Identification of Apoptotic Bodies in Equine Semen

Apoptosis in the testis is required to ensure an efficient spermatogenesis. However, sometimes, defective germ cells that are marked for elimination during this process escape elimination in the testes, giving rise to ejaculates with increased percentages of abnormal and apoptotic spermatozoa and a high percentage of apoptotic bodies. Apoptosis markers in the ejaculate have been associated with low fertility, either in animals or humans. Therefore, the goal of this study was to investigate whether fresh equine semen contains apoptotic bodies [initially named Merocyanine 540 (M540) bodies] and to study the relationship between the quantity of these bodies and cell concentration, the volume of ejaculate, viability and motility. Moreover, we also studied whether the presence apoptotic bodies in fresh semen was related to the resistance of the stallion spermatozoa to being incubated at 37°C or being frozen and thawed. Fresh equine semen was stained with fluorescent dyes such as M540 and Annexin‐V. Active Caspase 3 was studied in fresh semen through Western blotting and immunofluorescence with a specific antibody. Sperm kinematics was assessed in fresh, incubated and thawed samples using computer‐assisted semen analysis, and viability was evaluated with the LIVE/DEAD Sperm Viability Kit. Overall, our results demonstrate for the first time the presence of apoptotic bodies in equine semen. The quantity of apoptotic bodies was highly variable among stallions and was positively correlated with Caspase 3 activity in fresh samples and negatively correlated with the viability and motility of stallion spermatozoa after the cryopreservation process.     

Oncogenic CARD11 mutations in human diffuse large B cell lymphoma

Diffuse large B cell lymphoma (DLBCL) is the most common form of non-Hodgkin's lymphoma. In the least curable (ABC) subtype of DLBCL, survival of the malignant cells is dependent on constitutive activation of the nuclear factor-kappaB (NF-kappaB) signaling pathway. In normal B cells, antigen receptor-induced NF-kappaB activation requires CARD11, a cytoplasmic scaffolding protein. To determine whether CARD11 contributes to tumorigenesis, we sequenced the CARD11 gene in human DLBCL tumors. We detected missense mutations in 7 of 73 ABC DLBCL biopsies (9.6%), all within exons encoding the coiled-coil domain. Experimental introduction of CARD11 coiled-coil domain mutants into lymphoma cell lines resulted in constitutive NF-kappaB activation and enhanced NF-kappaB activity upon antigen receptor stimulation. These results demonstrate that CARD11 is a bona fide oncogenein DLBCL, providing a genetic rationale for the development of pharmacological inhibitors of the CARD11 pathway for DLBCL therapy.     

The extreme Kuiper Belt binary 2001 QW322

The study of binary Kuiper Belt objects helps to probe the dynamic conditions present during planet formation in the solar system. We report on the mutual-orbit determination of 2001 QW322, a Kuiper Belt binary with a very large separation whose properties challenge binary-formation and -evolution theories. Six years of tracking indicate that the binary's mutual-orbit period is approximately 25 to 30 years, that the orbit pole is retrograde and inclined 50 degrees to 62 degrees from the ecliptic plane, and, most surprisingly, that the mutual orbital eccentricity is <0.4. The semimajor axis of 105,000 to 135,000 kilometers is 10 times that of other near-equal-mass binaries. Because this weakly bound binary is prone to orbital disruption by interlopers, its lifetime in its present state is probably less than 1 billion years.     

High frequency of hypermutable Pseudomonas aeruginosa in cystic fibrosis lung infection

The lungs of cystic fibrosis (CF) patients are chronically infected for years by one or a few lineages of Pseudomonas aeruginosa. These bacterial populations adapt to the highly compartmentalized and anatomically deteriorating lung environment of CF patients, as well as to the challenges of the immune defenses and antibiotic therapy. These selective conditions are precisely those that recent theoretical studies predict for the evolution of mechanisms that augment the rate of variation. Determination of spontaneous mutation rates in 128 P. aeruginosa isolates from 30 CF patients revealed that 36% of the patients were colonized by a hypermutable (mutator) strain that persisted for years in most patients. Mutator strains were not found in 75 non-CF patients acutely infected with P. aeruginosa. This investigation also reveals a link between high mutation rates in vivo and the evolution of antibiotic resistance.     

Genetic and statistical models for estimating genetic parameters of maize seedling resistance to Fusarium graminearum Schwabe root rot

Root lodging is an important problem in corn fields. Fungi recovered from roots include seedling blight and stalk rot pathogens. The objective of this work was to study the inheritance of maize seedling resistance to pathogens causing maize lodging. The Fusarium graminearum strain, 241 Fr1, was isolated from maize lodged plants and identified as the most pathogenic isolate for root rotting. Nine inbred lines of maize and their diallel F₁ crosses plus control genotypes were studied. Seedlings were inoculated at the stage of four-leaves. Disease severity was measured as percentage of the root rotted area. Highly significant differences between inoculated and non-inoculated genotypes were found. Four genetic models and two statistical approaches—the mixed model for the best linear unbiased prediction (BLUP) and the general linear model (GLM)—were used for the analysis. Favorable heterosis of resistance of hybrids over inbreds was the most important effect detected. The general combining ability (GCA) effects were significant for all genetic models and statistical methods studied, and a good agreement existed among the GCA estimates by the different methods. The type of gene action, either additive or dominance, showed a large variation among the parental inbreds and hybrids. Selection of additive effects based exclusively on inbred lines is not sufficient to confer resistance to hybrids, additional selection should be practiced on hybrids to look for favorable dominance effects. This revised version was published online in July 2006 with corrections to the Cover Date.     

The use of hand-raised psittacines for reintroduction: a case study of scarlet macaws (Ara macao) in Peru and Costa Rica

This study reports on three scarlet macaw (Ara macao) reintroduction projects using hand-raised birds in Peru and Costa Rica. The habitats at the release sites ranged from pristine tropical forest to forest fragments in an agricultural matrix. The combined first-year survival was 74% and the annual post first-year survival was 96%. Survival rates were very high despite a wide range in predator communities. Number of birds released explained 70% of the variation in survival with birds from larger releases having higher survival rates. Behavioral evidence suggests that birds established at the site facilitated survival of later releases. Breeding attempts were recorded at all three sites and hand-raised birds with wild mates successfully fledged young in Peru. Supplemental feeding post-release played an important role in keeping the birds near the release site and facilitating social interactions. This work shows that properly socialized hand-raised macaws can survive and breed in the wild but that ex-pets are not good release candidates.     

A comparative study of apple and pear tree water consumption measured with two weighing lysimeters

A five-year experiment (2002–2006) was conducted to determine apple (cv ‘Golden Smoothee’) and pear (cv ‘Conference’) crop coefficients (Kc) using two large weighing lysimeters. Daily reference evapotranspiration (ETo) and crop evapotranspiration (ETc) were evaluated. Midday canopy light interception of both crops planted in hedgerows, 4 × 1.6 m, was determined on a weekly basis from bud-break until leaf fall from year 2002 (fourth after planting) to year 2006 of both plantations. Relationships between canopy light interception and calculated Kc (ETc/ETo) were evaluated from bud-break until harvest. There were differences in Kc values between apple and pear trees. When daily Kc values from bud-break until harvest were adjusted to hyperbolic functions each year, adjusted curves for pear trees were very similar regardless of year (maximum Kc around 1.0). In apple trees, the maximum values of Kc increased over time, from 0.49 in 2002 to 1.04 in 2006. Midday light interception in both apple and pear trees increased during the 5 years of experiment from 29.0 to 45.6% in apples and from 27.5 to 41.6% in pears in midsummer. Although there was a significant positive correlation between midday canopy light interception and Kc in apple and pear trees, in different times within a specific year, these relationships were different between crops. While the apple data fitted into the same equation regardless of the year, different equations were needed to fit the pear data in different years. This discrepancy may have been related to differences in the canopy properties between apple and pear trees. Pear canopies had higher porosity than apple canopies and thus improved light penetration. Apple trees were more vigorous and produced taller and denser canopies. Pear Kc values were greatly influenced by the evaporative demands of different years and consequently differences in midday canopy light interception did not adequately reflect the differences in Kc across the two species.