Possible direct effect of diethylcarbamazine on the infective larvae of Brugia pahangi

1990 ◽  
Vol 64 (4) ◽  
pp. 295-301
Author(s):  
Yasunori Fujimaki ◽  
Masaaki Shimada ◽  
Yoshinori Mitsui ◽  
Eisaku Kimura ◽  
Yoshiki Aoki

ABSTRACTThe direct action of diethylcarbamazine (DEC) on the infective larvae of Brugia pahangi was studied. The larvae were cultured in RPMI 1640 supplemented with foetal bovine serum and antibiotics for 22 days. Most of the larvae remained alive for 8 days, but survival rate of larvae decreased rapidly from day 10 onwards. The larvae did not grow in the culture system. The addition of DEC did not affect the morbidity of the larvae and no difference was observed in the morphological characteristics between the larvae cultured in the presence or absence of DEC.The infective larvae were cultured in vitro for 5 days in the presence or absence of DEC, and inoculated into jirds. The animals were necropsied at intervals, and developing larvae and adult worms were recovered. When the larvae were cultured without DEC and then inoculated subcutaneously into jirds, 29.8% of the inoculum was recovered 3–15 days, and 25% 19–22 weeks, post-inoculation. However, when the larvae were exposed to DEC in vitro and inoculated into jirds, the rate of recovery was reduced to 25% 3–15 days post-inoculation and 2% after 19–22 weeks. When the control larvae cultured in vitro were inoculated intraperitoneally into jirds, 41·3% of inoculum was recovered 3–15 days, and 42·8% 19–22 weeks, post-inoculation. Again the corresponding value for larvae exposed to DEC in vitro was reduced to 19.8% 3–15 days, and 8% 19–22 weeks, post-inoculation. It was observed that the larvae exposed to DEC in vitro were retarded in their development in jirds. These results indicate that DEC has a direct action against the infective larvae of B. pahangi.

1989 ◽  
Vol 122 (1) ◽  
pp. 303-NP ◽  
Author(s):  
B. Fisch ◽  
R. A. Margara ◽  
R. M. L. Winston ◽  
S. G. Hillier

ABSTRACT A primary monolayer cell culture system was developed to investigate human corpus luteum (CL) function in vitro. Steroidogenic cells were isolated by collagenase dispersal and Percoll density-gradient fractionation from CLs enucleated at progressive stages of the luteal phase (tubal surgery patients). 'Pure' granulosa-lutein cells were aspirated from ovulatory follicles at mid-cycle (in-vitro fertilization patients). The steroidogenic capacity (progesterone/20α-dihydroprogesterone biosynthesis and aromatase activity) of isolated luteal cells was assessed in relation to CL development. Basal luteal cell steroidogenesis was maximal at around the expected time of ovulation and declined with CL age during the luteal phase. Conversely, human chorionic gonadotrophin (hCG)-responsive steroidogenesis was initially undetectable but developed as the luteal phase progressed. These results show that luteal cell steroidogenesis becomes increasingly dependent upon gonadotrophic support with CL age. This is evidence that functional luteolysis in human ovaries (1) is pre-programmed to occur at the cellular level, (2) is initiated automatically at the time of ovulation and (3) is reversed at the time of CL 'rescue' in early pregnancy by the direct action of trophoblastic hCG on steroidogenic luteal cells. The culture system described should be of value in further defining the control of human CL form and function at the cellular level. Journal of Endocrinology (1989) 122, 303–311


1976 ◽  
Vol 50 (1) ◽  
pp. 21-28 ◽  
Author(s):  
Rosemary Rogers ◽  
D. A. Denham

AbstractThe effects of levamisole on adults, third stage infective larvae, and microfilariae of Brugia pahangi were studied in in vitro culture and in vivo against developing stages in the vector mosquito and in infected cats. In vitro the drug was effective only at dose levels much higher than can be tolerated by mammals. It was active against the developmental stages of the worm in the vector Aedes aegypti.The drug was strongly microfilaricidal in cats but less effective against adult worms.


2010 ◽  
Vol 25 (6) ◽  
pp. 541-548 ◽  
Author(s):  
Daniela Yukie Sakai Tanikawa ◽  
Nivaldo Alonso ◽  
Marisa Roma Herson ◽  
Monica Beatriz Mathor ◽  
Elia Garcia Caldini ◽  
...  

PURPOSE: In order to circumvent several difficulties that have been met in the routine use of the in vitro keratinocyte cultures using the standard procedure described by Rheinwald and Green, and obtain a more resilient and the least possible immunogeneic skin substitute for a future clinical application, this work studied a new keratinocyte culture system, which envisages the utilization of a fibrin substrate in association with high densities of human keratinocytes. METHODS: Through light and transmission electron microscopy and immunohistochemical assays, long-term proliferative and differentiative characteristics of keratinocytes cultured onto a fibrin gel under immerse and air-liquid interface culture conditions were evaluated. RESULTS: Despite the absence of a dermal substitute, the results demonstrated that the proposed composite was constituted of a transparent and elastic fibrin film covered by a well-attached, multistratified epithelium with morphological characteristics that resemble human epidermis, including the neoformation, albeit incomplete, of the basement membrane. CONCLUSIONS: Increased mechanical resistance due to the presence of an easy handling substrate, the delivery of nonclonfluent keratinocytes as well as the removal of animal-derived cells from the culture system suggest its potential use for future transplantation purposes.


2019 ◽  
Vol 39 (9) ◽  
pp. 1512-1524 ◽  
Author(s):  
Juan A Martín ◽  
Juan Sobrino-Plata ◽  
Begoña Coira ◽  
David Medel ◽  
Carmen Collada ◽  
...  

Abstract The Dutch elm disease (DED) pathogens, Ophiostoma ulmi (Buisman) Nannf. and the more aggressive Ophiostoma novo-ulmi Brasier, have decimated European elm populations in the last 100 years. Today, the number of tolerant elm varieties available on the market is limited, partly due to the long breeding cycles and expensive facilities they require. Developing a low-cost technique to allow early screening of elm tolerance based on simple morphological and/or biochemical traits would considerably boost elm breeding and research. Within this general aim, we developed an in vitro plant culture system to (i) characterize stress responses to O. novo-ulmi-root inoculation in two Ulmus minor Mill. clones of contrasting susceptibility level to DED (termed ‘tolerant’ and ‘susceptible’) and (ii) compare the upward dispersal rate of the pathogen in the two clones. Constitutive xylem anatomy was similar in both clones, indicating that differences in plant responses to the pathogen are not attributable to anatomical factors (e.g., conduit size). Susceptible plantlets suffered a significant delay in apical growth and a decrease in chlorophyll content at 21 days post-inoculation (dpi). The rate of pathogen dispersal from roots to aerial tissues was similar in both clones. However, the tolerant clone showed a marked increase in lipid peroxidation at 1 dpi, while the susceptible clone showed enhanced values of lipid peroxidation during most of the experimental period (1–21 dpi). Despite wide stem colonization by the pathogen, the tolerant clone effectively regulated the oxidative stress levels and showed remarkable resilience to inoculation. These results extend current knowledge on elm defense mechanisms, and the proposed in vitro plant culture system emerges as a promising early screening method for tolerance to improve elm breeding.


Processes ◽  
2020 ◽  
Vol 8 (11) ◽  
pp. 1345
Author(s):  
Sebastian Bunte ◽  
Birgit Keller ◽  
Bussarakam Chuppava ◽  
Josef Kamphues ◽  
Christian Visscher ◽  
...  

Improving the hygienic status of feed ingredients by biotechnological processes as fermentation is of the greatest concern. This preliminary study aimed to investigate whether there are relevant effects of fermented liquid feed (FLF) on the survival of potential pathogens in vitro. The feed (fresh basis) consisted of 50% rye, 30% rapeseed extracted meal, 10% barley and 10% wheat. Glass bottles were filled about 14.1 g water (38 °C) containing the diluted starter culture and feed (8.81 g). Fermentation led to high levels of lactate (5–7% of dry matter), low pH values (<4.0) and low levels of acetic acid (<1% of dry matter) in the FLF. The survival rate of pathogens added, such as Salmonella enterica serovar Typhimurium, Escherichia coli and Clostridium perfringens after 6 h of controlled fermentation, was significantly reduced (<2 log10 CFU/g). The counts of Candida krusei in FLF at 3 h and 6 h post inoculation remained almost unchanged regardless of the incubation time. Even adding sodium-benzoate at a concentration of up to 0.25% in the liquid feed did not reduce the survival of C.krusei during fermentation. Based on this in vitro study, feeding of FLF seems a promising strategy to reduce pathogen transmission but has to be confirmed on natural feeds by pathogens for increasing the hygienic properties.


2011 ◽  
Vol 86 (4) ◽  
pp. 406-409 ◽  
Author(s):  
Y. Mitsui ◽  
M. Miura ◽  
D.A. Bome ◽  
Y. Aoki

AbstractIn vitro chemotactic responses of infective third-stage larvae (L3) of Brugia pahangi to NaCl, Na2HPO4, KCl, K2HPO4, MgCl2 and CaCl2 were assessed. Compared to deionized water as a control, 200 mm NaCl and 100 mm Na2HPO4 significantly attracted L3 (P < 0.01 and P < 0.01), whereas L3 were likely to avoid 200 mm KCl and 100 mm K2HPO4 (P < 0.05 and P < 0.05). L3 showed no significant tendency to avoid or to be attracted to 200 mm CaCl2 and 200 mm MgCl2. Furthermore, NaCl exhibited a significant chemoattractant activity for L3 at a low concentration of 100 mm.


Parasitology ◽  
1979 ◽  
Vol 78 (3) ◽  
pp. 343-354 ◽  
Author(s):  
S. N. Chen ◽  
R. E. Howells

SUMMARYThe 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 mm incubation. Labelling was also located over the surface of the cuticle of the worms, when incubated for a period of 15 to 60 mm in L-[3H]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.


2008 ◽  
Vol 57 (2) ◽  
pp. 179-184 ◽  
Author(s):  
Teruyo Kusaba ◽  
Yasunori Fujimaki ◽  
Albert L. Vincent ◽  
Yoshiki Aoki

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