in the ferrite matrix were also detected. Most of the palladium was found to remain in solution in the ferrite.     

Growth and characterization of GaAs films deposited on Ge/Si composite substrates by metalorganic chemical vapor deposition

We report the results of studies which have been made on heteroepitaxial layers of GaAs and AlGaAs grown by metalorganic chemical vapor deposition on composite substrates that consist of four different types of heteroepitaxial layered structures of Ge and Ge-Si grown by molecular beam epitaxy on (100)-oriented Si substrates. It is found that of the four structures studied, the preferred composite substrate is a single layer of Ge ∼1 μm thick grown directly on a Si buffer layer. The double-crystal X-ray rocking curves of 2 μm thick GaAs films grown on such substrates have FWHM values as small as 168 arc sec. Transmission electron micrographs of these Ge/Si composite substrates has shown that the number of dislocations in the Ge heteroepitaxial layer can be greatly reduced by an anneal at about 750° C for 30 min which is simultaneously carried out during the growth of the GaAs layer. The quality of the GaAs layers grown on these composite substrates can be greatly improved by the use of a five-period GaAs-GaAsP strained-layer superlattice (SLS). Using the results of these studies, low-threshold optically pumped AlGaAs-GaAs DH laser structures have been grown by MOCVD on MBE Ge/Si composite substrates.