Cannabis: Exercise performance and sport. A systematic review.

OBJECTIVES: To review the evidence relating to the effect of cannabis on exercise performance. DESIGN: A systematic review of published literature METHODS: Tetrahydrocannabinol (THC) is the principal psychoactive component of cannabis. A search was conducted using PUB med, Medline and Embase searching for cannabis, marijuana, cannabinoids and THC, in sport and exercise; the contents of sports medicine journals for the last 10 years; as well as cross references from journals and a personal collection of reprints. Only English language literature was reviewed and only articles that specified the details of a formal exercise program or protocol. Individuals in rehabilitation or health screening programs involving exercise were included as the study may have identified adverse reactions in the marijuana group. Review articles, opinion pieces, policy statements by sporting bodies and regulatory agencies were excluded. RESULTS: Only 15 published studies have investigated the effects of THC in association with exercise protocols. Of these studies, none showed any improvement in aerobic performance. Exercise induced asthma was shown to be inhibited. In terms of detrimental effects, two studies found that marijuana precipitated angina at a lower work-load (100% of subjects) and strength is probably reduced. Some subjects could not complete an exercise protocol because adverse reactions caused by cannabis. An important finding relevant to drug testing was that aerobic exercise was shown to cause only very small rises (<1ng/mL) in THC concentrations. CONCLUSIONS: THC does not enhance aerobic exercise or strength.

Biochemical and immunological characterization of bacterially expressed and refolded Plasmodium falciparum 42-kilodalton C-terminal merozoite surface protein 1.

Protection against Plasmodium falciparum can be induced by vaccination in animal models with merozoite surface protein 1 (MSP1), which makes this protein an attractive vaccine candidate for malaria. In an attempt to produce a product that is easily scaleable and inexpensive, we expressed the C-terminal 42 kDa of MSP1 (MSP1(42)) in Escherichia coli, refolded the protein to its native form from insoluble inclusion bodies, and tested its ability to elicit antibodies with in vitro and in vivo activities. Biochemical, biophysical, and immunological characterization confirmed that refolded E. coli MSP1(42) was homogeneous and highly immunogenic. In a formulation suitable for human use, rabbit antibodies were raised against refolded E. coli MSP1(42) and tested in vitro in a P. falciparum growth invasion assay. The antibodies inhibited the growth of parasites expressing either homologous or heterologous forms of P. falciparum MSP1(42). However, the inhibitory activity was primarily a consequence of antibodies directed against the C- terminal 19 kDa of MSP1 (MSP1(19)). Vaccination of nonhuman primates with E. coli MSP1(42) in Freund's adjuvant protected six of seven Aotus monkeys from virulent infection with P. falciparum. The protection correlated with antibody-dependent mechanisms. Thus, this new construct, E. coli MSP1(42), is a viable candidate for human vaccine trials.

T cell tolerance to a neo-self antigen expressed by thymic epithelial cells: the soluble form is more effective than the membrane-bound form.

We have previously shown that transgenic (Tg) mice expressing either soluble or membrane-bound hen egg lysozyme (sHEL or mHEL, respectively) under control of the alphaA-crystallin promoter develop tolerance due to thymic expression of minuscule amounts of HEL. To further address the mechanisms by which this tolerance develops, we mated these two lines of Tg mice with the 3A9 line of HEL-specific TCR Tg mice, to produce double-Tg mice. Both lines of double-Tg mice showed deletion of HEL-specific T cells, demonstrated by reduction in numbers of these cells in the thymus and periphery, as well as by reduced proliferative response to HEL in vitro. In addition, the actual deletional process in thymi of the double-Tg mice was visualized in situ by the TUNEL assay and measured by binding of Annexin V. Notably, the apoptosis localized mainly in the thymic medulla, in line with the finding that the populations showing deletion and increased Annexin V binding consisted mainly of single- and double-positive thymocytes. Interestingly, the thymic deletional effect of sHEL was superior to that of mHEL in contrast to the opposite differential tolerogenic effects of these HEL forms on B cells specific to this Ag. Analysis of bone marrow chimeras indicates that both forms of HEL are produced by irradiation-resistant thymic stromal cells and the data suggest that sHEL is more effective in deleting 3A9 T cells due mainly to its higher accessibility to cross-presentation by dendritic APC.

In vitro studies with recombinant Plasmodium falciparum apical membrane antigen 1 (AMA1): production and activity of an AMA1 vaccine and generation of a multiallelic response.

Apical membrane antigen 1 (AMA1) is regarded as a leading malaria blood-stage vaccine candidate. While the overall structure of AMA1 is conserved in Plasmodium spp., numerous AMA1 allelic variants of P. falciparum have been described. The effect of AMA1 allelic diversity on the ability of a recombinant AMA1 vaccine to protect against human infection by different P. falciparum strains is unknown. We characterize two allelic forms of AMA1 that were both produced in Pichia pastoris at a sufficient economy of scale to be usable for clinical vaccine studies. Both proteins were used to immunize rabbits, singly and in combination, in order to evaluate their immunogenicity and the ability of elicited antibodies to block the growth of different P. falciparum clones. Both antigens, when used alone, elicited high homologous anti-AMA1 titers, with reduced strain cross-reactivity. Similarly, sera from rabbits immunized with a single antigen were capable of blocking the growth of homologous parasite strains at levels theoretically sufficient to clear parasite infections. However, heterologous inhibition was significantly reduced, providing experimental evidence that AMA1 allelic diversity is a result of immune pressure. Encouragingly, rabbits immunized with a combination of both antigens exhibited titers and levels of parasite inhibition as good as those of the single-antigen-immunized rabbits for each of the homologous parasite lines, and consequently exhibited a broadening of allelic diversity coverage.

Vaccination of monkeys with recombinant Plasmodium falciparum apical membrane antigen 1 confers protection against blood-stage malaria.

A major challenge facing malaria vaccine development programs is identifying efficacious combinations of antigens. To date, merozoite surface protein 1 (MSP1) is regarded as the leading asexual vaccine candidate. Apical membrane antigen 1 (AMA1) has been identified as another leading candidate for an asexual malaria vaccine, but without any direct in vivo evidence that a recombinant form of Plasmodium falciparum AMA1 would have efficacy. We evaluated the efficacy of a form of P. falciparum AMA1, produced in Pichia pastoris, by vaccinating Aotus vociferans monkeys and then challenging them with P. falciparum parasites. Significant protection from this otherwise lethal challenge with P. falciparum was observed. Five of six animals had delayed patency; two of these remained subpatent for the course of the infection, and two controlled parasite growth at <0.75% of red blood cells parasitized. The protection induced by AMA1 was superior to that obtained with a form of MSP1 used in the same trial. The protection induced by a combination vaccine of AMA1 and MSP1 was not superior to the protection obtained with AMA1 alone, although the immunity generated appeared to operate against both vaccine components.

Merozoite surface protein 3 and protection against malaria in Aotus nancymai monkeys.

A blood-stage vaccine based on Plasmodium falciparum merozoite surface protein 3 (MSP3) was tested for efficacy in a primate model. Aotus nancymai monkeys were vaccinated with yeast-expressed MSP3 before a lethal challenge with Plasmodium falciparum parasites. Five of 7 control monkeys had acute infections and required treatment to control parasitemia. Only 1 of 7 monkeys vaccinated with MSP3 required this treatment. The efficacy of the MSP3 vaccination appeared to be comparable to that of MSP1(42), a leading asexual vaccine candidate, in response to which 2 monkeys experienced acute infections. In the MSP3-vaccinated group, protection correlated with prechallenge titers of antibody to MSP3. In the MSP1 and control groups, protection correlated with antibody to MSP3 raised by challenge infection.

A recombinant vaccine expressed in the milk of transgenic mice protects Aotus monkeys from a lethal challenge with Plasmodium falciparum.

Two strains of transgenic mice have been generated that secrete into their milk a malaria vaccine candidate, the 42-kDa C-terminal portion of Plasmodium falciparum merozoite surface protein 1 (MSP1(42)). One strain secretes an MSP1(42) with an amino acid sequence homologous to that of the FVO parasite line, the other an MSP1(42) where two putative N-linked glycosylation sites in the FVO sequence have been removed. Both forms of MSP1(42) were purified from whole milk to greater than 91% homogeneity at high yields. Both proteins are recognized by a panel of monoclonal antibodies and have identical N termini, but are clearly distinguishable by some biochemical properties. These two antigens were each emulsified with Freund's adjuvant and used to vaccinate Aotus nancymai monkeys, before challenge with the homologous P. falciparum FVO parasite line. Vaccination with a positive control molecule, a glycosylated form of MSP1(42) produced in the baculovirus expression system, successfully protected five of six monkeys. By contrast, vaccination with the glycosylated version of milk-derived MSP1(42) conferred no protection compared with an adjuvant control. Vaccination with the nonglycosylated, milk-derived MSP1(42) successfully protected the monkeys, with 4/5 animals able to control an otherwise lethal infection with P. falciparum compared with 1/7 control animals. Analysis of the different vaccines used suggested that the differing nature of the glycosylation patterns may have played a critical role in determining efficacy. This study demonstrates the potential for producing efficacious malarial vaccines in transgenic animals.

The cochlear nuclei in Colubrid and boid snakes: A qualitative and quantitative study.

The cochlear nuclear complex was investigated in snakes of the advanced family Colubridae and the primitive family Boidae. This study was undertaken in an attempt to correlate the elaboration of the cochlear nuclei with behavior and phylogeny and to elucidate the relative effects of these factors on the evolution of the cochlear nuclear complex. Fifty-five brains, of 14 colubrid species and three boid species, were examined to collect data on neuron diameter, neuron population, nuclear volume, and neuronal density of the cochlear nuclear complex and of its component nuclei (nucleus angularis and nucleus magnocellularis). Intraspecific and interspecific comparisons of the data were performed by nested analysis of variance. The species were grouped by cluster analysis and ranked on the basis of the morphometric parameters. Interspecific comparisons indicate that the elaboration of the cochlear nuclei is related, first, to prey preference and, second, to habitat preference. The most elaborate cochlear nuclei occur in species with a preference for vertebrate prey. Burrowing species that prey on vertebrates exhibit the highest degree of elaboration of the cochlear nuclei. In some burrowing species, the nucleus magnocellularis is differentiated into medial and lateral subdivisions. The primitive boid snakes show greater elaboration of the cochlear nuclei than do most of the advanced colubrid snakes. The elaboration of the cochlear nuclear complex in snakes seems to reflect the influence of both behavior and phylogeny. Further investigation of primitive snakes of varied behaviors is needed to establish more clearly the influence of phylogeny on the evolution of the cochlear nuclear complex.

The cytoarchitecture of the torus semicircularis in the red-eared turtle.

The cytoarchitecture and neuronal morphology of the torus semicircularis in the red-eared turtle, Chrysemys scripta elegans, were examined in Nissl-stained and Golgi-impregnated material. The torus semicircularis begins in the caudodorsal mesencephalon and extends rostrally and laterally to end ventrally to the tectal ventricle. The torus semicircularis consists of a central nucleus and a laminar nucleus, which is interposed between the central nucleus and the ventricle. The central nucleus can be divided into two regions, a small, large-celled area, located dorsally, and a larger area of small spherical (6-17 µm), large spherical (18-25 µm), triangular (15-27 µm) and fusiform (10-26 µm) neurons. The small spherical cells have two dendritic patterns: "radiate" and "single." The radiate pattern has a dorsoventral orientation, several secondary branches and few dendritic spines. These cells are usually located in the center of the central nucleus. The single pattern is oriented mediolaterally. This cell type is most often observed at the periphery of the central nucleus. These neurons have few secondary branches and dendritic spines. The large spherical neurons display two dendritic orientations: dorsoventral and mediolateral. All dendritic trees have numerous secondary branches and few dendritic spines. The triangular neurons exhibit primary dendrites projecting from the corners of the somata and have few secondary branches and dendritic spines. The fusiform neurons have a majority of their dendrites oriented mediolaterally, few secondary branches and a small number of dendritic spines. The laminar nucleus consists of several layers and three cell types: ovoid (9-15 µm), triangular (20-40 µm), and fusiform (20-40 µm). All neurons have few secondary dendritic branches and few dendritic spines. The dendrites of many neurons course perpendicularly to the long axis of the brainstem and encapsulate the central nucleus. Some ovoid and fusiform neurons display dendrites that enter the central nucleus.

The torus semicircularis in a gekkonid lizard.

The cytoarchitecture and neuromorphology of the torus semicircularis in the tokay gecko, Gekko gecko, were examined in Nissl-stained, fiber-stained, and Golgi-impregnated tissues. From a superficial position, the torus semicircularis extends rostrally under the caudal half of the optic tectum. Caudally, the two tori abut upon one another; rostrally, they diverge. The torus semicircularis consists of central, laminar, and superficial nuclei. The central nucleus consists of fusiform, spherical and triangular neurons. Their dendrites are highly branched, with numerous dendritic spines, and are oriented mediolaterally, dorsoventrally, and rostrocaudally. Fusiform and spherical neurons display two dendritic patterns: "single axis," ramifying in one axis, and "dual axis," exhibiting higher-order branches perpendicular to the primary dendrites. Triangular neurons exhibit a "radiate" dendritic pattern. In the rostral half of the torus semicircularis, the laminar nucleus caps the central nucleus. The laminar nucleus encircles the central nucleus in the caudal torus semicircularis. The neurons of the laminar nucleus have dendritic arrays oriented parallel to the border of the central nucleus. These dendrites exhibit a paucity of dendritic spines and higher-order branches. Fusiform and spherical neurons exhibit "single axis" and "dual axis" dendritic patterns. Triangular neurons display "radiate" patterns. The caudal superficial nucleus lies dorsal and dorsolateral to the central nucleus. The superficial nucleus is sparsely populated by small fusiform and spherical neurons with moderately branched dendrites and moderate numbers of dendritic spines. These neurons display "single axis" (fusiform neurons) as well as "dual axis" and "radiate" (spherical neurons) dendritic patterns. They are oriented either parallel to or perpendicular to the boundary of the laminar nucleus.