A brief illustrated guide to the ultrastructure of Plasmodium falciparum asexual blood stages

Interpretation of the new information arising from the Plasmodium falciparum Genome Project requires a good working knowledge of the ultrastructure of the parasite; however many aspects of the morphology of this species remain obscure. Lawrence Bannister, John Hopkins and colleagues here give an illustrated overview of the three-dimensional (3-D) organization of the merozoite, ring, trophozoite and schizont stages of the parasite, based on available data that include 3-D reconstruc-tion from serial electron microscope sections. The review describes the chief organelles present in these stages, emphasizing the continuity of structure in addition to specialized, stage-specific features developed during the asexual erythrocytic cycle.     

Mitochondrial malic enzyme activity is much higher in mitochondria from cortical synaptic terminals compared with mitochondria from primary cultures of cortical neurons or cerebellar granule cells

Most of the malic enzyme activity in the brain is found in the mitochondria. This isozyme may have a key role in the pyruvate recycling pathway which utilizes dicarboxylic acids and substrates such as glutamine to provide pyruvate to maintain TCA cycle activity when glucose and lactate are low. In the present study we determined the activity and kinetics of malic enzyme in two subfractions of mitochondria isolated from cortical synaptic terminals, as well as the activity and kinetics in mitochondria isolated from primary cultures of cortical neurons and cerebellar granule cells. The synaptic mitochondrial fractions had very high mitochondrial malic enzyme (mME) activity with a Km and a Vmax of 0.37 mM and 32.6 nmol/min/mg protein and 0.29 mM and 22.4 nmol/min mg protein, for the SM2 and SM1 fractions, respectively. The Km and Vmax for malic enzyme activity in mitochondria isolated from cortical neurons was 0.10 mM and 1.4 nmol/min/mg protein and from cerebellar granule cells was 0.16 mM and 5.2 nmol/min/mg protein. These data show that mME activity is highly enriched in cortical synaptic mitochondria compared to mitochondria from cultured cortical neurons. The activity of mME in cerebellar granule cells is of the same magnitude as astrocyte mitochondria. The extremely high activity of mME in synaptic mitochondria is consistent with a role for mME in the pyruvate recycling pathway, and a function in maintaining the intramitochondrial reduced glutathione in synaptic terminals.     

Polymorphisms in genes involved in folate metabolism as maternal risk factors for Down syndrome

Down syndrome is a complex genetic and metabolic disorder attributed to the presence of three copies of chromosome 21. The extra chromosome derives from the mother in 93% of cases and is due to abnormal chromosome segregation during meiosis (nondisjunction). Except for advanced age at conception, maternal risk factors for meiotic nondisjunction are not well established. A recent preliminary study suggested that abnormal folate metabolism and the 677C-->T polymorphism in the methylenetetrahydrofolate reductase (MTHFR) gene may be maternal risk factors for Down syndrome. The present study was undertaken with a larger sample size to determine whether the MTHFR 677C-->T polymorphism was associated with increased risk of having a child with Down syndrome. Methionine synthase reductase (MTRR) is another enzyme essential for normal folate metabolism. A common polymorphism in this gene was recently associated with increased risk of neural tube defects and might also contribute to increased risk for Down syndrome. The frequencies of the MTHFR 677C-->T and MTRR 66A-->G mutations were evaluated in DNA samples from 157 mothers of children with Down syndrome and 144 control mothers. Odds ratios were calculated for each genotype separately and for potential gene-gene interactions. The results are consistent with the preliminary observation that the MTHFR 677C-->T polymorphism is more prevalent among mothers of children with Down syndrome than among control mothers, with an odds ratio of 1.91 (95% confidence interval [CI] 1.19-3.05). In addition, the homozygous MTRR 66A-->G polymorphism was independently associated with a 2. 57-fold increase in estimated risk (95% CI 1.33-4.99). The combined presence of both polymorphisms was associated with a greater risk of Down syndrome than was the presence of either alone, with an odds ratio of 4.08 (95% CI 1.94-8.56). The two polymorphisms appear to act without a multiplicative interaction.     

Outcomes of breeding soundness evaluation of 2898 yearling bulls subjected to different classification systems

This study is a retrospective analysis comparing data on yearling bull breeding soundness evaluation (BSE) subjected to 3 different classification systems: the Society for Theriogenology (SFT) 1983 and 1993 systems, and the Western Canadian Association of Bovine Practitioners (WC) 1993 system. Data were collected at 5 performance bull-test stations located in South Carolina and Tennessee from 1986 through 1996. Yearling bulls (n=2898) that were 10 to 20 mo of age were used in the analysis. To simplify analysis, bulls were determined to be either satisfactory or unsatisfactory potential breeders (including those classified as questionable, deferred or unsatisfactory). Data were analyzed 1) within location where a location was treated as an individual experiment, 2) with breeds and locations collapsed, and 3) within age-group where each age-group was treated as an individual experiment. An ANOVA was performed using GLM procedures of SAS, and this model was used to generate least square means for the proportion of bulls classified as satisfactory and the 5 possible unsatisfactory outcomes due to inadequacies in scrotal circumference, spermatozoal morphology, spermatozoal motility, a combination of inadequate spermatozoal morphology and motility, or physical abnormalities. Inadequate scrotal circumference or physical abnormality did not affect differences for BSE outcomes among systems. Using the SFT93 system, bulls failed at a higher rate due to inadequate spermatozoal morphology (P < 0.05) than when subjected to the SFT83 system. In the WC93 system, a higher percentage of bulls failed due to inadequate spermatozoal motility and to a combination of inadequate spermatozoal morphology and motility than in the other 2 systems (P < 0.05). The proportion of bulls in this data failing under the WC93 system appears to be the result of overestimation.     

Human homologue of S. pombe Rad9 interacts with BCL-2/BCL-xL and promotes apoptosis

DNA damage induces apoptosis through a signalling pathway that can be suppressed by the BCL-2 protein, but the mechanism by which DNA damage does this is unknown. Here, using yeast two-hybrid and co-immunoprecipitation studies, we show that RAD9, a human protein involved in the control of a cell-cycle checkpoint, interacts with the anti-apoptotic Bcl-2-family proteins BCL-2 and BCL-x L, but not with the pro-apoptotic BAX and BAD. When overexpressed in mammalian cells, RAD9 induces apoptosis that can be blocked by BCL-2 or BCL-x L. Conversely, antisense RAD9 RNA suppresses cell death induced by methyl methanesulphonate. These findings indicate that RAD9 may have a new role in regulating apoptosis after DNA damage, in addition to its previously described checkpoint-control and other radioresistance-promoting functions.     

Endocrine profiles of dairy cows following experimentally induced clinical mastitis during early lactation

Concentrations of LH, cortisol, estradiol-17beta (E(2)), prolactin and 13,14-dihydro-15-keto-prostaglandin F(2alpha) (PGFM) were determined in cows with experimentally induced clinical mastitis during early lactation. Cows free of intramammary infection (IMI) and in the luteal phase of the estrous cycle were balanced by lactation number and days in milk and assigned to either control (n=5) or treatment (n=5) groups. Treated cows were infected experimentally (day 0), in two mammary quarters, with Streptococcus uberis and developed clinical mastitis within 60 h after inoculation as evidenced by increased mastitis scores, elevated rectal temperatures, mammary swelling and isolation of S. uberis pathogen. Four days following bacterial challenge, blood samples were collected every 20 min for 8 h for determination of PGFM and LH following administration of oxytocin and GnRH, respectively. Blood samples were also collected on days 0, 4 and 7 of the experiment to determine concentrations of E(2), prolactin and cortisol. Four days after bacterial challenge, concentrations of cortisol were higher (P=0.04) in experimentally infected cows than controls. Experimentally challenged cows had increased (P=0.02) concentrations of cortisol on days 4 and 7 compared with day 0. Control cows had no significant increase in blood cortisol during the experimental period. Baseline concentrations of PGFM did not differ between groups; however, peak concentrations of PGFM following oxytocin challenge were elevated (P=0.006) in cows with clinical mastitis compared with control animals. Prolactin, E(2) and LH did not differ between cows with clinical mastitis or controls. Experimentally induced mastitis during early lactation elevated concentrations of cortisol during the luteal phase of the estrous cycle. Furthermore, mastitic cows demonstrated an increased PGFM response following oxytocin administration. Altered reproductive efficiency in cows with clinical mastitis caused by Gram-positive pathogens may be the result of increased uterine sensitivity to prostaglandin F(2alpha) (PGF(2alpha)).     

Sequence-dependent dynamics in duplex DNA

The submicrosecond bending dynamics of duplex DNA were measured at a single site, using a site-specific electron paramagnetic resonance active spin probe. The observed dynamics are interpreted in terms of the mean squared amplitude of bending relative to the end-to-end vector defined by the weakly bending rod model. The bending dynamics monitored at the single site varied when the length and position of a repeated AT sequence, distant from the spin probe, were changed. As the distance between the probe and the AT sequence was increased, the mean squared amplitude of bending seen by the probe due to that sequence decreased. A model for the sequence-dependent internal flexural motion of duplex DNA, which casts the mean squared bending amplitudes in terms of sequence-dependent bending parameters, has been developed. The best fit of the data to the model occurs when the (AT)(n) basepairs are assumed to be 20% more flexible than the average of the basepairs within the control sequence. These findings provide a quantitative basis for interpreting the kinetics of biological processes that depend on duplex DNA flexibility, such as protein recognition and chromatin packaging.     

Adjunctive treatment of congenital pigmented nevi with phenol chemical peel

The purpose of this study was a retrospective evaluation of the treatment of congenital pigmented nevi using the phenol chemical peel technique. Patients were treated with standard Baker formula in the operating room under general anesthesia or intravenous sedation with continuous electrocardiogram monitoring. A total of 20 patients were reviewed (13 girls and 7 boys, mean age 3.8 years). Eight patients had nevi located on the face, five patients had trunk lesions, and three patients had lesions on the thighs. Two patients had nevi located on both the face and the trunk, and two patients had involvement of the face, trunk, and thigh. Three of the above patients had the classic "bathing trunk" distribution of the nevi. A test area was peeled in four patients, and in five patients preoperative biopsies were performed to rule out malignancy before initiation of therapy. An average of 2.6 treatments were performed per patient. Two patients had adjunctive dermabrasion to increase the depth of peel and to contour surface irregularities. The length of follow-up ranged from 6 to 84 months with a mean of 28 months. Healing of the wounds occurred within 2 to 3 weeks postoperatively. Seventy-five percent of patients had satisfactory cosmetic improvement in the appearance of the lesions following treatment. Four patients had recurrence of the pigmentation after an initial lightening response, three of whom had their nevi subsequently excised. There was no incidence of hypertrophic scarring or cardiac and/or renal complications. There was one death from complications of leptomeningeal melanocytosis. Chemical peeling of congenital pigmented nevi is an acceptable alternative method of therapy for those lesions that are too large for excision and primary closure or for lesions in which excision would result in unacceptable scars in areas such as the face.