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Plasmodium sporozoites injected into the skin by malaria-infected mosquitoes can be effectively targeted by antibodies that block parasite invasion of host hepatocytes and thus prevent the subsequent development of blood stage infections responsible for clinical disease. Malaria subunit vaccines require potent adjuvants, as they lack known pathogen-associated molecular patterns found in attenuated viral or bacterial vaccines that function as Toll-like receptor (TLR) agonists to stimulate dendritic cells and initiate strong adaptive immune responses. A synthetic TLR7 agonist, imiquimod, which is FDA approved for topical treatment of various skin conditions, can function as a potent adjuvant for eliciting T-cell responses to intracellular pathogens and model protein antigens. In the current studies, the topical application of imiquimod at the site of subcutaneously injected Plasmodium falciparum circumsporozoite (CS) peptides elicited strong parasite-specific humoral immunity that protected against challenge with transgenic rodent parasites that express P. falciparum CS repeats. In addition, injection of a simple linear peptide followed by topical imiquimod elicited strong Th1 CD4 ؉ T-cell responses, as well as high antibody titers. The correlation of high anti-repeat antibody titers with resistance to sporozoite challenge in vivo and in vitro supports use of this topical TLR7 agonist adjuvant to elicit protective humoral immunity. The safety, simplicity, and economic advantages of a topical synthetic TLR7 agonist adjuvant also apply to other vaccines requiring high antibody titers, such as malaria asexual or sexual blood stage antigens to prevent red blood cell invasion and block transmission to the mosquito vector, and to vaccines to other extracellular pathogens.The control of the Plasmodium parasite, which causes 300 to 500 million malaria infections and more than 1 million deaths each year, will require a multifaceted approach involving insecticides, chemotherapy, and development of an inexpensive, efficacious malaria vaccine. While significant advances have been made in the use of insecticide-impregnated bed nets and new drug combinations, a licensed malaria vaccine is not yet available. Thus far, the attenuated sporozoite provides the most potent malaria vaccine that can fully protect human volunteers against experimental challenge by the P. falciparuminfected mosquito and thus prevent development of blood stage parasites responsible for clinical disease (13,18,19,43).Significant logistical challenges remain in scale-up production of attenuated parasite vaccines for the 40% of the world's population currently at risk of malaria (28, 44). Sporozoites cannot be grown in vitro and can be obtained only by dissection of salivary glands of mosquitoes that have fed on human blood infected with P. falciparum parasites. Although progress has been made in addressing the regulatory and safety issues related to production of purified P. falciparum sporozoites for vaccines, the requirement for human blood products and t...

1. Specific Ca2+ channel blocker were used to isolate and characterize different components of whole cell Ba2+ current in granule neurons acutely dissociated from guinea pig hippocampal slices. 2. Granule cell Ba2+ current peaked around +5 mV, whether elicited by step or ramp commands, and was completely blocked by 100 microM Cd2+. 3. Saturating doses (> or = 5 microM) of the dihydropyridine antagonist, nimodipine, blocked 39% of the total current, and the inhibition was partially reversible. Nimodipine had a greater effect on current at the end of a depolarizing step, suggesting some voltage dependence of its action. 4. Saturating doses (> or = 3 microM) of omega-conotoxin-GVIA irreversibly blocked 21% of the total current and did not occlude subsequent nimodipine inhibition. 5. High concentrations (> or = 100 nM) of omega-agatoxin-IVA irreversibly blocked another 20% of the total current. The effect of omega-Aga-IVA was quite slow, but saturated after several minutes at 200 nM concentration. A high dose (1 microM) produced rapid effects that were used to quantify the magnitude of block. 6. When applied together, all three blockers inhibited nearly the same amount of total current (77%) as would be expected from the sum of each blocker applied individually (80%), suggesting that the three antagonists blocked different channel types. 7. Quantitatively similar results were obtained for the effect of each blocker alone and in combination on currents elicited by depolarizing ramp commands. Ramp currents blocked by each antagonist (difference ramps) all peaked near the same potential as the initial control ramp, indicating very similar activation properties for the three current components. Difference currents elicited by step commands similarly showed little difference among the three components in their kinetics of inactivation during relatively brief (30 ms) depolarizations. With longer steps, however, the omega-CgTX-GVIA-sensitive component showed some inactivation, whereas the omega-Aga-IVA-sensitive current did not inactivate. 8. The component of current resistant to all three blockers (approximately 23% of total current) was not inhibited by omega-conotoxin-MVIIC, but was half blocked by 50 microM Ni2+. This Ni(2+)-sensitive component showed relatively rapid inactivation and peaked at a somewhat lower potential (-10 mV) than the control current. The resistant current was also inactivated by approximately 50% by holding at -60 compared with -80 mV.(ABSTRACT TRUNCATED AT 400 WORDS)

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SUMMARY1. The isometric properties and related biochemical properties of a developing rat fast muscle (extensor digitorum longus, EDL) have been determined during the 21 days after birth.2. At birth the maximum rate of rise of tension in a tetanus, a measure of speed of contraction, is slow; it begins to increase only after the 5th day and reaches adult values by 21 days. 3. The increase in actomyosin ATPase activity of the rat EDL correlates closely (P < 0.001) with the changes in the maximum rate of rise during the same period of development.4. The relaxation phase of a tetanic response begins to increase in speed promptly after birth, and reaches adult values by the 21st day. The rate of calcium accumulation by the isolated sarcoplasmic reticulum (SR) increases with a similar time course.5. The separate contributions of contraction and relaxation mechanisms to the changes in the more conventional isometric properties of maturing muscles have been analysed.