Identification and Partial Purification of a Stage-Specific 33 kDa Mitochondrial Protein as the Alternative Oxidase of the Trypanosoma brucei brucei Bloodstream Trypomastigotes

ABSTRACT. The glycerophosphate oxidase (GPO), the unique terminal oxidase of bloodstream trypanosome (TAO), appears to be functionally similar to the alternative oxidases of some plants and higher fungi. Immunoblotting of mitochondrial proteins of bloodstream trypomastigotes of Trypanosoma brucei brucei with monoclonal or polyclonal antibodies to Sauromatum guttatum (voodoo lily) and Symplocarpus foetidus (skunk cabbage) alternative oxidases respectively revealed two proteins of about 33 kDa (p33) and 68 kDa (p68). These proteins are not present in procyclic trypomastigotes. Electrophoresis under rigorous denaturing conditions indicated p68 to be the dimer of p33. Indirect immunofluorescent studies of bloodstream and procyclic trypomastigotes with monoclonal antibody to plant alternative oxidase also showed the localization of 33 kDa protein in the mitochondria of the bloodstream trypomastigotes. The functional TAO activity could be solubilized efficiently from the mitochondrial membrane of the bloodstream trypomastigotes by 1% NP-40 or 10 mM lauryl maltoside. When fractionated by Superose 12 gel filtration chromatography, p33 was co-purified with the TAO enzymatic activity. The apparent molecular size of the active enzyme complex was found to be 160 kDa. Gradual disappearance of the 33 kDa protein and the TAO enzymatic activity were well correlated during in vitro differentiation of the bloodstream to procyclic trypomastigotes. This study implies that the net biosynthesis of p33, an essential subunit of TAO, is decreased during differentiation from bloodstream to procyclic trypomastigotes.     

Observations on the biology and nutritive value of the African giant snail Archachatina marginata

Food habits, growth rate, reproductive performance, carcass characteristics and nutritive value of the giant snail were investigated in the field and with captive snails. Food consumed included fresh plant material, livestock faeces and dead animals. Clutch sizes varied from six to twelve and incubation periods from 30 to 45 days. Weight gains of young snails of average weight 20 g fed on diets of single plant species ranged from 18% on Talinum triangulare leaves to 200% on pawpaw fruit. Best growth was shown on leaves of cassava and cocoyam, grain chaff of maize and fruit of pawpaw. Dressing-out percentage of snails of 200 g weight was 37.8%. Nutrient composition of the edible portion compared closely with that of domestic livestock. Nitrogen-free extractive content was high at 5% and fat low at 1.3%. Mineral analysis revealed a high iron content of 12.2 mg/100 g in the edible carcass. The experiments showed that the snail could thrive in captivity on a variety of locally produced feeds, many of them rejected as waste products. It shows great promise as a domestic animal capable of being reared with negligible capital outlay.     

Field observations on the African giant rat Cricetomys gambianus Waterhouse in southern Nigeria

The African giant rat Cricetomys gambianus Waterhouse is found in a wide variety of habitats in Ibadan district. These include an abandoned farm in a degraded deciduous forest, a zoological garden complex, Agodi; and a forest plantation of teak (Tectona grandis L.F.), human habitation and derived Guinea savanna. They located their burrows in the most heavily shaded parts of the habitats. Burrows were found mostly inside deserted termite mounds and amongst roots of trees probably to provide insulation from heat. They are nocturnal, omnivorous rodents, feeding on a wide variety of root crops, fruits, molluscs and insects. Fruits of the oil palm (Elaeis guineensis Jacq.) or their remains were found in all burrows. Information on habitat preferences, food habits, as well as that obtained from the dimensions and design of burrows which are reported in this paper later provided a basis for the design of cages and husbandry techniques for a programme of domestication of this rodent, which is currently being carried out at University of Ibadan, Nigeria.     

Proacrosome and acrosome of the spermatozoon in Acanthobdella peledina (Annelida: Clitellata)

Summary

Proacrosome and acrosome of the primitive leech Acanthobdella peledina are described by means of transmission electron microscopy. The proacrosome develops in early spermatids and has the shape of a pot-bellied urn with an opening towards the nucleus. Its wall is formed by a thin vesicle. In its interior, many sections of tubular structures are visible. This urn is seated atop a short, electron-dense tube. The resultant acrosome is unusually elongated, with a helically coiled acrosomal tube forming its base. Above the tube the thin acrosomal vesicle encloses a central space, within which is the acrosomal rod. The acrosomal structures clearly indicate a sister-group relationship to the Euhirudinea, but do not corroborate the notion of close kinship with the Branchiobdellidae.     

The role of NADPH oxidase (NOX) enzymes in neurodegenerative disease

Recently, mounting evidence implicating reactive oxygen species (ROS) generated by NADPH oxidase (NOX) enzymes in the pathogenesis of several neurodegenerative diseases including Amyotrophic lateral sclerosis (ALS), Alzheimer’s (AD), Parkinson’s (PD) and polyglutamine disease, have arisen. NOX enzymes are transmembrane proteins and generate reactive oxygen species by transporting electrons across lipid membranes. Under normal healthy conditions, low levels of ROS produced by NOX enzymes have been shown to play a role in neuronal differentiation and synaptic plasticity. However, in chronic neurodegenerative diseases over-activation of NOX in neurons, as well as in astrocytes and microglia, has been linked to pathogenic processes such as oxidative stress, exitotoxicity and neuroinflammation. In this review, we summarize the current knowledge about NOX functions in the healthy central nervous system and especially the role of NOX enzymes in neurodegenerative disease processes.     

Behavioural changes in the African giant rat (Cricetomys gambianus Waterhouse) under domestication

A gradient of behavioural patterns was observed among the wild and the first four generations of the African giant rat. Escape reactions and agonistic behaviour were completely lost as from the second generation. Wild rats were easily tamed and those born in captivity were very social. However, being a burrowing rodent, preferring a cool environment in nature, they had a low tolerance to heat. Cannibalism was an important management problem which persisted to the fourth generation resulting in the mortality of 40% of the newly born. From the fifth generation onwards, rats became completely tame and docile. Cannibalism was reduced to less than 10% of the newly born at the fifteenth generation. At this stage, for all practical purposes the giant rat can be regarded as a domestic animal.