Spontaneous sexual differentiation of Mesocestoides corti tetrathyridia in vitro

Barrett N. J., Smyth J. D. and Ong S. J. 1982. Spontaneous sexual differentiation of Mesocestoides corti tetrathyridia in vitro. International Journal for Parasitology12: 315–322. Tetrathyridia of Mesocestoides corti, from the body cavity of mice, maintained in the laboratory by intraperitoneal infection, were used for in vitro culture. In an initial experiment, after 50 days asexual multiplication in vitro one tetrathyridium spontaneously segmented and developed into a sexually mature adult. Further experiments were carried out in an attempt to determine the conditions favouring segmentation and sexual differentiation. A combination of 5 or 10 ml liquid medium S1OE.H (basically composed of CMRL 1066 and foetal calf serum with supplements) changed every 3 days, in a Leighton tube (19 × 105 mm), rotated at 38°C and gassed with 10 or 20% CO₂, containing between 100 and 200 tetrathyridia, has proved to be most suitable so far. Numerous adult worms with normal male and female genitalia have been obtained in this system. However, segmentation is sporadic, rather than consistent and only a few shelled eggs with hooked oncospheres have so far been obtained, suggesting that impregnation and fertilization in vitro is not fully comparable with that in vivo.     

Effects of certain lipid compounds on growth and asexual multiplication of Mesocestoides corti (Cestoda) tetrathyridia

The effects of Williams and Law mixture (W-L), farnesol (juvenile hormone mimics), ecdysterone (molting hormone analog), cholesterol, and stigmasterol on growth and asexual reproduction of Mesocestoides corti larvae in vivo and in vitro were studied. W-L and ecdysterone stimulated an increase in larval asexual reproduction under in vivo conditions, however, cholesterol had no effect on the larvae in vivo. In vitro, cholesterol stimulated a slight increase in reproduction but had no effect on growth; stigmasterol appeared to be somewhat detrimental to the larvae. In vitro, W-L, farnesol, and ecdysterone stimulated by varying amounts an increase in growth (as measured by larval size) and asexual reproduction. Lipid extraction of M. corti larvae and Hymenolepis diminuta adults, followed by a series of thin-layer chromatograms, yielded 2 compounds (I-B and I-D) which affected M. corti larvae in vitro. These 2 compounds stimulated increased growth of the M. corti larvae and to some extent an increase in asexual reproduction.     

Protection against Mesocestoides corti infection in mice treated with zymosan or Salmonella enteritidis 11RX

Toye P. G. and Jenkin C. R. 1982. Protection against Mesocestoides corti infection in mice treated with zymosan or Salmonella enteritidis 11RX. International Journal for Parasitology12: 399–402. Zymosan and Salmonella enteritidis 11RX were found to partially protect mice against infection with the cestode Mesocestoides corti. Thus, mice previously infected with S. enteritidis 11RX contained fewer parasites in the peritoneal cavity compared to normal mice. Mice pretreated with zymosan contained fewer parasites in the peritoneal cavity and in the liver compared to normal mice and this protection was enhanced by the passive transfer of serum from mice chronically infected with M. corti. Examination of mice in the initial stages of infection revealed that the administration of zymosan led to an alteration in parasite location from the peritoneal cavity to the liver.     

Cross-protection between the metacestodes of Mesocestoides corti and Taenia crassiceps in mice

Novak M. 1984. Cross-protection between the metacestodes of Mesocestoides corti and Taenia crassiceps in mice. International Journal for Parasitology14: 497–501. Infection with M. corti generated significant resistance against a challenge with T. crassiceps introduced either 2 or 6 weeks after primary infection. Challenge infection with T. crassiceps did not influence primary infection with M. corti. Infection with T. crassiceps protected significantly against challenge with M. corti given 2 weeks but not 6 weeks after the primary infection. Challenge infection with M. corti significantly suppressed primary infection with T. crassiceps.     

Acceleration of the growth of tetrathyridial populations of Mesocestoides Corti (Cestoda: Cyclophyllidea) by splenectomy

Acceleration of the growth of tetrathyridial populations of Mesocestoides corti (Cestoda: Cyclophyllidea) by splenectomy. International Journal for Parasitology4, 165–168. Experiments with LDF₁ hybrid mice showed that splenectomy 7 days prior to infection, increases the total biomass of tetrathyridial populations of Mesocestoides corti in hosts of both sexes.This increase is accompanied by a decrease in the size of individuals, and by an increase in the percentage of two-suckered and acephalic forms. Splenectomy thus accelerates both the growth of the biomass of populations and the asexual multiplication of the tetrathyridia.     

Histopathological changes in livers of mice infected with tetrathyridia of Mesocestoides corti and exposed to different environmental temperatures

Novak M. 1982. Histopathological changes in livers of mice infected with tetrathyridia of Mesocestoides corti and exposed to different environmental temperatures. International Journal for Parasitology12: 41–45. Observations on the histopathology of the liver of mice infected with Mesocestoides corti and kept for 20 days p.i. (post-infection) at low (5 ± 1°C), room (21 ± 1°C) or high (35 ± 1°C) temperature revealed that the degree of liver pathology was directly proportional to the intensity of liver infection, which in turn was the result of the temperature effect. The most severe pathological changes occured in the heavily infected organs of mice kept at low temperature, followed by less prominent changes in moderately infected livers of mice kept at room temperature and the smallest changes in lightly infected livers of mice kept at high temperature. The pathological changes in infected and uninfected livers of hosts exposed to different environmental temperatures are described and compared.     

A comparative study of the susceptibility of inbred strains of mice to infection with Mesocestoides corti

White T. R., Thompson R. C. A. and Penhale W. J. 1982. A comparative study of the susceptibility of inbred strains of mice to infection with Mesocestoides corti. International Journal for Parasitology12: 29–33. The susceptibility of 6 strains of inbred mice to infection with Mesocestoides corti was studied following both intraperitoneal and oral inoculation of tetrathyridia. The greatest degree of resistance was seen in C57BL/6 mice and this resistance was independent of route of inoculation. The proliferation of the parasite in C57BL/6 mice was compared with a more susceptible strain (CBA/H) on 7, 14, 21, 35 and 60 days post-infection. Although both strains harboured significantly different parasite burdens during the initial period following infection, these differences were no longer apparent by day 60.     

A nuclear magnetic resonance study of the d-[C6]glucose metabolism of Mesocestoides corti tetrathyridia in the absence and presence of monensin

Novak M. and Blackburn B. J. 1988. A nuclear magnetic resonance study of the d-[¹³C₆]glucose metabolism of Mesocestoides corti tetrathyridia in the absence and presence of monensin. International Journal for Parasitology18: 1029–1033. The effect of monensin on the glucose metabolism of Mesocestoides corti tetrathyridia was studied using ¹H and ¹³C nuclear magnetic resonance (n.m.r.) spectroscopy. Signals due to lactate, succinate, acetate and alanine were identified in the spectra of the excretory products of tetrathyridia fed d-[¹³C₆]glucose in vitro for 120 min. Monensin, at a concentration of 10 μm, inhibited glucose uptake across the brush border of the tetrathyridia, as indicated by a higher level of labelled hexose and lower levels of metabolic end products in ionophore-containing culture medium. The possible action of monensin on the glucose transport mechanism is discussed.     

T-cell dependent collagenous encapsulating response in the mouse liver to Mesocestoides corti (Cestoda)

Pollacco S., Nicholas W.L., Mitchell G.F. and Chaicharn Stewart A. 1978. T-cell dependent collagenous encapsulating response in the mouse liver to Mesocestoides corti (Cestoda). International Journal for Parasitology8: 457–462. Experiments with genetically hypothymic mice show that the tetrathyridial larvae of Mesocestoides corti (Cestoda) multiply much more rapidly in the liver than in normal mice. In the hypothymic mouse, collagen fibres are not laid down and the parasite is not encapsulated as it is in the normal mouse. Encapsulation probably restricts the parasite's multiplication, and it is suggested that the failure to encapsulate the parasite accounts for its more rapid multiplication in the hypothymic mouse. Fibrogenesis and encapsulation is restored to hypothymic mice by transferring syngeneic thymus cells, spleen cells or peritoneal exudate cells. It is concluded that the encapsulation of M. corti is a T-cell dependent process.     

Effects of mebendazole and levamisole on tetrathyridia of Mesocestoides corti in the mouse

Bennet E.-M., Behm C.A. and Bryant C. 1978. Effects of mebendazole and levamisole on tetrathyridia of Mesocestoides corti in the mouse. International Journal for Parasitology8: 463–466. Mebendazole, but not levamisole, administered to mice carrying artificial infections of 50 tetrathyridia of Mesocestoides corti, was effective in killing the parasites. However, simultaneous administration of mebendazole and levamisole was still more effective. Treatment with levamisole before infection had no additional effect.Injection of mice with dead larvae offered some protection against a subsequent challenge with 50 live larvae; however, levamisole did not then improve the anthelmintic efficacy of mebendazole. In mice rendered immunoincompetent by radiation mebendazole was less effective than in non-irradiated controls and levamisole again did not enhance the effect of mebendazole. It is concluded that anthelmintic efficacy of mebendazole depends on its anthelmintic activity supplemented by the host's immune response; and that levamisole stimulates the latter.     

The ultrastructure of the tegument and the digestive caeca of in vitro cultured metacercariae of Fasciola hepatica

Davies C. 1978. The ultrastructure of the tegument and digestive caeca of in vitro cultured metacercariae of Fasciola hepatica. International Journal for Parasitology8: 197–206. The ultrastructure of the tegument and digestive caeca of metacercariae of Fasciola hepatica grown in vitro in two different media is described and compared with the development of these two systems during maturation in vivo. Although the tegument of metacercariae grown in Medium RC showed no development, that of flukes cultured in Medium CS began to produce T-1 and T-2 granules typical of the liver phase of development in vivo. The gastrodermal cells showed some degree of conversion to an adult-like morphology in vitro with the production of typical secretory granules, a limited amount of orientation of the GER and the development of junctional complexes with adjacent parenchyma cells—this was particularly evident in flukes from Medium CS. The growth achieved in each of the culture media is correlated to the degree of development of the tegument and the digestive caeca.     

Gonadectomy and the development of polycephalic tetrathyridia of Mesocestoides corti (Cestoda: Cyclophyllidea) in mice

The number of polycephalic tetrathyridia present in 150-day-old intraperitoneal larval populations of Mesocestoides corti was markedly increased in mice gonadectomized 1 month prior to infection. This effect was more pronounced in male hosts. In both sexes it was inversely correlated with the size of the populations: the smaller the population, the larger was the number of polycephalic tetrathyridia. These forms are probably produced when the separation of daughter individuals lags behind the growth and organogenesis of these peculiar cestodes.     

Sesquiterpene lactones from Vernonia scorpioides and their in vitro cytotoxicity

Fresh leaves of Vernonia scorpioides are widely used in Brazil to treat a variety of skin disorders. Previous in vivo studies with extracts of this species had also demonstrated a high antitumor potential. This paper reports isolation of four sesquiterpene lactones (hirsutinolides and glaucolides), together with diacetylpiptocarphol, 8-acetyl-13-etoxypiptocarphol, luteolin, apigenin, and ethyl caffeate from fresh leaves and flowers of Vernonia scorpioides. The hypothesis that hirsutinolide 3 is formed during extraction was verified theoretically using Density Functional Theory. The effects of isolated compounds on in vitro tumor cells were investigated, as well as their genotoxicity by means of an in vitro comet assay. The results indicate that glaucolide 2 and hirsutinolide 4 are toxic to HeLa cells. These compounds were genotoxic in vitro, a property that appears to be related to the presence of their epoxy groups, which has been a more reliable indication of toxicity than substitution on C-13 or the presence of α,β-unsaturated keto-groups. These results need to be replicated in vivo in order to ascertain their toxicity.     

Mesocestoides corti in the rat: The cellular response in the peritoneal cavity of rats infected with Mesocestoides corti

The tetrathyridium (second larval stage) of Mesocestoides corti elicits an extensive cellular response in the peritoneal cavity of rats which was monitored over a period of 40 days following infection. The total white cell count of female Wistar rats rose in the peritoneal cavity during the first 10 days of infection, then declined slowly. Eosinophilia was characteristic, as it is in most helminth infections. Cellular adherence to the surface of some tetrathyridia was noted. In rats infected with a large number of tetrathyridia, many parasites were found trapped in the mesenteries.     

Wound healing and regenerative response of fragments of the Drosophila wing imaginal disc cultured in vitro

Fragments of imaginal discs of the fruitfly Drosophila undergo growth and pattern regulation when cultured in vivo in adult female hosts for several days prior to metamorphosis in host larvae. Pattern regulation results in either regeneration of excised pattern elements or duplication of elements whose fate map positions lay within the fragment. Initial wound healing along the cut edge of a fragment is thought to be a crucial first step in the process of pattern regulation. We have examined the capacity for wound healing and pattern regulation of fragments (distal halves) of the wing disc cultured in vitro, using the culture system recently reported to support extensive growth and transdetermination of slightly wounded whole imaginal discs in vitro. Our results suggest that disc fragments and whole discs apparently respond differently in the culture system. With disc fragments, wound healing did not occur in vitro. When fragments were first cultured overnight in adult female hosts to allow initial wound healing prior to explantation in vitro, then some volume increase and regeneration of excised portions occurred during 2–3 weeks of culture in vitro. The extent of apparent growth was much less than that reported for whole discs, and the frequency of regeneration in vitro (19%), while highly significant relative to controls not cultured in vitro (0%), was much less than that observed for fragments cultured in vivo (84%). Furthermore the extent of regeneration which occurred in vitro was considerably smaller than that which occurs during regeneration in vivo.     

An efficient synthetic route for preparation of antimycobacterial wollamides and evaluation of their in vitro and in vivo efficacy

A convenient solid phase peptide synthetic (SPPS) route is reported for the preparation of antimycobacterial wollamides. The method is based on on-resin head-to-tail cyclization and is fast, efficient and amenable to automation. The in vitro antimycobacterial activities of the newly synthesized wollamides were evaluated against M. tuberculosis H37Rv (Mtb H37Rv). To assess their drug-likeness, in vitro pharmacokinetic (ADME) profiling was also performed. For wollamides with potent extracellular potency, intracellular activities and in vivo efficacy were determined. The results disclose the potent antimycobacterial (MIC_{Mtb H37Rv}?=?1.1?µM) and suitable drug-like properties of wollamide A (4b). Out of the synthesized wollamides, four compounds (4b–e) exhibited potent intracellular activities against Mtb H37Rv infected human macrophages (IC₅₀?=?0.2–1.3?µM). Results of in vivo blood exposure and efficacy assays for 4d and 4e are discussed.     

Natural products as sources of new fungicides (V): Design and synthesis of acetophenone derivatives against phytopathogenic fungi in vitro and in vivo

A series of acetophenone derivatives (10a–10i, 11, 12a–12g, 13a–13g, 14a–14d and 15a–15l) were designed, synthesized and evaluated for antifungal activities in vitro and in vivo. The antifungal activities of 53 compounds were tested against several plant pathogens, and their structure–activity relationship was summarized. Compounds 10a–10f displayed better antifungal effects than two reference fungicides. Interestingly, the most potent compound 10d exhibited antifungal properties against Cytospora sp., Botrytis cinerea, Magnaporthe grisea, with IC₅₀ values of 6.0–22.6?µg/mL, especially Cytospora sp. (IC₅₀?=?6.0?µg/mL). In the in vivo antifungal assays, 10d displayed the significant protective efficacy of 55.3% to Botrytis cinerea and 73.1% to Cytospora sp. The findings indicated that 10d may act as a potential pesticide lead compound that merits further investigation.     

Localization and associated histopathology of asexually proliferative Mesocestoides corti tetrathyridia (cestoda) infecting mouse mammary glands

Female mice of pregnant random-bred, or unmated BALB/c groups were exposed per os to Mesocestoides corti tetrathyridia and necropsied at various intervals postexposure. The right posterior subcutaneous fat pad with two mammary glands was removed from each mouse, stained, mounted whole and examined microscopically for localization of worms. The left fat pad/gland set was processed, sectioned and stained using standard histological techniques. In pregnant mice, tetrathyridia were localized primarily in the fat pad's posteroventral lobe. Unmated mice had few worms in the mammary glands or associated fat pads. The difference in infection levels between the two host groups may result from mouse strain difference or the pregnant condition of one group.     

Design,synthesis and evaluation of novel N-phenylbutanamide derivatives as KCNQ openers for the treatment of epilepsy

KCNQ (Kv7) has emerged as a validated target for the development of novel anti-epileptic drugs. In this paper, a series of novel N-phenylbutanamide derivatives were designed, synthesized and evaluated as KCNQ openers for the treatment of epilepsy. These compounds were evaluated for their KCNQ opening activity in vitro and in vivo. Several compounds were found to be potent KCNQ openers. Compound 1 with favorable in vitro activity was submitted to evaluation in vivo. Results showed that compound 1 owned significant anti-convulsant activity with no adverse effects. It was also found to posses favorable pharmacokinetic profiles in rat. This research may provide novel potent compounds for the discovery of KCNQ openers in treating epilepsy.     

Variability in azygospore production among Entomophaga maimaiga isolates

This study describes in vitro and in vivo azygospore production by nine isolates of Entomophaga maimaiga, a fungal pathogen of the gypsy moth, Lymantria dispar. The three E. maimaiga isolates that consistently produced azygospores in vitro were also strong producers of azygospores in vivo. However, two additional isolates that were strong azygospore producers in vivo did not produce azygospores in vitro. Isolates that produced azygospores in vitro produced both conidia and azygospores more frequently in vivo than isolates not producing azygospores in vitro. In vitro azygospore production varied over time as well as by isolate. After >2 years of cold storage, while three isolates continued in vitro azygospore production, three isolates no longer produced azygospores in vitro.