Comparison of different methods for diagnosis of bovine tuberculosis from tuberculin- or interferon-gamma-reacting cattle in Spain

Of 1479 cattle from herds in Northwestern Spain previously diagnosed as tuberculosis (TB) positive, 218 animals which gave a positive tuberculin or interferon-gamma reaction were examined at the slaughterhouse. Medial retropharyngeal and caudal mediastinal lymph nodes, and any tissues containing lesions suspected to be tuberculous, were removed and submitted to the laboratory. Three techniques for diagnosis of TB were used: post mortem examination (PME), smear staining by means of auramine O method (AOM), and culture isolation in Coletsos and Lowenstein-Jensen media followed by confirmation of M. tuberculosis complex organisms using PCR (CIM-PCR). Only 123 (29.9%) of the 412 samples collected showed typical tuberculous lesions. Confirmed M. tuberculosis complex organisms were isolated in 144 cases, 114 of which were from tissues showing lesions (success rate of 92.8%). Smears were found positive in 113 cases, 96 of which came from lesions suspected to be tuberculous (success rate of 78.0%). The sensitivities of CIM-PCR compared with those of PME and AOM were 92.7% and 85.7%, respectively. Statistical analysis revealed that PME and AOM are good indicators of the presence of M. tuberculosis complex organisms in tuberculin- or interferon-gamma reacting cattle.     

Nucleotide sequence of the D-loop region of the sperm whale (Physeter macrocephalus) mitochondrial genome

We have amplified, by the polymerase chain reaction, and have sequenced the D-loop region of the mitochondrial DNA from the sperm whale (Physeter macrocephalus). The sperm whale D-loop was aligned with D- loop sequences from four other cetaceans (Commerson's dolphin, orca, fin whale, and minke whale) and an out-group (cow). This alignment showed the sperm whale sequence to be larger than that of other cetaceans. In addition, some sequence blocks were highly conserved among all six species, suggesting roles in the functioning of mitochondrial DNA. Other blocks that were previously reported to be well conserved among cetaceans showed little sequence conservation with the sperm whale D-loop, which argues against the functional importance of these sequence blocks in cetaceans.      

Characterization of the transferrin-iron uptake system in Neisseria meningitidis

Abstract The transferrin-iron uptake system of six Neisseria meningitidis strains was characterized using ¹²⁵I-transferrin in receptor assays and ⁵⁵Fe-loaded transferrin in uptake assays. Receptors for transferrin varied among the strains both in number (from 700 to 4700 receptors per cell) and in their affinity constants for the protein ( K _a ranged from 0.7×10⁷ to 4.0×10⁷ 1 mol⁻¹). Neither receptor numbers nor affinity constants were significantly different in carrier and invasive strains, although the K_a seem to be somewhat higher in the latter. Iron uptake from transferrin was also variable among the strains, but showed the same lack of correlation with their origin.     

Recent evolutionary history of American Onchocerca volvulus, based on analysis of a tandemly repeated DNA sequence family

Polymerase chain reaction (PCR) products were characterized for a repeated sequence family (designated "O-150") of the human filarial parasite Onchocerca volvulus. In phylogenetic inferences, the O-150 sequences clustered into closely related groups, suggesting that concerted evolution maintains sequence homology in this family. Using a novel mathematical model based on a nested application of an analysis of variance, we demonstrated that African rainforest and savannah strain parasite populations are significantly different. In contrast, parasites collected in the New World are indistinguishable from African savannah strains of O. volvulus. This finding supports the hypothesis that onchocerciasis was recently introduced into the New World, possibly as a result of the slave trade.      

Influence of the nuclear and extranuclear substitution on the singlet molecular oxygen [O 2(1Δg)]-mediated photooxidation of tyrosine derivatives: A kinetic study

Summary The effect of the substitution pattern on the kinetics of the Type II (O₂(¹_g)-mediated) dye-sensitized photooxidation of a series of nine tyrosine derivatives was investigated. Overall (k_t) and reactive (k_r) rate constants for the interaction of the excited oxygen species with the amino acid derivatives were determined. A parallel study on solvent and pH effects was carried out.The presence of different substituents in nuclear positions or in the amino acid side chain greatly affect the photooxidation rates.An upper limit for photooxidation quantum yield, calculated from the kinetic data, varies from 0.03 to 0.25, being the higher for halogenated tyrosines and the lower for esterified tyrosines and for the nitro-derivative.The variation of solvent polarity and pH of the reaction medium confirm that the presence of the ionized phenolate group in tyrosine, clearly dominates the quenching process. As already postulated for generic phenolic derivatives, it proceeds through a polar intermediate complex which posses some component of charge-transfer character.Esterification of the carboxilic acid of tyrosine selectively decreases the contribution of the reactive step to the overall process of O₂(¹_g) quenching. An amide group in the same position does not produce noticiable changes in this sense. The presence of a highly deactivating nitro group in nuclear positions greatly diminishes the magnitude of both overall and reactive interactions.For all three, o-, m- and p-tyrosine the values of photooxidation quantum yields show an excellent parallelism with the rates of consumption of the — NH₂ group of the amino acid chain, upon sensitized irradiation. It could react, in the cases of 0- and m-tyrosine in a secondary, non photochemical, step.Abbreviations O ₂(³ _g ^– ) ground state triplet oxygen - O ₂(¹_g) singlet molecular oxygen - Tyr L tyrosine - TyrD tyrosine derivatives - Eos eosine - RB rose bengal - FFA furfuryl alcohol - DMA 9, 10-dimethyl anthracene     

The long-term ecology of the lost forests of La Laguna, Tenerife (Canary Islands)

Aim We report the first analysis of the long‐term ecology of Tenerife, in order to establish a pre‐colonization base‐line and to assess the effect of human activity and the role of climatic variation on vegetation during the Late Holocene. Location A former lake bed in the city of La Laguna (Tenerife, Canary Islands, Spain). Methods A sedimentary sequence of over 2 m was obtained from the former lake bed. Fossil pollen and microfossil charcoal concentrations were analysed. Radiocarbon dating of the sequence indicates that it spans approximately the last 4700 years. The pollen diagram was zoned using optimal splitting within psimpoll 4.25. Results Three pollen zones were differentiated: (1) in Zone L1 (c. 4700–2900 cal. yr bp ) a mixed forest was dominated by Quercus, Carpinus, Myrica and Pinus; (2) in Zone L2 (c. 2900–2000 cal. yr bp ) the laurel forest taxa increased, while Pinus, Juniperus and Phoenix declined; and (3) Zone L3 (c. 2000–400 cal. yr bp ) was characterized by the decline of Carpinus and Quercus and the abundance of laurel forest taxa (e.g. Myrica). Neither Carpinus nor Quercus was hitherto considered to be native to the Canary Islands. Their decline started c. 2000 years ago, coinciding with microfossil charcoal evidence of increased burning and with archaeological evidence for the first human settlement on Tenerife. Main conclusions Between c. 4700 and 2000 cal. yr bp , the composition of the forest in the valley of La Laguna was very different from what it is at present. In particular, Quercus and Carpinus appear to have been significant components, alongside components of the present‐day laurel forest, and the native pine (Pinus canariensis) forest and thermophilous woodland were also more prevalent in the region (but probably not within the lake basin itself) until 3000 cal. yr bp . The subsequent decline of Quercus and Carpinus led to the establishment of the present laurel forest in the region and a shift to more open vegetation types. These changes indicate that the aboriginal inhabitants of the islands, the Guanches, had a far more profound impact on the vegetation of Tenerife than hitherto realized.     

Increased phosphoinositide 3-kinase activity induces a lymphoproliferative disorder and contributes to tumor generation in vivo.

Alterations in the cell division:cell death ratio induce multiple autoimmune and transformation processes. Phosphoinositide 3-kinase (PI3K) controls cell division and cell death in vitro, but its effect on the function of the cellular immune system and on tumor formation in mammals is poorly characterized. Here we show that transgenic mice expressing in T lymphocytes an active form of PI3K derived from a thymic lymphoma, p65(PI3K), developed an infiltrating lymphoproliferative disorder and autoimmune renal disease with an increased number of T lymphocytes exhibiting a memory phenotype and reduced apoptosis. This pathology was strikingly similar to that described in mice exhibiting heterozygous loss of the tumor suppressor PTEN, a lipid and protein phosphatase. We show that overexpression of PTEN selectively blocks p65(PI3K)-induced 3T3 fibroblast transformation. Moreover, the early development of T cell lymphomas in p65(PI3K) Tg p53(-/-) mice indicated that PI3K contributes to tumor development. These observations demonstrate that constitutive activation of PI3K extends T cell survival in vivo, affects T cell homeostasis, and contributes to tumor generation, supporting a model in which selective increases in one type of PTEN substrate, the PI3K-derived lipid products, induce tumorigenesis. PI3K thus emerges as a potential target in autoimmune disease and cancer therapy.