PCR screening reveals abundance of bovicin-like bacteriocins among ruminal Streptococcus spp. isolated from beef and dairy cattle.

AIMS: To investigate the inhibitory activity and the distribution of biosynthetic genes encoding bovicin-like bacteriocins among ruminal Streptococcus isolated from beef and dairy cattle. METHODS AND RESULTS: Most isolates were classified as Streptococcus equinus and Streptococcus lutetiensis based on 16S rRNA sequencing. The antimicrobial activity of 150 ruminal streptococci isolated from beef and dairy cattle were tested by deferred inhibition assays and their genetic diversity was characterized by BOX-PCR. The frequency of biosynthetic genes associated with the biosynthesis of bovicin-like bacteriocins (bovicin HC5 and bovicin 255) was investigated by PCR screening. Approximately 33% of the ruminal streptococci isolated from Nellore heifers showed inhibitory activity in vitro with the majority harbouring genes for bacteriocin biosynthesis. In contrast, streptococci from Holstein cows showed limited inhibitory activity and a lower frequency of bacteriocin biosynthetic genes. CONCLUSIONS: Streptococcus from the rumen of beef and dairy cattle exhibit remarkable differences in inhibitory activity and distribution of genes associated with the biosynthesis of prototypical bovicins (bovicin HC5 and bovicin 255). SIGNIFICANCE AND IMPACT OF THE STUDY: Our findings demonstrate that bovicin HC5 is distributed among ruminal streptococci from different breeds of cattle. The high degree of conservation of the bovicin HC5 structural gene among strains of ruminal streptococci suggests that random genetic drift is not a dominant force in the evolution of this bacteriocin.

Eugenol influences the expression of messenger RNAs for superoxide dismutase and glutathione peroxidase 1 in bovine secondary follicles cultured in vitro.

This study aimed to investigate the effects of eugenol on growth, viability, antrum formation and mRNA expression of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase 1 (GPX1) and peroxiredoxin 6 (PRDX6) in bovine secondary follicles cultured in vitro. To this end, bovine ovaries were collected from a local slaughterhouse and in the laboratory the follicles were isolated from the ovarian cortex. The follicles were then cultured in TCM-199+ alone or supplemented with different concentrations of eugenol (0.5, 5.0 and 50.0 µM). Follicular diameters and antrum formation were evaluated on days 0, 6, 12 and 18. Viability analysis was performed using calcein and ethidium homodimer. Real-time PCR was used to quantify mRNA levels for SOD, CAT, GPX1 and PRDX6 in cultured follicles. Follicular diameters and mRNA levels in follicles cultured in vitro were compared using analysis of variance and Kruskal-Wallis tests, while follicular survival and antrum formation were compared using the chi-squared test (P < 0.05). The results showed that secondary follicles cultured with eugenol maintained similar morphology and viability to follicles cultured in the control group. A progressive increase in follicular diameter was observed between days 0 and 12 for all treatments, except for follicles cultured with 50 µM eugenol. Eugenol (5.0 and 50.0 µM) increased mRNA levels for GPX1 in cultured follicles, but 0.5 µM eugenol reduced mRNA levels for SOD. The addition of eugenol did not influence mRNA expression for CAT and PRDX6. In conclusion, eugenol supplementation reduces mRNA levels for SOD and increases mRNA levels of GPX1 in bovine secondary follicles cultured in vitro.

Soliton-like structures in the spectrum and the corresponding eigenstates morphology for the quantum desymmetrized Sinai billiard.

By continuously varying certain geometric parameters γ of the totally desymmetrized quantum Sinai billiard, we study the formation of the so-called soliton-like structures in the spectra of the resulting family of systems. We present a detailed characterization of the eigenstate ψn morphologies along such structures. Usually, scarring and bouncing ball mode states are expected to fully explain the solitons. However, we show that they do not exhaust all the possibilities. States with strong resemblance to very particular solutions of the associated integrable case ( 45°- 45° right triangle) also account for the ψn's. We argue that for the emergence of the solitons, in fact, there must be an interplay between the spatial localization properties of the soliton-related ψn's and the rescaling properties of the billiards with γ. This is illustrated, e.g., by comparing the behavior of the eigenwavelengths along the solitons and the billiard size dependence on γ. Considerations on how these findings could extend to other type of billiards are also briefly addressed.

On the Use of Embedded Fiber Optic Sensors for Measuring Early-Age Strains in Concrete.

Detailed information about concrete behavior in real structures is an important issue in controlling its performance during its service life, and the use of embedded sensors to obtain desired information such as temperature, onset of the cracking process and evolution of strains, has gained the attention of the building concrete industry. Data obtained using this technology can provide valuable information for decision making about the need for corrective interventions that can ensure the integrity and safety of concrete structures for long period of time. This paper presents a review of the current state-of-the-art of embedded fiber optic sensors used to assess concrete information of a wide range of aspects, comprising: existing alternative technologies, characteristics and advantages, practical applications and future developments. Complementarily, the work presents preliminary results of the use of fiber optic sensors to automatically and continuously perform expansion readings of AAR in concrete elements that facilitate both the storage-with elimination of the usual interruptions for manual readings-and the availability of continuous results of expansion data that are not possible to obtain with usual AAR tests code reading recommendations.

Obesity-related knowledge and body mass index: a national survey in Portugal.

PURPOSE: Obesity-related knowledge predicts weight control but previous studies only comprise individuals with excessive weight and assess very specific aspects of knowledge. This study aims to evaluate several domains of obesity-related knowledge according to the body mass index (BMI) in a representative sample of Portuguese-speaking dwellers in mainland Portugal. METHODS: A sample of 1624 participants aged 16-79 years was analyzed. Eighteen questions comprising eight obesity domains were asked: prevalence, physical activity, number of calories, adiposity location, causes, diagnosis, treatment and consequences; each one was dichotomized into correct/incorrect knowledge. RESULTS: The majority of Portuguese-speaking dwellers recognized the benefits of physical activity, the risks of abdominal obesity and most consequences of excessive weight, independently of their BMI. However, knowledge gaps were identified regarding prevalence, calories and BMI diagnosis. BMI influenced specific obesity-related knowledge: participants with a normal BMI knew the BMI formula more often, identified 22 as normal BMI more frequently and had the highest proportion of correct knowledge regarding the number of calories an adult should eat; obese individuals identified natural products as not being good treatments for obesity more often. After adjusting for age, sex and educational level, obese individuals identified natural products and supplements as not being good obesity treatments more often. CONCLUSIONS: Obesity-related knowledge gaps (prevalence, calories, and diagnosis) were identified among Portuguese adults. Moreover, correct knowledge does not necessarily translate into a healthier BMI. Besides the dissemination of accurate information, public health interventions should focus on the transfer of knowledge to behaviors that will guarantee better weight management. EVIDENCE-BASED MEDICINE RANKINGS: Level V: Opinions of respected authorities, based on descriptive studies, narrative reviews, clinical experience, or reports of expert committees.

Oral potentially malignant disorders: clinical-pathological study of 684 cases diagnosed in a Brazilian population.

BACKGROUND: The frequency and distribution of oral potentially malignant disorders (OPMD) may vary among different populations. The aim of the present study was to evaluate the clinical-pathological characteristics of OPMD diagnosed in a Brazilian oral pathology laboratory over a period of 11 years. MATERIAL AND METHODS: All cases diagnosed as leukoplakia, speckled leukoplakia, erythroplakia, and actinic cheilitis from 2005 to 2015 were analyzed. Clinical information was obtained from laboratory forms and histological information was obtained from histological slides stained with hematoxylin and eosin. RESULTS: the final sample was comprised of 684 cases, of which 292 were males and 392 were females. The mean age was 58 years. The anatomical site most often involved was the lateral border of the tongue (23%), followed by the lower lip (20%), and by the buccal mucosa/vestibule (18%). Leukoplakia accounted for 82% of the sample (564 cases). The mean size of the leukoplakia and speckled leukoplakia lesions was 13 mm (ranging from 1 to 100 mm) and 15 mm (ranging from 5 to 30 mm), respectively (p=0.460). Males reported smoking and drinking alcohol more frequently than females (p=0.001 and p<0.0001, respectively). In half of the cases, dysplasia was not histologically detected, while slight dysplasia was detected in 28% of the cases. The lesions from patients aged from 41 to 80 years presented moderate and severe dysplasia more often than lesions from patients in other age groups. CONCLUSIONS: OPMD were more common in females in their sixties. Females were more frequently affected in all anatomical sites, except for the lips. Leukoplakia lesions were the most common OPMD, followed by actinic cheilitis. The lateral border of the tongue was the most affected anatomical site. OPMD located in the floor of the mouth/sublingual mucosa and lesions from older patients presented severe epithelial dysplasia with greater frequency.

Metformin and Breast Cancer: Molecular Targets.

Metformin has been the first-line drug for the treatment of type II diabetes mellitus for decades, being presently the most widely prescribed antihyperglycemic drug. Retrospective studies associate the use of metformin with a reduction in cancer incidence and cancer-related death. However, despite extensive research about the molecular effects of metformin in cancer cells, its mode of action remains controversial. The major molecular targets of metformin include complex I of the mitochondrial electron transport chain, adenosine monophosphate (AMP)-activated protein kinase (AMPK), and mechanistic target of rapamycin complex 1 (mTORC1), but AMPK-independent effects of metformin have also been described. Breast cancer is one of the leading causes of cancer-related morbidity and mortality among women worldwide. Several studies have reinforced a link between breast cancer risk and diabetes. Moreover, metformin significantly reduces breast cancer risk, compared to patients who are not using metformin and is independent of diabetes status. In this review, we summarize the current molecular evidence to elucidate metformin's mode of action against breast cancer cells.

The application of microbeads to microfluidic systems for enhanced detection and purification of biomolecules.

This paper describes microbead-based microfluidic systems. Several aspects of bead assays in microfluidics make them advantageous for bioassays in simple microchannels, including enhanced surface-to-volume ratio, improved molecular recognition reaction efficiency, and the wide range of surface functionalization available with commercial microbeads. Two-level SU-8 molds are used to fabricate PDMS microchannels that can hydrodynamically trap different types of microbeads, with characteristic dimensions of tens of microns. The use of these microbead-based microfluidic systems in the biosensing of antibodies, toxins and nucleic acids, as well as in antibody purification will be presented and discussed in this paper.

Bisoprolol-induced thrombocytopenia: A case report.

WHAT IS KNOWN AND OBJECTIVE: Thrombocytopenia, not associated with bone marrow primary disease, is a common clinical problem. The possibility of drug-induced thrombocytopenia must be considered, especially in hospitalized patients. Drugs can cause thrombocytopenia by several mechanisms including direct bone marrow or other organ toxicity, and immune reactions. CASE DESCRIPTION: We describe a patient presenting with thrombocytopenia likely related to bisoprolol. WHAT IS NEW AND CONCLUSION: We report a case of bisoprolol-induced thrombocytopenia which resolved with drug discontinuation and steroid therapy. We review the mechanisms involved in drug-induced immune thrombocytopenia.

A biomolecular recognition approach for the functionalization of cellulose with gold nanoparticles.

Materials with new and improved functionalities can be obtained by modifying cellulose with gold nanoparticles (AuNPs) via the in situ reduction of a gold precursor or the deposition or covalent immobilization of pre-synthesized AuNPs. Here, we present an alternative biomolecular recognition approach to functionalize cellulose with biotin-AuNPs that relies on a complex of 2 recognition elements: a ZZ-CBM3 fusion that combines a carbohydrate-binding module (CBM) with the ZZ fragment of the staphylococcal protein A and an anti-biotin antibody. Paper and cellulose microparticles with AuNPs immobilized via the ZZ-CBM3:anti-biotin IgG supramolecular complex displayed an intense red color, whereas essentially no color was detected when AuNPs were deposited over the unmodified materials. Scanning electron microscopy analysis revealed a homogeneous distribution of AuNPs when immobilized via ZZ-CBM3:anti-biotin IgG complexes and aggregation of AuNPs when deposited over paper, suggesting that color differences are due to interparticle plasmon coupling effects. The approach could be used to functionalize paper substrates and cellulose nanocrystals with AuNPs. More important, however, is the fact that the occurrence of a biomolecular recognition event between the CBM-immobilized antibody and its specific, AuNP-conjugated antigen is signaled by red color. This opens up the way for the development of simple and straightforward paper/cellulose-based tests where detection of a target analyte can be made by direct use of color signaling.

Skeletal anomalies in reared Senegalese sole Solea senegalensis juveniles: a radiographic approach.

Reared Senegalese sole Solea senegalensis Kaup show a high incidence of vertebral anomalies; however, little is known about its skeletal anomaly profile in the later farming phases. The purpose of this study was to provide a detailed description and quantification of the most common skeletal anomalies in reared Senegalese sole in the juvenile stage by means of computed radiography. A total of 374 Senegalese sole were classified according to the external morphology of the fish as normal or altered and then radiographed in latero-lateral and in dorso-ventral projections. Radiographic evaluation of anomalies focused especially on vertebral body anomalies (VBA) and vertebral column deviations (VCD). The 2 orthogonal projections provided a more complete visualization of the skeleton. Approximately 75% of the individuals showed at least 1 anomaly, while VBA and/or VCD were detected in 48.9% of the specimens. Regarding external morphology, 88% of the fish were categorized as normal, although about 72% of these normal fish displayed abnormalities in radiographies. The most frequent anomalies consisted of deformations of the caudal complex plates (hypurals, parhypural and epural), preurals and caudal vertebrae. Scoliosis was the most prevalent among VCD, affecting the caudal area in almost 15% of the individuals. The anomaly profile at the juvenile stages showed some differences compared to what has been reported previously in earlier stages of development. In light of these results, further investigation into the progression of skeletal anomalies over time and the causative factors at later stages is required.

New insight on vertebral anomalies in cultured Senegalese sole (Solea senegalensis, Kaup) at early stages of development.

Senegalese sole (Solea senegalensis, Kaup) is a promising flatfish species in aquaculture. However, skeletal anomalies are still a great concern in sole farming. Investigation of this issue is crucial to improving larval quality and optimizing production. The aim of this study was to thoroughly assess anomalies in the rachis of reared sole at early developmental stages. Sole (n = 507) were sampled at 31 or 32 days after hatching (dah). The specimens were stained with alcian blue and alizarin red and evaluated for the detection of vertebral deformities. Most fish presented 9:34:3 vertebrae in abdominal, caudal and caudal complex regions, respectively. Remarkably, all specimens showed at least one spinal anomaly. Alterations of neural/haemal elements, as well as deformities of hypurals, parhypural and epural, were recurrent. Vertebral body anomalies and/or vertebral column deviations were identified in 52% of the individuals. Vertebral deformations and fusions were common, especially in caudal complex. 'Minor' anomalies were predominant, and some of the detected disorders might be a result of non-/low-pathological processes. These results contribute a new insight into the main skeletal anomalies affecting cultured sole larvae. Further research is required to determine their impact on fish welfare and external appearances at commercial stages.

Selecting black-spot resistant papaya genotypes derived from backcrossing and hybrids.

Papaya crop is important to Brazilian agribusiness. However, the expansion of papaya cultivation in the country is affected by the absence of commercial cultivars presenting good disease resistance. The black-spot caused by the fungus Asperisporium caricae is the most damaging foliar disease affecting Brazilian papaya crops. The use of genetically resistant cultivars is a promising strategy to reduce the dependence of papaya crops on fungicides. A field split-plot experiment was carried out in the municipality of Linhares, Espírito Santo State, and included 20 hybrids derived from the cross between 14 superior lines and four elite genotypes ('SS72/12', 'SEKATI', 'JS/12' and '41/7'), two commercial cultivars ('Golden' and 'Tainung 01'), and the superior line '36/7', which were evaluated for resistance to black-spot in the fruits and leaves. The treatments were arranged in a randomized block design with six repetitions of three plants per plot. The incidence and severity of black spot in the fruits and leaves were evaluated at three different times in the 2015-2016 crop season. Lines 4, 9, 21, and the parent SEKATI were notable for their capacity to reduce disease severity in the leaves and fruits. Lines 1, 2, 9, 16, and 19, and the parents 'SEKATI' and 'SS-72/12' had reduced disease incidence in their fruits. The most resistant hybrids 'SS-72/12 X 4', 'SS-72/12 X 6', 'SEKATI X 1', 'SEKATI X 2', 'SEKATI X 6', 'SEKATI X 9', and 'SEKATI X 20' presented negative heterosis values for improved black-spot resistance. The current study allowed the selection of black-spot resistant genotypes and hybrids, which presented a significantly reduced disease index in the field.

BCR-ABL1 transcript types showed distinct laboratory characteristics in patients with chronic myeloid leukemia.

In chronic myeloid leukemia (CML) two main types of messenger RNA (e14a2 and e13a2) can be produced by BCR-ABL1 gene rearrangement. Due to conflicting results, the clinical value of these transcripts remains controversial. The aim of this study was to identify associations of e14a2 and e13a2 transcripts with laboratory variables and also the response to treatment. This study included 203 adult patients with CML treated with Imatinib as first-line drug in a reference hematology center in Northeast Brazil. Clinical and laboratory data were obtained after informed consent. Samples were collected for RNA extraction and analyzed by reverse transcription-polymerase chain reaction (PCR), according to the international protocol BIOMED-1. The LeukemiaNet 2013 criteria were used to establish the molecular response. The frequency distribution of the BCR-ABL1 transcripts was e14a2 (64%), e13a2 (34%), and double positives (2%). The results showed a statistically significant association of the e14a2 transcript type with thrombocytosis (P = 0.0005) and the e13a2 with higher leukocyte count (P = 0.0491). In a subgroup of 44 patients, the molecular response to treatment with Imatinib was assessed by quantitative PCR at 3 months (BCR-ABL1 ≤ 10%), 6 months (BCR-ABL1 ≤ 1%), or 12 months (BCR-ABL1 ≤ 0.1%). Although patients with the transcript e14a2 showed higher frequency of good responses than patients with the transcript e13a2, this difference was not statistically significant. In agreement with published data, our results showed association of the BCR-ABL1 transcript e14a2 with thrombocytosis and the BCR-ABL1 transcript e13a2 with higher leukocytosis in patients with chronic myeloid leukemia.

Raw water clarification by flotation with microbubbles and nanobubbles generated with a multiphase pump.

Raw water clarification by flotation was studied by injecting air into a centrifugal multiphase pump to generate microbubbles (MBs) and nanobubbles (NBs). Measurements of gas dispersion parameters were performed and optimal conditions were obtained using a pump pressure of 4 bar. Values showed a bubble Sauter diameter of 75 µm, an air holdup of 1.2%, a bubble surface area flux of 34 s-1 and an NB concentration of 1 × 108 NBs mL-1 (measuring 220 nm). Then, a study compared flotation with bubbles formed with the multiphase pump (F-MP) to lamellar settling at the clarification stage of a water treatment plant (WTP), in Brazil. The F-MP showed a higher separation efficiency at high hydraulic loads (9-15 m h-1), even without the use of a polymer, reaching 2 NTU (10-25 NTU raw water feed), which was much lower than the technical goal of the WTP (5 NTU). The results and the technical aspects are discussed, and it is concluded that the employment of MBs and NBs with pumps widens new research lines and applications in modern flotation.

Separation of emulsified crude oil in saline water by flotation with micro- and nanobubbles generated by a multiphase pump.

The flocculation-column flotation with hydraulic loading (HL, >10 m h-1) was studied for the treatment of oil-in-water emulsions containing 70-400 mg L-1 (turbidity = 70-226 NTU) of oil and salinity (30 and 100 g L-1). A polyacrylamide (Dismulgan, 20 mg L-1) flocculated the oil droplets, using two floc generator reactors, with rapid and slow mixing stages (head loss = 0.9 to 3.5 bar). Flotation was conducted in two cells (1.5 and 2.5 m) with microbubbles (MBs, 5-80 µm) and nanobubbles (NBs, 50-300 nm diameter, concentration of 108 NBs mL-1). Bubbles were formed using a centrifugal multiphase pump, with optimized parameters and a needle valve. The results showed higher efficiency with the taller column reducing the residual oil content to 4 mg L-1 and turbidity to 7 NTU. At high HL (27.5 m h-1), the residual oil concentrations were below the standard emission (29 mg L-1), reaching 18 mg L-1. The best results were obtained with high concentration of NBs (apart from the bigger bubbles). Mechanisms involved appear to be attachment and entrapment of the NBs onto and inside the flocs. Thus, the aggregates were readily captured, by bigger bubbles (mostly MBs) aiding shear withstanding. Advantages are the small footprint of the cells, low residence time and high processing rate.

Oral yeast colonization throughout pregnancy.

BACKGROUND: Recent studies suggest that placenta may harbour a unique microbiome that may have origin in maternal oral microbiome. Although the major physiological and hormonal adjustments observed in pregnant women lead to biochemical and microbiological modifications of the oral environment, very few studies evaluated the changes suffered by the oral microbiota throughout pregnancy. So, the aim of our study was to evaluate oral yeast colonization throughout pregnancy and to compare it with non-pregnant women. MATERIAL AND METHODS: The oral yeast colonization was assessed in saliva of 30 pregnant and non-pregnant women longitudinally over a 6-months period. Demographic information was collected, a non-invasive intra-oral examination was performed and saliva flow and pH were determined. RESULTS: Pregnant and non-pregnant groups were similar regarding age and level of education. Saliva flow rate did not differ, but saliva pH was lower in pregnant than in non-pregnant women. Oral yeast prevalence was higher in pregnant than in non-pregnant women, either in the first or in the third trimester, but did not attain statistical significance. In individuals colonized with yeast, the total yeast quantification (Log10CFU/mL) increase from the 1st to the 3rd trimester in pregnant women, but not in non-pregnant women. CONCLUSIONS: Pregnancy may favour oral yeast growth that may be associated with an acidic oral environment.

Clinical Efficacy and Safety of Ixekizumab for Treatment of Psoriasis.

Psoriasis is a common, chronic, inflammatory skin disorder with a physical and emotional burden. Emerging evidence suggests that IL17-A is a key cytokine in the immunopathogenesis of psoriasis. Ixekizumab is a humanized IgG4 monoclonal antibody that acts by neutralizing IL-17A. Data from Phase I-III studies reveal that ixekizumab is highly effective in treating patients with moderate-to-severe plaque psoriasis. A large proportion of patients receiving ixekizumab achieved or maintained complete or near complete resolution of psoriatic lesions with an acceptable safety profile through week 60. These remarkable results introduce a paradigm shift in the medically management of psoriasis, where complete or almost completely clear skin becomes the new therapeutic goal.

Coconut genome size determined by flow cytometry: Tall versus Dwarf types.

Coconuts (Cocos nucifera L.) are tropical palm trees that are classified into Tall and Dwarf types based on height, and both types are diploid (2n = 2x = 32 chromosomes). The reproduction mode is autogamous for Dwarf types and allogamous for Tall types. One hypothesis for the origin of the Dwarf coconut suggests that it is a Tall variant that resulted from either mutation or inbreeding, and differences in genome size between the two types would support this hypothesis. In this study, we estimated the genome sizes of 14 coconut accessions (eight Tall and six Dwarf types) using flow cytometry. Nuclei were extracted from leaf discs and stained with propidium iodide, and Pisum sativum (2C = 9.07 pg DNA) was used as an internal standard. Histograms with good resolution and low coefficients of variation (2.5 to 3.2%) were obtained. The 2C DNA content ranged from 5.72 to 5.48 pg for Tall accessions and from 5.58 to 5.52 pg for Dwarf accessions. The mean genome sizes for Tall and Dwarf specimens were 5.59 and 5.55 pg, respectively. Among all accessions, Rennel Island Tall had the highest mean DNA content (5.72 pg), whereas West African Tall had the lowest (5.48 pg). The mean coconut genome size (2C = 5.57 pg, corresponding to 2723.73 Mbp/haploid set) was classified as small. Only small differences in genome size existed among the coconut accessions, suggesting that the Dwarf type did not evolve from the Tall type.