Bacteria and Archaea community structure in the rumen microbiome of goats (Capra hircus) from the semiarid region of Brazil

Most studies present in the literature about the rumen microbiome have focused on cattle and sheep. This is the first report of the characterization of the bacterial and archaeal communities present in the liquid and solid-associated fractions of the rumen from free ranging Moxotó breed goats using 16S rRNA gene libraries. PCR was used to amplify the 16S rRNA gene with bacterial and archaeal universal primers and sequences from each library constructed were obtained. Sequences of Bacteria from the phyla Bacteroidetes and Firmicutes were predominant. The overall dominant classes in the rumen were Clostridia and Bacteroidia, which are known to play a role in plant fiber degradation in other ruminants. Unclassified Bacteria accounted for 4.7% of the liquid fraction sequences and 16.4% of the solid fraction sequences. From the archaeal libraries only sequences from the phylum Euryarcheota were identified and were assigned to the class Methanobacteria of the genera Methanobrevibacter and Methanosphaera. A group of Archaea not previously known to be associated with the rumen was identified: uncultured methanogens belonging to the "uncultured marine bacteria" groups II and III. The local water contained high salt concentrations and this may explain the presence of these groups in the Moxotó goat rumen.     

RF nonlinear interactions in living cells--II: detection methods for spectral signatures

The presence of harmonic products due to possible nonlinear interaction of amplitude modulated RF signals in living cells is best detected by using a cavity with high quality factor. Harmonic products generated by elementary oscillators can be trapped and accumulated in a cavity, permitting detection sensitivity much greater than in an open environment, where they would be radiated in all directions. The experimental method described herein is a systematic approach to detection of the non-Planck RF energy (if any) emitted by an exposed sample of living cells. Balzano and Sheppard [Balzano and Sheppard (2003): Bioelectromagnetics 24:473-482] classified the non-Planck RF emissions from living cells as coming from (1). nonlinear interactions and (2). inelastic interactions. Nonlinear harmonic products would appear in the band at twice the frequency of an amplitude modulated RF carrier. Inelastic interaction products resulting from the interaction between the incident RF energy and normally occurring mechanical vibrations are found in the band immediately adjacent to the carrier. Detection of the latter signals is difficult because of this close spectral proximity, for example, 1 part in 10(7) for 100 Hz modulation of a GHz carrier. Modern audio spectrum analyzers have excellent selectivity, providing 60 dB rejections only 2 kHz away from the carrier. By judicious selection of the amplitude modulation (AM) frequency, frequency of the RF carrier, and size of the biological sample, it is possible to achieve very high sensitivity (about -90 dBm) with commercially available instrumentation. The presence (or absence) of harmonics in the band adjacent to the amplitude modulated RF carrier would establish (or negate) the existence of coherent interactions between mechanical vibrations in the cell ensemble and the incident RF signal.     

Evaluation of four mathematical functions to describe scrotal circumference maturation in Nellore bulls

Four functions to characterize scrotal circumference (SC) growth in Nellore bulls were compared to identify which was the most suitable for biological interpretation. Nellore bulls (n = 532), born between September and December of 1992 to 1994 were used in the study. Measurements were made on fixed dates in January, April, July and October of each year. At the time of SC measurements, the ages of the bulls ranged from 200 to 1300 d. The functions used to describe the data were: Brody, SC = A (1 - B exp -kt); Logistic, SC = A/(1 + B exp -kt); Gompertz, SC = A exp(-B exp -kt) and Richards SC = A (1 + B exp -kt)m, where SC is the scrotal circumference at t days of age, A is the estimated SC at maturity, B is the integration constant established by the initial values of SC and t, k is the maturity constant, which equals the ratio between the maximum rate of growth and SC at maturity; m is the inflection point parameter for Richards function, which did not converge. The Brody, Gompertz and Logistic functions fitted the data in a similar fashion, with similar values for the statistics EMS and R2, and they reached convergence with similar computational costs. The Logistic function presented the best pattern of average prediction error, and therefore, it was selected for biological interpretation. For the Logistic function, estimated SC at maturity (A) was 37.95 cm at 72 mo of age. The maturity index (k) was .11 cm, and the inflection point (time of maximum growth) was reached at 13.09 mo of age at an average SC of 18.97 cm.     

Synthesis, structural characterization, and photo and electroluminescence of a novel terbium(III) complex: {Tris(acetylacetonate) [1,2,5]thiadiazolo[3,4-f][1,10]phenanthroline}terbium(III)

Lanthanides ion complexes have been intensively investigated as light emitting materials due to their interesting photophysical properties, such as narrow line luminescence with a long lifetime, large Stokes shift and high luminescence quantum efficiency. Here we report the synthesis, structural and photophysical properties of a new Tb(III) complex. This complex showed strong photoemission of green light both in solution and in the solid state as well as the characteristic emission lines of the Tb(III) ion. The electroluminescence properties of the complex were also studied and we obtained bright green light emission through the use of a co-deposited structure. The fabricated device showed a typical diode behavior with a low threshold bias voltage (around 10 V).     

Sequencing,cloning, and heterologous expression of cyclomaltodextrin glucanotransferase of Bacillus firmus strain 37 in Bacillus subtilis WB800

Bacillusfirmus strain 37 produces the cyclomaltodextrin glucanotransferase (CGTase) enzyme and CGTase produces cyclodextrins (CDs) through a starch cyclization reaction. The strategy for the cloning and expression of recombinant CGTase is a potentially viable alternative for the economically viable production of CGTase for use in industrial processes. The present study used Bacillus subtilis WB800 as a bacterial expression host for the production of recombinant CGTase cloned from the CGTase gene of B. firmus strain 37. The CGTase gene was cloned in TOPO-TA^® plasmid, which was transformed in Escherichia coli DH5α. The subcloning was carried out with pWB980 plasmid and transformation in B. subtilis WB800. The 2xYT medium was the most suitable for the production of recombinant CGTase. The enzymatic activity of the crude extract of the recombinant CGTase of B. subtilis WB800 was 1.33 µmol β-CD/min/mL, or 7.4 times greater than the enzymatic activity of the crude extract of CGTase obtained from the wild strain. Following purification, the recombinant CGTase exhibited an enzymatic activity of 157.78 µmol β-CD/min/mL, while the activity of the CGTase from the wild strain was 9.54 µmol β-CD/min/mL. When optimal CDs production conditions for the CGTase from B. firmus strain 37 were used, it was observed that the catalytic properties of the CGTase enzymes were equivalent. The strategy for the cloning and expression of CGTase in B. subtilis WB800 was efficient, with the production of greater quantities of CGTase than with the wild strain, offering essential data for the large-scale production of the recombinant enzyme.

     

Synchronous fruiting and common seed dispersers of two endemic columnar cacti in the Caatinga,a dry forest in Brazil

This study evaluates the fruiting phenology, fruit traits, and seed dispersal in two Pilosocereus (Cactaceae) species that are widely distributed in Caatinga vegetation. We monitored the fruiting phenology of Pilosocereus gounellei and Pilosocereus chrysostele on a monthly basis for a period of 4 years (45 months from March 2009 to November 2012), including 30 individuals of each species. We also carried out focal observations, captured dispersers, and conducted germination tests, to identify the effective seed dispersers of these species. Both species exhibited sub-annual fruiting patterns and high fruiting synchrony index (O _jk 0.62), with peaks occurring from February to May for P. gounellei and February to April for P. chrysostele. In all, 248 visits by seven bird and two lizard species were recorded for P. gounellei, and 104 visits by five bird species were recorded for P. chrysostele. The two species shared five seed dispersers. The finch Lanio pileatus was the most frequently visiting bird species. The number of visits to the fruits of P. gounellei was higher than to the fruits of P. chrysostele. Passage of seeds through the digestive tracts of all bird species significantly increased the germination rate for P. chrysostele, whereas for P. gounellei, the birds Forpus xanthopterygius and L. pileatus decreased germination rates. Sub-annual fruiting patterns, similar fruit morphology, and high synchronous fruiting are factors that favor resource sharing among the dispersers of Pilosocereus in the Caatinga.     

Knockin’ on pHeaven’s Door: A Fast and Reliable High-Throughput Compatible Zero-Background Cloning Procedure

We created a pair of vectors allowing simple and efficient molecular cloning of any gene of interest with minimal consumption of time, labor and material. This system is applicable for standard molecular cloning, high-throughput cloning and generation of fusion protein libraries as well as for more complex gene assembly purposes. Also, this zero-background procedure allows going from cDNA to gene expression analysis in a defined vector in <2 days.     

Immunopositivity for Siglec-15 in gastric cancer and its association with clinical and pathological parameters

The sialic acid-binding immunoglobulin-type lectin Siglec-15 is a promising target for cancer immunotherapy in several tumor types. The present study aimed to investigate Siglec-15 expression in gastric cancer (GC) patient tissues and to evaluate its clinical value. Siglec-15 expression was evaluated by immunohistochemistry in 71 patients. Siglec-15 staining was observed in tumor cells of 53 (74.64%) patients, with significant association with histologic classification and angiolymphatic invasion (p<0.05). Immunohistochemistry analysis also detected Siglec-15 in tumor-associated stroma cells (macrophages/myeloid cells). There was no significant association with outcome parameters. Siglec-15 expression in well differentiated histological GC tissues and in the tumor microenvironment are potential targets to be further investigated as a novel prognostic factor for GC.Key words: Siglec, sialic acid, immunohistochemistry, clinical-pathological analyses