The cellular and mechanistic study of nisin inhibition of bacillus anthracis spore outgrowth, nisin alteration of infection, and native producer immunity

A critical step during Bacillus anthracis infection is the outgrowth of germinated spores into vegetative bacilli that proliferate and disseminate rapidly within the host. An important challenge exists for developing chemotherapeutic agents that act upon and kill B. anthracis immediately after germination initiation when antibiotic resistance is lost, but prior to the outgrowth into vegetative bacilli, which is accompanied by toxin production. Chemical agents must also function in a manner refractive to the development of antimicrobial resistance. In this thesis we have identified the lantibiotics as a class of chemotherapeutics that are predicted to satisfy these two criteria. The objective of this thesis was to evaluate the efficacy of nisin, a prototypical lantibiotic, in prevention of outgrowth of germinated B. anthracis spores. Like all lantibiotics, nisin is a ribosomally translated peptide that undergoes post-translational modification to form (methyl)lanthionine rings that are critical for antimicrobial activity. Our studies indicate that nisin rapidly inhibits the in vitro outgrowth of germinated B. anthracis Sterne 7702 spores. Although germination initiation was shown to be essential for nisin-dependent antimicrobial activity, nisin did not inhibit or promote germination initiation. Nisin irreversibly killed germinated spores by blocking the establishment of a membrane potential and oxidative metabolism, while not affecting the dissolution of the outer spore structures. The membrane permeability of the spore was increased by nisin, but germinated spores did not undergo full lysis. Nisin was demonstrated to localize to lipid II, which is the penultimate precursor for cell wall biogenesis. This localization suggests two possible independent mechanisms of action, membrane pore formation and inhibition of peptidoglycan synthesis. Structure-activity studies with a truncated form of nisin lacking the two C-terminal (methyl)lanthionine rings and with non-pore forming mutants indicated that membrane disruption is essential for nisin-dependent inhibition of spore outgrowth to prevent membrane potential establishment. Finally, utilizing an in vitro infection model, it was shown that nisin reduced the viability of B. anthracis spores within an infection resulting in increased survival of immune cells while reducing infection-mediated cytokine expression. Fluorescence microscopy indicated that nisin localizes with spores within phagosomes of peritioneal macrophages in germinating conditions. These data demonstrate the effectiveness of nisin, as a model lantibiotic, for preventing spore outgrowth. It is speculated that nisin targeting of lipid II, resulting in membrane perturbations, may be effective at inhibiting the outgrowth of spores prepared from bacteria across a number of species.

Modeling height-diameter relationship in pinus pinaster ait. in the forest intervention zone of Lomba, NE-Portugal

In this work, the relationship between diameter at breast height (d) and total height (h) of individual-tree was modeled with the aim to establish provisory height-diameter (h-d) equations for maritime pine (Pinus pinaster Ait.) stands in the Lomba ZIF, Northeast Portugal. Using data collected locally, several local and generalized h-d equations from the literature were tested and adaptations were also considered. Model fitting was conducted by using usual nonlinear least squares (nls) methods. The best local and generalized models selected, were also tested as mixed models applying a first-order conditional expectation (FOCE) approximation procedure and maximum likelihood methods to estimate fixed and random effects. For the calibration of the mixed models and in order to be consistent with the fitting procedure, the FOCE method was also used to test different sampling designs. The results showed that the local h-d equations with two parameters performed better than the analogous models with three parameters. However a unique set of parameter values for the local model can not be used to all maritime pine stands in Lomba ZIF and thus, a generalized model including covariates from the stand, in addition to d, was necessary to obtain an adequate predictive performance. No evident superiority of the generalized mixed model in comparison to the generalized model with nonlinear least squares parameters estimates was observed. On the other hand, in the case of the local model, the predictive performance greatly improved when random effects were included. The results showed that the mixed model based in the local h-d equation selected is a viable alternative for estimating h if variables from the stand are not available. Moreover, it was observed that it is possible to obtain an adequate calibrated response using only 2 to 5 additional h-d measurements in quantile (or random) trees from the distribution of d in the plot (stand). Balancing sampling effort, accuracy and straightforwardness in practical applications, the generalized model from nls fit is recommended. Examples of applications of the selected generalized equation to the forest management are presented, namely how to use it to complete missing information from forest inventory and also showing how such an equation can be incorporated in a stand-level decision support system that aims to optimize the forest management for the maximization of wood volume production in Lomba ZIF maritime pine stands.

Glucose-dependent insulinotropic polypeptide (GIP) directly affects collagen fibril diameter and collagen cross-linking in osteoblast cultures.

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Relationship between waist circumference and sagittal abdominal diameter measured at different anatomical sites and inflammatory biomarkers in apparently health men

Introduction: Visceral fat accumulation is associated with several changes, such as, increased production of inflammatory biomarkers, especially, C-reactive protein (CRP) and fibrinogen. Anthropometric measurements for central adiposity evaluation, such as, waist circumference (WC) and sagittal abdominal diameter (SAD) have been highlighted. However, there is no consensus on the best anatomical site for measurement. Objective: To evaluate the reliability of different measurements of WC and SAD and verify their capacity to discriminate changes in inflammatory biomarkers. Method: 130 men (20-59 years) were assessed, having measurements of weight, height, WC and SAD. It was considered as the cutoff point for high-sensitivity CRP (hs-CRP) values ≥ 0.12 mg/dL and for fibrinogen the 50th percentile of the evaluated sample. Results: All measurements presented an intraclass correlation coefficient between 0.998 and 0.999. WC measured at the umbilical level (AUC=0.693±0.049) and the smallest circumference between the thorax and the hips (AUC=0.607±0.050) had greater ability to discriminate changes in concentrations of hs-CRP and fibrinogen, respectively. SAD (umbilical level) showed the better ability to detect changes in concentrations of hs-CRP (AUC=0.698± 0.049) and fibrinogen (AUC=0.625±0.049), according to the ROC analysis (p<0.05). Conclusion: WC (smallest circumference between the thorax and the hips) and SAD (umbilical level) are the anatomic sites of measurement for use in predicting the inflammatory risk in apparently health men.

Threshold values of sagittal abdominal diameter for the detection of cardio-metabolic risk factors in Northeastern Mexico: a cross-sectional study

Background: The use of sagittal abdominal diameter (SAD) has been proposed for screening cardio-metabolic risk factors; however, its accuracy can be influenced by the choice of thresholds values. Aim: To determine the SAD threshold values for cardio-metabolic risk factors in Mexican adults; to assess whether parallel and serial SAD testing can improve waist circumference (WC) sensitivity and specificity; and to analyze the effect of considering SAD along with WC and body mass index (BMI) in detecting cardio-metabolic risk. Methods: This cross-sectional study was conducted during 2012-2014 in Northeast Mexico (n = 269). Data on anthropometric, clinical, and biochemical measurements were collected. Sex-adjusted receiver-operating characteristic curves (ROC) were obtained using hypertension, dysglycemia, dyslipidemia and insulin resistance as individual outcomes and metabolic syndrome as a composite outcome. Age-adjusted odds ratios and 95% confidence intervals (CI) were estimated using logistic regression. Results: The threshold value for SAD with acceptable combination of sensitivity and specificity was 24.6 cm in men and 22.5 cm in women. Parallel SAD testing improved WC sensitivity and serial testing improved WC specificity. The co-occurrence of high WC/high SAD increased the risk for insulin resistance by 2.4-fold (95% CI: 1.1-5.3), high BMI/high SAD by 4.3-fold (95% CI: 1.7-11.9) and SAD alone by 2.2-fold (95% CI: 1.2.-4.2). Conclusions: The use of SAD together with traditional obesity indices such as WC and BMI has advantages over using either of these indices alone. SAD may be a powerful screening tool for interventions for high-risk individuals.

Extended studies on fibre diameter measurement with OFDA instrument

OFDA 2000 is a widely used fibre diameter measurement instrument. The instrument can provide improved statistical accuracy measurements in comparison with traditional technology. In this paper, a new technique has been developed to enable fibre diameters to be measured in an aqueous environment using the OFDA 2000 instrument. This new technique was utilised to assess the effects of pH and temperature on the diameters of Merino wool fibres in aqueous environments. Significant changes in fibre diameter under aqueous conditions were found as a function of pH and temperature. It is also demonstrated in this paper that the fibre diameter measurement results can be affected by fibre colour. In both dry and aqueous conditions, variation in fibre diameter was observed on fibres with different colours. This could be due to an optical effect or more likely due to physical changes in the dyed fibres as a result of the presence of dyes. Apart from their physical bulk, dyes may also affect the water content of fibres and hence have an influence on the swelling of fibres when they are measured under ambient regain and wet conditions.

Colour effect on fibre diameter measurement with OFDA 2000

In this study, wool samples from Merino, Corriedale and Lincoln sheep with a wide range of fibre diameters have been dyed and measured for fibre diameter using OFDA 2000 in both dry and conditioned states. Variation in fibre diameter is observed on fibres with different colours in both dry and aqueous conditions. This could be due to an optical effect or more likely due to physical changes in the dyed fibres owing to the presence of dyes. Apart from their physical bulk, dyes may also affect the owing water content of fibres and hence have an influence on the swelling of fibres when they are measured under ambient regain and wet conditions.

Coarser wool is not a necessary consequence of sheep aging: allometric relationship between fibre diameter and fleece-free liveweight of Saxon Merino sheep

The mean fibre diameter (MFD) of wool is the primary determinant of price, processing performance and textile quality. This study determines the primary influences on MFD as Saxon Merino sheep age, by allometrically relating MFD to fleece-free liveweight (FFLwt). In total, 79 sheep were grazed in combinations of three stocking rates and two grazing systems (GS: sheep only; mixed with Angora goats) and studied over 3 years. Measurements were made over 14 consecutive periods (Segments), including segments of FFLwt gain or FFLwt loss. Using shearing and liveweight records and dye-bands on wool, the FFLwt and average daily gain (ADG) of each sheep were determined for each segment. The mean and range in key measurements were as follows: FFLwt, 40.1 (23.1 to 64.1) kg; MFD, 18.8 (12.7 to 25.8) μm. A random coefficient restricted maximum likelihood (REML) regression mixed model was developed to relate the logarithm of MFD to the logarithm of FFLwt and other effects. The model can be written in the form of ${\rm MFD}\,{\equals}\,\rkappa \left( {{\rm GS,}\,{\rm A}{\rm ,}\,{\rm Segment}{\rm .Plot,}\,{\rm Segment,}\,{\rm ADG}} \right){\times}{\rm FFLwt}^{{\left( {\ralpha \left( {{\rm GS}} \right){\plus}\rbeta \left(\rm A \right){\plus}\rgamma \left( {{\rm Segment}{\rm .Plot}} \right)} \right)}} $ , where $\ralpha \left( {{\rm GS}} \right)\,{\equals}\,\;\left\{ {\matrix{\!\! {0.32\left( {{\rm SE}\,{\equals}\,{\rm 0}{\rm .038}} \right)\,{\rm when}\,{\rm sheep}\,{\rm are}\,{\rm grazed}\,{\rm alone}} \hfill \cr \!\!\!\!{0.49\left( {{\rm SE}\,{\equals}\,{\rm 0}{\rm .049}} \right)\,{\rm when}\,{\rm sheep}\,{\rm are}\,{\rm mixed}\,{\rm with}\,{\rm goats}} \hfill \cr } } \right.$ β(A) is a random animal effect, γ(Segment.Plot) a random effect associated with Segment.plot combinations, and κ a constant that depends on GS, random animal effects, random Segment.plot combination effects, Segment and ADG. Thus, MFD was allometrically related to the cube root of FFLwt over seasons and years for sheep, but to the square root of FFLwt for sheep grazed with goats. The result for sheep grazed alone accords with a primary response being that the allocation of nutrients towards the cross-sectional growth of wool follicles is proportional to the changes in the skin surface area arising from changes in the size of the sheep. The proportionality constant varied systematically with ADG, and in sheep only grazing, was about 5 when sheep lost 100 g/day and about 6 when sheep gained 100 g/day. The proportionality constant did not systematically change with chronological age. The variation in the allometric coefficient between individual sheep indicates that some sheep were more sensitive to changes in FFLwt than other sheep. Key practical implications include the following: (a) the reporting of systematic increases in MFD with age is likely to be a consequence of allowing sheep to increase in size during shearing intervals as they age; (b) comparisons of MFD between sheep are more likely to have a biological basis when standardised to a common FFLwt and not just to a common age;

A Model for Continuous Measurement of Drilled Shaft Diameter During Construction

Non-Destructive Testing (NDT) of deep foundations has become an integral part of the industry’s standard manufacturing processes. It is not unusual for the evaluation of the integrity of the concrete to include the measurement of ultrasonic wave speeds. Numerous methods have been proposed that use the propagation speed of ultrasonic waves to check the integrity of concrete for drilled shaft foundations. All such methods evaluate the integrity of the concrete inside the cage and between the access tubes. The integrity of the concrete outside the cage remains to be considered to determine the location of the border between the concrete and the soil in order to obtain the diameter of the drilled shaft. It is also economic to devise a methodology to obtain the diameter of the drilled shaft using the Cross-Hole Sonic Logging system (CSL). Performing such a methodology using the CSL and following the CSL tests is performed and used to check the integrity of the inside concrete, thus allowing the determination of the drilled shaft diameter without having to set up another NDT device. This proposed new method is based on the installation of galvanized tubes outside the shaft across from each inside tube, and performing the CSL test between the inside and outside tubes. From the performed experimental work a model is developed to evaluate the relationship between the thickness of concrete and the ultrasonic wave properties using signal processing. The experimental results show that there is a direct correlation between concrete thicknesses outside the cage and maximum amplitude of the received signal obtained from frequency domain data. This study demonstrates how this new method to measuring the diameter of drilled shafts during construction using a NDT method overcomes the limitations of currently-used methods. In the other part of study, a new method is proposed to visualize and quantify the extent and location of the defects. It is based on a color change in the frequency amplitude of the signal recorded by the receiver probe in the location of defects and it is called Frequency Tomography Analysis (FTA). Time-domain data is transferred to frequency-domain data of the signals propagated between tubes using Fast Fourier Transform (FFT). Then, distribution of the FTA will be evaluated. This method is employed after CSL has determined the high probability of an anomaly in a given area and is applied to improve location accuracy and to further characterize the feature. The technique has a very good resolution and clarifies the exact depth location of any void or defect through the length of the drilled shaft for the voids inside the cage. The last part of study also evaluates the effect of voids inside and outside the reinforcement cage and corrosion in the longitudinal bars on the strength and axial load capacity of drilled shafts. The objective is to quantify the extent of loss in axial strength and stiffness of drilled shafts due to presence of different types of symmetric voids and corrosion throughout their lengths.

Measurement of vein diameter for peripherally inserted central catheter (PICC) insertion: an observational study

Choosing an appropriately sized vein reduces the risk of venous thromboembolism associated with peripherally inserted central catheters. This observational study described the diameters of the brachial, basilic, and cephalic veins and determined the effect of patient factors on vein size. Ultrasound was used to measure the veins of 176 participants. Vein diameter was similar in both arms regardless of hand dominance and side. Patient factors-including greater age, height, and weight, as well as male gender-were associated with increased vein diameter. The basilic vein tended to have the largest diameter statistically. However, this was the case in only 55% of patients.

Vein diameter for peripherally inserted cathether insertion: a scoping review

Background: The risk of venous thromboembolism (VTE) may be reduced if a vein of appropriate diameter is used forperipherally inserted central catheter (PICC) insertion. However, clinicians may have predilections to cannulate certainvein types and use particular insertion sites (eg, right or left arm) and therefore do not necessarily assess all veinsavailable to determine the most optimal vessel to introduce a catheter. It is important that clinicians have anunderstanding of the diameter of veins used for PICC insertion and the effect of patient factors such as hand dominanceon vein size to determine whether their clinical practice is appropriate.

Methods: A scoping review of published literature was performed to determine existing knowledge regarding thediameters of veins used for PICC insertion and the influence of patient factors such as hand dominance and laterality(left or right arm) on vein size.

Results: There was limited published research about the diameters of the basilic, brachial, and cephalic veins at themidupper arm, with only 6 studies identified. Three of the 6 selected articles focused on vein diameter measurement toinform arteriovenous fistula development. Only 1 study included participants undergoing PICC insertion. Scant researchexamined the effect of laterality on vein diameter and 1 study was identified that reported the influence of handdominance or vein type on the diameter of veins used for PICC insertion.

Conclusions: This review found that there is a paucity of studies that have examined the veins used for PICC insertion.Nevertheless, it appears that the basilic vein has the largest diameter (with smaller brachial and cephalic veins),although this is not always the case. Laterality and hand dominance does not seem to influence vein diameter. Furtherresearch about the vasculature used for PICC insertion is needed to inform clinical practice.

Long term performance evaluation of small-diameter vascular grafts based on polyvinyl alcohol hydrogel and dextran and MSCs-based therapies using the ovine pre-clinical animal model.

The functional and structural performance of a 5 cm synthetic small diameter vascular graft (SDVG) produced by the copolymerization of polyvinyl alcohol hydrogel with low molecular weight dextran (PVA/Dx graft) associated to mesenchymal stem cells (MSCs)-based therapies and anticoagulant treatment with heparin, clopidogrel and warfarin was tested using the ovine model during the healing period of 24 weeks. The results were compared to the ones obtained with standard expanded polyetetrafluoroethylene grafts (ePTFE graft). Blood flow, vessel and graft diameter measurements, graft appearance and patency rate (PR), thrombus, stenosis and collateral vessel formation were evaluated by B-mode ultrasound, audio and color flow Doppler. Graft and regenerated vessels morphologic evaluation was performed by scanning electronic microscopy (SEM), histopathological and immunohistochemical analysis. All PVA/Dx grafts could maintain a similar or higher PR and systolic / diastolic laminar blood flow velocities were similar to ePTFE grafts. CD14 (macrophages) and α-actin (smooth muscle) staining presented similar results in PVA/Dx/MSCs and ePTFE graft groups. Fibrosis layer was lower and endothelial cells were only detected at graft-artery transitions where it was added the MSCs. In conclusion, PVA/Dx graft can be an excellent scaffold candidate for vascular reconstruction, including clinic mechanically challenging applications, such as SDVGs, especially when associated to MSCs-based therapies to promote higher endothelialization and lower fibrosis of the vascular prosthesis, but also higher PR values.

Growth and survival of Sclerolobium paniculatum vogel and the relationship between rainfall and the increment in diameter at different planting spacings.

Sclerolobium paniculatum Vogel is a species that has good potential for reclamation of degraded soils. The aim of the investigation was to evaluate the growth and survival of the species and the influence of rainfall on growth in diameter as a function of different spacings (4 m x 2 m, 4 m x 3 m, and 4 m x 4 m). The results indicate that the temporal analysis (period from November 2007 to August 2013) detected significant differences (p ? 0.05) in height between the 4 m x 2 m and 4 m x 4 m spacings, while no significant difference in diameter was found between the 4 m x 2 m and 4 m x 3 m spacings. However, the statistical differences did not persist when the data was analyzed at seven and half years old. Regarding survival, a significant difference was observed only between the 4 m x 4 m spacing and the others, with superiority to the former. A strong correlation was found between rainfall and the increment in diameter of individuals in the broader spacings (R = 0.80 in the 4 m x 3 m spacing and R = 0.77 in the 4 m x 4 m spacing), while in the denser spacing the correlation was moderate (R = 0.56 in the 4 m x 2 m spacing). Since the spacings adopted did not influence tree growth by the end of the period, the choice will depend on other factors such as survival and costs of implementation and forestry management. Plantations in regions with larger rainfall amplitude may benefit the productivity of the species.

Characterization of expiration air jets and droplet size distributions immediately at the mouth opening

Size distributions of expiratory droplets expelled during coughing and speaking and the velocities of the expiration air jets of healthy volunteers were measured. Droplet size was measured using the Interferometric Mie imaging (IMI) technique while the Particle Image Velocimetry (PIV) technique was used for measuring air velocity. These techniques allowed measurements in close proximity to the mouth and avoided air sampling losses. The average expiration air velocity was 11.7 m/s for coughing and 3.9 m/s for speaking. Under the experimental setting, evaporation and condensation effects had negligible impact on the measured droplet size. The geometric mean diameter of droplets from coughing was 13.5m and it was 16.0m for speaking (counting 1 to 100). The estimated total number of droplets expelled ranged from 947 – 2085 per cough and 112 – 6720 for speaking. The estimated droplet concentrations for coughing ranged from 2.4 - 5.2cm-3 per cough and 0.004 – 0.223 cm-3 for speaking.

New particle formation and growth at a remote, sub-tropical coastal location

A month-long intensive measurement campaign was conducted in March/April 2007 at Agnes Water, a remote coastal site just south of the Great Barrier Reef on the east coast of Australia. Particle and ion size distributions were continuously measured during the campaign. Coastal nucleation events were observed in clean, marine air masses coming from the south-east on 65% of the days. The events usually began at ~10:00 local time and lasted for 1-4 hrs. They were characterised by the appearance of a nucleation mode with a peak diameter of ~10 nm. The freshly nucleated particles grew within 1-4 hrs up to sizes of 20-50 nm. The events occurred when solar intensity was high (~1000 W m-2) and RH was low (~60%). Interestingly, the events were not related to tide height. The volatile and hygroscopic properties of freshly nucleated particles (17-22.5 nm), simultaneously measured with a volatility-hygroscopicity-tandem differential mobility analyser (VH-TDMA), were used to infer chemical composition. The majority of the volume of these particles was attributed to internally mixed sulphate and organic components. After ruling out coagulation as a source of significant particle growth, we conclude that the condensation of sulphate and/or organic vapours was most likely responsible for driving particle growth during the nucleation events. We cannot make any direct conclusions regarding the chemical species that participated in the initial particle nucleation. However, we suggest that nucleation may have resulted from the photo-oxidation products of unknown sulphur or organic vapours emitted from the waters of Hervey Bay, or from the formation of DMS-derived sulphate clusters over the open ocean that were activated to observable particles by condensable vapours emitted from the nutrient rich waters around Fraser Island or Hervey Bay. Furthermore, a unique and particularly strong nucleation event was observed during northerly wind. The event began early one morning (08:00) and lasted almost the entire day resulting in the production of a large number of ~80 nm particles (average modal concentration during the event was 3200 cm-3). The Great Barrier Reef was the most likely source of precursor vapours responsible for this event.