Visualization of mitochondria with green fluorescent protein in cultured fibroblasts from patients with mitochondrial diseases

cDNAs for green fluorescent protein (GFP) and for a GFP fusion protein containing the presequence of human ornithine transcarbamylase (pOTC-GFP) were transfected into cultured human fibroblasts. GFP cDNA gave diffuse fluorescence throughout the cytoplasm and the nucleus, whereas pOTC-GFP cDNA gave mitochondria-associated fluorescence. Fluorescent mitochondrial structures could be classified into five patterns: thread-like mitochondria, fine thread-like ones, rod-like ones, granular ones, and granular ones with weak cytosolic fluorescence. pOTC-GFP mutants resulted in a loss of mitochondrial fluorescence and an appearance of weak fluorescence throughout the cytoplasm. pOTC-GFP cDNA was transfected into fibroblasts from patients with various mitochondrial diseases. Higher ratios of fibroblasts with granular mitochondria and those with fine thread-like ones were observed in a patient with Reye's syndrome and a patient with Kearns-Sayre syndrome. Weak cytosolic fluorescence was sometimes observed in fibroblasts from these patients. This method will be useful to analyze mitochondrial structural alterations and disorders of mitochondrial protein import.     

Malignant bone pain: pathophysiology and treatment

2-acetyl aminofluorene (AAF) reacts in acidic conditions with nitrous fume yielding N-nitroso-AAF (N-NO-AAF), as previously described, that exerts more toxic and mutagenic effects than its parental compound. In this study, the effect of sodium nitrite (NaNO2) on the tumorigenicity of AAF in rats fed with AAF and NaNO2 was observed. Wistar rats were divided into five groups: group I served as control; group II were treated with NaNO2 (0.3%); group III was given 0.02% AAF alone; groups IV and V received both AAF and NaNO2 (0.2 and 0.3% respectively) in their diet for 12 weeks. At the end of the experiment, all rats in groups III, IV and V developed early stage phenomena of hepatocellular carcinoma, including hepatomegaly with variable-sized foci and neoplastic nodules. Severe damage was observed in the rats treated with AAF and NaNO2. Feeding of AAF (0.02%) for 3 months elevated the levels of c-Fos, c-Jun and c-Myc proteins in the rat livers. The AAF-induced c-Jun, c-Fos and c-Myc expressions were significantly magnified (P < 0.001) by NaNO2. These data confirmed that the strengthening of AAF-induced hepatocarcinogenesis by NaNO2 should be associated with its enhancing effect on the AAF-induced increases in the expressions of c-Jun, c-Fos and c-Myc.     

Platelet-derived growth factor activates a mammalian Ste20 coupled mitogen-activated protein kinase in airway smooth muscle

Attempts to immunise sheep against natural infestations by Lucilia cuprina larvae have not been effective. Yet it is known that the larvae excrete the immunosuppressant ammonium bicarbonate. The effect of larval ammonium and nonionic ammonia on immunopathobiology was evaluated in 12 infested sheep. The concentration of ammonium in veins draining infested sites was measured in another group of four sheep. Mean jugular unionized ammonia concentration increased 3.5 to 5.6 times above pre-infested control levels. Mean venous ammonium concentrations draining infested sites were 13 times higher than pre-infested jugular or carotid levels. Increases in jugular ammonia concentrations correlated with increased number of larvae, area of infestation, earlier death, neutropenia, eosinopenia, lymphocytopenia, large declines in serum globulins and zinc, and large rises in toxic neutrophils. The high concentrations of toxic unionized ammonia in blood directly permanently damaged neutrophils and lymphocytes and depressed serum globulin production. The results show that the ammonium from the excreta of larvae of L. cuprina may be highly immunosuppressive.     

Error in the UNITY substitution rule for subscripted operators

UNITY, introduced by Chandy and Misra [ChM88], is a programming logic intended to reason about temporal properties of distributed programs. Despite the fact that UNITY does not have the full power of, for example, linear temporal logic, it enjoys popularity due to its simplicity.There was however a serious problem with the Substitution Rule. The logic is incomplete without the rule, and with the rule it is inconsistent.     

Prolog/Rex-a way to extend Prolog for better knowledgerepresentation

Prolog/Rex represents a powerful amalgamation of the latest techniques for knowledge representation and processing, rich in semantic features that ease the difficult task of encoding heterogeneous knowledge of real-world applications. The Prolog/Rex concept mechanism lets a user represent domain entities in terms of their structural and behavioral properties, including multiple inheritance, arbitrary user-defined relations among entities, annotated values (demons), incomplete knowledge, etc. A flexible rule language helps the knowledge engineer capture human expertise and provide flexible control of the reasoning process. Additional Prolog/Rex strength that cannot be found in any other hybrid language made on top of Prolog is language level support for keeping many potentially contradictory solutions to a problem, allowing possible solutions and their implications to be automatically generated and completely explored before they are committed. The same mechanism is used to model time-states, which are useful in planning and scheduling applications of Prolog/Rex     

Weight smoothing to improve network generalization

A weight smoothing algorithm is proposed in this paper to improve a neural network's generalization capability. The algorithm can be used when the data patterns to be classified are presented on an n-dimensional grid (n>/=1) and there exists some correlations among neighboring data points within a pattern. For a fully-interconnected feedforward net, no such correlation information is embedded into the architecture. Consequently, the correlations can only be extracted through sufficient amount of network training. With the proposed algorithm, a smoothing constraint is incorporated into the objective function of backpropagation to reflect the neighborhood correlations and to seek those solutions that have smooth connection weights. Experiments were performed on problems of waveform classification, multifont alphanumeric character recognition, and handwritten numeral recognition. The results indicate that (1) networks trained with the algorithm do have smooth connection weights, and (2) they generalize better.     

Network information criterion-determining the number of hiddenunits for an artificial neural network model

The problem of model selection, or determination of the number of hidden units, can be approached statistically, by generalizing Akaike's information criterion (AIC) to be applicable to unfaithful (i.e., unrealizable) models with general loss criteria including regularization terms. The relation between the training error and the generalization error is studied in terms of the number of the training examples and the complexity of a network which reduces to the number of parameters in the ordinary statistical theory of AIC. This relation leads to a new network information criterion which is useful for selecting the optimal network model based on a given training set.     

An algebraic theory of object-oriented systems

The paper presents an algebraic specification of net objects. By net objects we mean those that are defined in object-oriented paradigms and those defined with nested relations in complex database models. An algebraic specification is set up that involves structures of net objects, accesses of net objects, and some features of object-oriented programming, such as multiple inheritance and polymorphism. Objects and their relationships are then characterized formally in the modeling, which utilizes the hierarchical approach in the algebraic theory of abstract data types, and is further developed by adding mechanisms from existing object systems. Categories of net objects are presented with the properties of accesses among them     

Structuring fault-tolerant object systems for modularity in adistributed environment

The object-oriented approach to system structuring has found widespread acceptance among designers and developers of robust computing systems. The authors propose a system structure for distributed programming systems that support persistent objects and describe how properties such as persistence and recoverability can be implemented. The proposed structure is modular, permitting easy exploitation of any distributed computing facilities provided by the underlying system. An existing system constructed according to the principles espoused here is examined to illustrate the practical utility of the proposed approach to system structuring