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Biomedical subjects

R E Howells

Publications and source records attributed to R E Howells.

At least 91 records · Page 5Linked to original sources

The uptake in vitro of dyes, monosaccharides and amino acids by the filarial worm Brugia pahangi.

The uptake in vitro of various substances by Brugia pahangi was investigated using infective larvae obtained from Aedes aegypti and worms removed from Meriones unguiculatus at 2, 3, 10, 20 and 90 days post-infection. Worms incubated in growth medium 199 containing 1% Trypan blue possessed demonstrable dye in the oral orifice, the anterior oesophageal lumen and the external openings of the vulva and the cloaca or anus but the dye was not found in the gut lumen even after incubation for 24 h. No uptake of ferritin particles into the intestine of the worms was found and no fluorescence could be demonstrated in the gut lumen of worms incubated in medium containing 50% (v/v) fluorescein isothiocyanate-conjugated calf serum for up to 24 h. Trypan blue uptake by the gut of Aspiculuris tetraptera was clearly observed after incubation for several hours. The uptake of D-glucose and L-leucine by B. pahangi was demonstrated using autoradiographic and scintillation counting techniques and incorporation into worm tissues was detected. Glucose was found to be readily incorporated in the apical, glycogen-rich areas of the myocytes of worms of all ages studied and in the uterine epithelium of the adult female. In contrast, a lower incorporation of D-glucose was found in the eggs, embryos and vas deferens and especially in the gut. The incorporation of L-leucine occurred throughout the tissue of the worms during a 30 min incubation. Labelling was also located over the surface of the cuticle of the worms, when incubated for a period of 15 to 60 min in L-[H]leucine. Scintillation counting techniques demonstrated that there was no uptake of 14C-labelled L-glucose or sucrose by B. pahangi. The data presented on the uptake in vitro of nutrients or other compounds by infective larvae and adult stages of B. pahangi did not demonstrate an intestinal route of uptake but indicated that the transcuticular route of uptake may be employed.

Animals↗

The exsheathment of Brugia pahangi microfilariae under controlled conditions in vitro.

Two reproducible techniques for the exsheathment in vitro of microfilariae of Brugia pahangi, and other sheathed microfilariae, are described. Microfilariae were isolated from infected cat blood by filtration and suspended in Hank's Balanced Salt Solution. The first technique involved the incubation of isolated microfilariae for one hour in 20 mM CaCl2 in a phosphate-free Balanced Salt Solution, during which time approximately 90% of the microfilariae lost their sheaths. The second method of exsheathing microfilariae of B. pahangi involved exposure of microfilariae to solutions of endopeptidase (5.8 units/ml) or papaya extract protease (3.0 units/ml) in Ca2+-free HBSS. Exsheathment rates of 95--100% occurred within 30 minutes in both enzyme solutions. Both the Ca2+ ion and the endopeptidase technique have proven equally effective in stimulating exsheathment of microfilariae of Brugia malayi, Wuchereria bancrofti and Litomosoides carinii. Such artificially exsheathed microfilariae are used for in vitro cultivation studies. The viability of Ca2+- and endopeptidase-exsheathed microfilariae of B. pahangi has been confirmed by inoculation of exsheathed larvae into susceptible female mosquitoes.

Aedes↗

The chemotherapy of rodent malaria, XXVII. Studies on mefloquine (WR 142,490).

Mefloquine (WR 142,490) is a potent blood schizontocide active against drug-sensitive and drug-resistant lines of Plasmodium berghei. The ED50 and ED90 against the P. berghei N strain in albino mice are 1.5 and 3.8 mg/kg respectively. The highly chloroquine-resistant RC line is less sensitive and mefloquine is not fully effective at the maximum tolerated dose in the '4-day test'. Mefloquine has a similar mode of action to quinine both in vitro and (as demonstrated by the morphological changes it induces in P. berghei) in vivo, but is some 100 times more potent. Unlike quinine and WR 122,455 it appears not to interact with DNA. It has no causal prophylactic effect. Mixtures of mefloquine with pyrimethamine, sulphaphenazole or primaquine have an additive effect.

Animals↗

Schistosoma mansoni: tail loss in relation to permeability changes during cercaria-schistosomulum transformation.

The hind-body region of Schistosoma mansoni cercariae observed in the scanning electron microscope demonstrates various stages of contraction which may be compared with those of living larvae which are secreting the acetabular gland contents. No evidence for an extensive lesion was found in cercarial bodies which had shed their tails under experimental conditions. Experiments on the permeability of the larvae to sodium fluoride, methylene blue and amino acids demonstrated that tail loss significantly affects the permeability of the bodies although the effect is greater immediately after decaudation than at later times. Subsequent increases in permeability may be correlated with a change in the general body surface.

Amino Acids↗

Factors affecting surface changes in intact cercariae and cercarial bodies of Schistosoma mansoni.

The effect of different incubation media and of temperature on the induction of water sensitivity has been investigated in intact and tailless Schistosoma mansoni cercariae. Removal of the cercarial tail by vortex stirring and elevation of the temperature of the medium from 27 to 37 degrees C resulted in the rapid onset of permeability changes in the larvae. The rate of change was greater in water than in TC-199 or Hanks' BSS media. Lowering the pH of the medium or increasing the concentration of Ca2+ ions decreased the rate of permeability change: raising the pH of the medium or the addition of 10(-5) M EDTA enhanced the rate. Raising the temperature of the medium also increased the rate of permeability change in intact cercariae although the rates obtained varied with the different media tested, being greatest in TC-199. It is concluded that both temperature elevation and loss of the cercarial tail influence the onset and rate of permeability changes in cercarial bodies during the transformation to schistosomula.

Calcium Chloride↗