Selective Permeation and Organic Extraction of Recombinant Green Fluorescent Protein (gfp uv ) from Escherichia coli

Background  

Transformed cells of Escherichia coli DH5-α with pGFPuv, induced by IPTG (isopropyl-β-d-thiogalactopyranoside), express the green fluorescent protein (gfp _uv ) during growth phases. E. coli subjected to the combination of selective permeation by freezing/thawing/sonication cycles followed by the three-phase partitioning extraction (TPP) method were compared to the direct application of TPP to the same culture of E. coli on releasing gfp _uv from the over-expressing cells.     

Influence of training in the dark or light phase on physiologic and metabolic parameters of Wistar rats submitted to aerobic exercise

Light and dark phase training may influences rodents’ physiologic parameters because these animals have nocturnal habits. Thus, we verify the effects of the training in different photoperiods on metabolism and corporal composition of rats. Eighteen rats were divided into three groups – G1: non-trained; G2: trained in the light phase; G3: trained in the dark phase. Rats were allowed to swim for 60 min, five times per week during six weeks. Trained animals presented a smaller weight gain and fat percentage in carcass. Rats of G3 increased gastrocnemius relative weight. The adipocyte diameter of G3 rats was smaller than the other groups. The levels of the total cholesterol, low-density proteins, and triacylglycerols were decreased in animals of G2 while the glycemia was increased. Training in light phase provided more alterations in the blood biochemical profile while the training in the dark increased the gastrocnemius weight and decreased the diameter of the adipocyte.     

Breast and Bottle Feeding as Risk Factors for Dental Caries: A Systematic Review and Meta-Analysis

Understanding the role that breastfeeding and bottle feeding play in the development of dental caries during childhood is essential in helping dentists and parents and care providers prevent the disease, and also for the development of effective public health policies. However, the issue is not yet fully understood. The aim of this systematic review and meta-analysis was to search for scientific evidence in response to the question: Do bottle fed children have more dental caries in primary dentition than breastfed children? Seven electronic databases and grey literature were used in the search. The protocol number of the study is PROSPERO CRD 42014006534. Two independent reviewers selected the studies, extracted data and evaluated risk of bias by quality assessment. A random effect model was used for meta-analysis, and the summary effect measure were calculated by odds ratio (OR) and 95% CI. Seven studies were included: five cross-sectional, one case-control and one cohort study. A meta-analysis of cross-sectional studies showed that breastfed children were less affected by dental caries than bottle fed children (OR: 0.43; 95%CI: 0.23–0.80). Four studies showed that bottle fed children had more dental caries (p<0.05), while three studies found no such association (p>0.05). The scientific evidence therefore indicated that breastfeeding can protect against dental caries in early childhood. The benefits of breastfeeding until age two is recommended by WHO/UNICEF guidelines. Further prospective observational cohort studies are needed to strengthen the evidence.     

Successful Treatment of Canine Sporotrichosis with Terbinafine: Case Reports and Literature Review

Sporotrichosis occurs worldwide, and the metropolitan region of Rio de Janeiro, Brazil, is a main endemic area, with a large number of human and animal cases in the last 19 years. This mycosis is more frequently described in cats rather than in dogs. There are a limited number of oral antifungal agents for the treatment of sporotrichosis in animals. In this context, the effectiveness of terbinafine in the treatment of sporotrichosis in humans, as well as the promising results of in vitro susceptibility tests, inspired us to use this drug in the therapy of this mycosis in dogs. We reported for the first time the use of terbinafine in the treatment of two dogs with sporotrichosis caused by Sporothrix brasiliensis. Moreover, we provided an overview of therapeutic features of canine sporotrichosis cases reported since the 1960s. One of the dogs presented the fixed cutaneous form of the disease, while the other patient presented hyperemia of the nasal mucosa and respiratory signs only. Terbinafine showed high antifungal activity in vitro against the canine Sporothrix isolates. The dogs were successfully treated with terbinafine, with remission of all clinical signs initially presented. The current reports indicate that this drug can emerge as a therapeutic option for canine sporotrichosis.     

Total phenolics,resveratrol content and antioxidant activity of seeds and calluses of pinto peanut (<Emphasis Type="Italic">Arachis pintoi</Emphasis> Krapov. &amp; W.C. Greg.)

Arachis pintoi is a peanut species native to Brazil, which is cultivated in many countries for animal forage, soil cover, landscaping, and recovery of degraded areas. Tissue culture studies for this species have been focused in plant production, whereas works on in vitro secondary metabolites production are scarce. The goal of the present work was to establish callus cultures from different seed explants of A. pintoi, aiming at evaluating the potential for metabolites production and antioxidant activity. Embryonic axes, leaflets, and cotyledons were cultured on solidified MS medium supplemented with picloram (PIC), 2,4-dichlorophenoxyacetic acid (2,4-D), thidiazuron (TDZ) or different combinations of 6-benzyladenine (BA) and α-naphthaleneacetic acid (NAA), under light or dark conditions. Friable calluses with a high biomass (4.3?±?0.3 g FW per callus) were obtained from embryonic leaflets cultured on medium supplemented with 17.6 µM BA plus 5.4 µM NAA, in the dark. Cotyledons and embryonic axes cultured in the presence of 4.4 µM BA combined with 10.8 µM NAA formed heterogeneous calluses with a compact base and a large friable surface. Trans-resveratrol and other stilbenes that were not found in seeds were detected in callus extracts, especially those originated from cotyledons, although these materials showed lower total phenolic contents (TPC) when compared with seeds with and without testa, as well as cotyledons. Extracts from seeds with testa and from calluses derived from cotyledons and embryonic axes showed the highest EC₅₀ in DPPH assays. No correlation between TPC, trans-resveratrol and antioxidant activity was observed.     

Temporal selectivity for complex signals by single neurons in the torus semicircularis of Pleurodema thaul (Amphibia: Leptodactylidae)

Responses of auditory neurons in the torus semicircularis (TS) of Pleurodema thaul, a leptodactylid from Chile, to synthetic stimuli having diverse temporal patterns and to digitized advertisement calls of P. thaul and three sympatric species, were recorded to investigate their temporal response selectivities. The advertisement call of this species consists of a long sequence of sound pulses (a pulse-amplitude-modulated, or PAM, signal) having a dominant frequency of about 2000 Hz. Each of the sound pulses contains intra-pulse sinusoidal-amplitude-modulations (SAMs). Synthetic stimuli consisted of six series in which the following acoustic parameters were systematically modified, one at a time: PAM rate, pulse duration, number of pulses, and intra-pulse SAM rate. The carrier frequency of these stimuli was set at the characteristic frequency (CF) of the isolated units (n = 47). Response patterns of TS units to synthetic call variants reveal different degrees of selectivities for each of the temporal variables, with populations of neurons responding maximally to specific values found in the advertisement call of this species. These selectivities are mainly shaped by neuronal responsiveness to the overall sound energy of the stimulus and by the inability of neurons to discharge to short inter-pulse gaps. Accepted: 30 October 1996     

Characterization of Zea mays endosperm C-24 sterol methyltransferase: one of two types of sterol methyltransferase in higher plants

We report the characterization of a higher-plant C-24 sterol methyltransferase by yeast complementation. A Zea mays endosperm expressed sequence tag (EST) was identified which, upon complete sequencing, showed 46% identity to the yeast C-24 methyltransferase gene (ERG6) and 75% and 37% amino acid identity to recently isolated higher-plant sterol methyltransferases from soybean and Arabidopsis, respectively. When placed under GAL4 regulation, the Z. mays cDNA functionally complemented the erg6 mutation, restoring ergosterol production and conferring resistance to cycloheximide. Complementation was both plasmid-dependent and galactose-inducible. The Z. mays cDNA clone contains an open reading frame encoding a 40 kDa protein containing motifs common to a large number of S-adenosyl-L-methionine methyltransferases (SMTs). Sequence comparisons and functional studies of the maize, soybean and Arabidopsis cDNAs indicates two types of C-24 SMTs exist in higher plants.