Evaluation of a Desktop 3D Printed Rigid Refractive-Indexed-Matched Flow Phantom for PIV Measurements on Cerebral Aneurysms.

PURPOSE: Fabrication of a suitable flow model or phantom is critical to the study of biomedical fluid dynamics using optical flow visualization and measurement methods. The main difficulties arise from the optical properties of the model material, accuracy of the geometry and ease of fabrication. METHODS: Conventionally an investment casting method has been used, but recently advancements in additive manufacturing techniques such as 3D printing have allowed the flow model to be printed directly with minimal post-processing steps. This study presents results of an investigation into the feasibility of fabrication of such models suitable for particle image velocimetry (PIV) using a common 3D printing Stereolithography process and photopolymer resin. RESULTS: An idealised geometry of a cerebral aneurysm was printed to demonstrate its applicability for PIV experimentation. The material was shown to have a refractive index of 1.51, which can be refractive matched with a mixture of de-ionised water with ammonium thiocyanate (NH4SCN). The images were of a quality that after applying common PIV pre-processing techniques and a PIV cross-correlation algorithm, the results produced were consistent within the aneurysm when compared to previous studies. CONCLUSIONS: This study presents an alternative low-cost option for 3D printing of a flow phantom suitable for flow visualization simulations. The use of 3D printed flow phantoms reduces the complexity, time and effort required compared to conventional investment casting methods by removing the necessity of a multi-part process required with investment casting techniques.

PIV Analysis of Stented Haemodynamics in the Descending Aorta.

Cardiovascular diseases (CVD) are the leading cause of death in the developed world and aortic aneurysm is a key contributor. Aortic aneurysms typically occur in the thoracic aorta and can extend into the descending aorta. The Frozen Elephant Trunk stent (FET) is one of the leading treatments for the aneurysms extending into the descending aorta. This study focuses on the in-vitro experimentation of a stented descending aorta, investigating the haemodynamics in a compliant phantom. A silicone phantom of the descending aorta was manufactured using a lost core casting method. A PVC stent was manufactured using the same mould core. Particle Image Velocimetry (PIV) was used for pulsatile studies, focusing specifically on the passive fixation at the distal end of the FET. The results showed an apparent expansion in the diastolic period that was identified to be a collapse in the lateral plane. Flow recirculation regions were identified during the collapse. The collapse was attributed to low upstream and high downstream pressures causing a vacuum effect. The findings may imply a potential risk introduced by the FET stent that requires further investigation.

A Review of Arterial Phantom Fabrication Methods for Flow Measurement Using PIV Techniques.

Cardiovascular diseases (CVD) are the leading cause of morbidity and mortality in the western world. In the last three decades, fluid dynamics investigations have been an important component in the study of the cardiovascular system and CVD. A large proportion of studies have been restricted to computational fluid dynamic (CFD) modeling of blood flow. However, with the development of flow measurement techniques such as particle image velocimetry (PIV), and recent advances in additive manufacturing, experimental investigation of such flow systems has become of interest to validate CFD studies, testing vascular implants and using the data for therapeutic procedures. This article reviews the technical aspects of in-vitro arterial flow measurement with the focus on PIV. CAD modeling of geometries and rapid prototyping of molds has been reviewed. Different processes of casting rigid and compliant models for experimental analysis have been reviewed and the accuracy of construction of each method has been compared. A review of refractive index matching and blood mimicking flow circuits is also provided. Methodologies and results of the most influential experimental studies are compared to elucidate the benefits, accuracy and limitations of each method.

Experimental measurement of breath exit velocity and expirated bloodstain patterns produced under different exhalation mechanisms.

In an attempt to obtain a deeper understanding of the factors which determine the characteristics of expirated bloodstain patterns, the mechanism of formation of airborne droplets was studied. Hot wire anemometry measured air velocity, 25 mm from the lips, for 31 individuals spitting, coughing and blowing. Expirated stains were produced by the same mechanisms performed by one individual with different volumes of a synthetic blood substitute in their mouth. The atomization of the liquid at the lips was captured with high-speed video, and the resulting stain patterns were captured on paper targets. Peak air velocities varied for blowing (6 to 64 m/s), spitting (1 to 64 m/s) and coughing (1 to 47 m/s), with mean values of 12 m/s (blowing), 7 m/s (spitting) and 4 m/s (coughing). There was a large (55-65%) variation between individuals in air velocity produced, as well as variation between trials for a single individual (25-35%). Spitting and blowing involved similar lip shapes. Blowing had a longer duration of airflow, though it is not the duration but the peak velocity at the beginning of the air motion which appears to control the atomization of blood in the mouth and thus stain formation. Spitting could project quantities of drops at least 1600 mm. Coughing had a shorter range of near 500 mm, with a few droplets travelling further. All mechanisms could spread drops over an angle >45°. Spitting was the most effective for projecting drops of blood from the mouth, due to its combination of chest motion and mouth shape producing strong air velocities. No unique method was found of inferring the physical action (spitting, coughing or blowing) from characteristics of the pattern, except possibly distance travelled. Diameter range in expirated bloodstains varied from very small (<1 mm) in a dense formation to several millimetres. No unique method was found of discriminating expirated patterns from gunshot or impact patterns on stain shape alone. Only 20% of the expirated patterns produced in this study contained identifiable bubble rings or beaded stains.

Serum fibroblastic growth factor 23 in acute Sarcoidosis and normal kidney function.

BACKGROUND: Serum fibroblastic growth factor (FGF) 23 has recently been established as a major physiological regulator of phosphate homeostasis and may have a causal role in adverse cardiovascular and bone outcomes. However its role in states of disordered phosphate homeostasis and normal kidney function is as yet under characterised. AIMS: To investigate whether this biomarker of vascular calcification and adverse bone outcomes is detectable in patients with sarcoidosis. DESIGN: We conducted a cross sectional study on a convenience sample of patients presenting with acute sarcoidosis to a respiratory tertiary referral unit. METHODS: We set out to systematically examine the characteristics and determinants of serum FGF-23 in patients presenting with acute sarcoidosis. RESULTS: We studied 39 patients, 26 were male. Mean (SD) age was 33 (9.6) years. 15.4% of patients had a serum level of FGF-23 ≥ 9.9 pg/mL. The remaining 84.6% of patients had a serum FGF-23 < 9.9 pg/mL. Those with a detectable serum FGF-23 had a significantly higher serum calcium (P = 0.007), and lower serum iPTH (P<0.001). Serum phosphate and 25-hydroxyvitamin D were not statistically significantly different between groups (P=0.25 and P=0.83). The proportion of patients with stage II disease on CXR was higher in those with a detectable FGF-23 (P<0.001). CONCLUSIONS: Serum FGF-23 was below the level of detection in the majority of this cohort of patients presenting with acute sarcoidosis. A detectable serum FGF-23 was associated with a higher serum calcium and lower serum iPTH.

An Experimental and Numerical Investigation of CO2 Distribution in the Upper Airways During Nasal High Flow Therapy.

Nasal high flow (NHF) therapy is used to treat a variety of respiratory disorders to improve patient oxygenation. A CO2 washout mechanism is believed to be responsible for the observed increase in oxygenation. In this study, experimentally validated Computational Fluid Dynamics simulations of the CO2 concentration within the upper airway during unassisted and NHF assisted breathing were undertaken with the aim of exploring the existence of this washout mechanism. An anatomically accurate nasal cavity model was generated from a CT scan and breathing was reproduced using a Fourier decomposition of a physiologically measured breath waveform. Time dependent CO2 profiles were obtained at the entrance of the trachea in the experimental model, and were used as simulation boundary conditions. Flow recirculation features were observed in the anterior portion of the nasal cavity upon application of the therapy. This causes the CO2 rich gas to vent from the nostrils reducing the CO2 concentration in the dead space and lowering the inspired CO2 volume. Increasing therapy flow rate increases the penetration depth within the nasal cavity of the low CO2 concentration gas. A 65% decrease in inspired CO2 was observed for therapy flow rates ranging from 0 to 60 L min(-1) supporting the washout mechanism theory.

Experimental and computational investigation of the trajectories of blood drops ejected from the nose.

Blood expirated from the nose may leave a characteristic bloodstain at a crime scene which can provide important clues for reconstructing events during a violent assault. Little research has been done on the typical velocities, trajectories and size distribution that can be expected from expirated blood. An experimental fluid dynamics technique known as stereoscopic particle image velocimetry is used in this work to obtain the air velocity field inside and outside the nostrils during exhalation. A numerical model was then used to compute the trajectory of blood drops of 0.5 and 2 mm. The drops were tracked until ground plane impact below the nostril exit. Three heights were investigated, 1.5, 1.6 and 1.7 m. For an expiration flow rate of 32 l/min in vivo, there is a maximum exit velocity from the nostril of approximately 4 m/s, with a 0.5 m/s difference between nostrils. After the drops have traversed the distances investigated, drops of 0.5 and 2 mm in diameter from both nostrils are at a similar velocity. This implies that the gravitational acceleration after the drops leave the jet has the most influence on velocity. It is however shown that exit velocity does affect impact location. Drop size affects both impact location and impact velocity. An increase in height increases the distance traversed. Compared to the 2-mm drop, the 0.5 mm had a lower impact velocity, but its impact location in the ground plane was further from the nostril exit. Understanding the physics of expirated blood flight allows better interpretation of expirated stains at crime scenes.

Visualization of the air ejected from the temporary cavity in brain and tissue simulants during gunshot wounding.

One hypothesis for the physical mechanism responsible for backspatter during cranial gunshot wounding is that air is ejected by the collapse of the temporary cavity formed around the bullet path. Using bovine and ovine heads and simulant materials, evidence of this ejection was sought by measuring the velocity of the air that was drawn in and ejected from the cavity in front of the wound channel after bullet impact. A laminar flow of fog-laden air was arranged in front of the wound channel and two high speed cameras recording at 30,000 frames/second captured the air motion. All samples were shot with standard 9 mm × 19 mm FMJ ammunition. Different concentrations of ballistic gelatine were used to characterize the effect of elasticity of the material on the velocity of the air. Fresh bovine and ovine heads were shot with the same experimental set up to investigate if there was induction of air into, and ejection of air from the entrance wounds. The results show, for the first time, that the temporary cavity does eject air in gelatine. The velocity of in-drawn air for 3, 5 and 10% concentration of gelatine was 81, 76 and 65 m/s respectively and the velocity of ejected air for 5 and 10% concentration of gelatine were 43 and 72 m/s respectively. The results show that when the concentration of gelatine is increased, the velocity of the air drawn into the cavity decreases and the velocity of the ejected air increases. However, no ejection was observed in 3% gelatine, ovine or bovine heads. Although ejection of air was not observed, ejection of brain from the wound channel was seen. Using the velocity of the ejected brain, the minimum intracranial pressure required to eject the brain tissue was estimated to be 712 kPa and 468 kPa for the sheep and bovine heads respectively.

Antibodies anti-Shiga toxin 2 B subunit from chicken egg yolk: isolation, purification and neutralization efficacy.

Shiga toxins (Stx1 and Stx2) are the main virulence factors of enterohemorrhagic Escherichia coli (EHEC), a foodborne pathogen associated with diarrhea, hemorrhagic colitis and hemolytic uremic syndrome. The aim of this study was to evaluate the antibodies against Stx2 obtained from egg yolks of laying hens immunized with a recombinant Stx2B subunit. A high specific response in serum was observed 25 days after the first immunization and IgY antibodies were extracted from day 47th and purified from egg yolk. A concentration of 0.84 mg of total IgY/ml of egg yolk was obtained, of which 8% were antigen specific. The ability of anti-Stx2B IgY to recognize Stx2B and Stx2 either in solid-phase or in solution were evaluated and compared with anti-Stx2B rabbit antibodies by Western blotting and ELISA. The protective efficacy of IgY against Stx2 was determined by in vitro and in vivo experiments. The results show that IgY was able to recognize Stx2B and Stx2 in denatured conditions, attached to a solid-phase and free in solution. The anti-Stx2B IgY could effectively block the biological activity of Stx2 on Vero cells and protect mice from Stx2 challenge. The data suggest that immunization of hens with Stx2B could be a strategy to obtain at low cost a relatively high concentration of anti-Stx2 egg yolk IgY, able to neutralize Stx2 lethal activity. IgY technology could be an useful tool for research, diagnosis and therapy of EHEC infection.

Preclinical assessment of the neutralizing capacity of antivenoms produced in six Latin American countries against medically-relevant Bothrops snake venoms.

Species of the genus Bothrops induce the vast majority of snakebite envenomings in Latin America. A preclinical study was performed in the context of a regional network of public laboratories involved in the production, quality control and development of antivenoms in Latin America. The ability of seven polyspecific antivenoms, produced in Argentina, Brazil, Peru, Bolivia, Colombia and Costa Rica, to neutralize lethal, hemorrhagic, coagulant, defibrinogenating and myotoxic activities of the venoms of Bothrops neuwiedi (diporus) (Argentina), Bothrops jararaca (Brazil), B. neuwiedi (mattogrossensis) (Bolivia), Bothrops atrox (Peru and Colombia) and Bothrops asper (Costa Rica) was assessed using standard laboratory tests. Despite differences in the venom mixtures used in the immunization of animals for the production of these antivenoms, a pattern of extensive cross-neutralization was observed between these antivenoms and all the venoms tested, with quantitative differences in the values of effective doses. This study reveals the capacity of these antivenoms to neutralize, in preclinical tests, homologous and heterologous Bothrops venoms in Central and South America, and also highlight quantitative differences in the values of Median Effective Doses (ED50s) between the various antivenoms.

Characterization of a basic phospholipase A2-homologeu myotoxin isolated from the venom of the snake Bothrops neuwiedii (yarará chica) from Argentina.

A basic protein was isolated by CM-Sephadex C-25 chromatography from the venom of Bothrops neuwiedii from Argentina, and named B. neuwiedii myotoxin I. This protein exerted local myotoxic and edema-forming effects in mice, with potencies comparable to other myotoxins isolated from Bothrops spp. venoms. When injected by i.v. route at doses up to 4.7 mg/kg of body weight, the toxin was not lethal. In vitro, the toxin had no detectable phospholipase A2 activity on egg yolk phospholipids. B. neuwiedii myotoxin I appeared as a homodimer in sodium dodecylsulphate-polyacrylamide gel electrophoresis, with a subunit molecular weight of 15 kD. Gel immunodiffusion revealed a pattern of partial antigenic identity between the newly isolated myotoxin and myotoxin II from Bothrops asper venom. The sequence of B. neuwiedii myotoxin I was determined for the first 40 amino acid residues, showing high homology to several class II phospholipase A2 myotoxins of the Lys-49 family from crotalids. Altogether, results suggest that this toxin is a new member of the Lys-49 phospholipase A2-homologues with myotoxic, cytolytic, and edema-inducing activities.

Comparative study of the cytolytic activity of myotoxic phospholipases A2 on mouse endothelial (tEnd) and skeletal muscle (C2C12) cells in vitro.

A rapid in vitro cytolytic effect of some myotoxic phospholipases A2 (PLA2s) isolated from the venoms of Viperidae snakes has been previously described. This study was undertaken to investigate if cytolytic activity is a common property of the myotoxic proteins from this group. Murine endothelial cells (tEnd) and skeletal muscle myotubes (C2C12) were utilized as targets. The release of lactic dehydrogenase was quantified as a measure of cell damage, 3 h after exposure of cells to the different PLA2s, including representatives from the genera Bothrops, Agkistrodon, Trimeresurus, Crotalus (family Viperidae), and Notechis (family Elapidae). All of the group II myotoxic PLA2s tested displayed rapid cytolytic activity when tested in the micromolar range of concentrations (8-32 microM). In contrast, the group I myotoxic PLA2 notexin was devoid of this activity. Aspartate-49 and lysine-49 PLA2 group II variants showed a comparable cytolytic effect. Skeletal muscle myotubes, obtained after fusion and differentiation of C2C12 myoblasts, were significantly more susceptible to the cytolytic action of myotoxins than endothelial cells, previously reported to be more susceptible than undifferentiated myoblasts under the same assay conditions. Cytolytic activity appears to be a common characteristic of group II myotoxic PLA2s of the Viperidae. Bee venom PLA2, a group III enzyme of known myotoxicity, also displayed cytotoxic activity on C2C12 myotubes, being devoid of activity on endothelial cells. These results suggest that in vitro differentiated skeletal muscle myotubes may represent a suitable model target for the study of myotoxic PLA2s of the structural group II found in snake venoms.

Background levels of bromide in human blood.

A total of 183 random, whole-blood specimens was collected from healthy individuals within the State of Queensland (Australia), and the bromide concentration was determined by wavelength-dispersive X-ray fluorescence spectrometry (WDXRF). The intensity of the tube Compton scatter line was used to account for differences in matrices between the aqueous calibration standard and whole-blood specimens. Technical details of the WDXRF method are included in the experimental section of the paper. The overall mean for bromide in human blood was 5.3 +/- 1.4 mg/L and ranged from 2.5 to 11.7 mg/L. Associations between bromide levels and variables including age, gender, weight, height, and postcode address were examined by ANOVA and Pearson's correlation. Data indicate that aged persons (45-65+ years) are more likely to have higher bromide levels than younger persons (15-25 years). Our results also suggest differences in bromide levels between the sexes in similar age groups. Average levels were higher in females in most age groups. The reason for this difference requires further detailed investigation. No correlation was observed between bromide levels and height or weight of donors. No significant differences in bromide levels were found in persons living in the highly populated southeast region of Queensland compared with those living in the less urbanized northern parts of the state.

Non-organic hearing loss in young persons: transient episode or indicator of deep-seated difficulty.

Depressed auditory thresholds without evidence of organic hearing loss are observed in a small percentage of children and young adults. Several studies suggest that non-organic hearing loss (NOHL) is a manifestation of, or reaction to stress. Proposed treatments range from ignoring the apparent hearing loss through the use of psychotherapy and/or counselling to confronting the individual with evidence to prove that there is no real loss of function. Few studies report on long-term outcome. Thirty-eight subjects who had been diagnosed more than six and up to twenty-seven years ago as having NOHL were identified from records. Efforts were made to determine if there was any evidence of longer term effects. Nine were untraceable. Of those reviewed one was established as a malingerer; five had concurrent speech problems; one was possibly dyslexic; five were or had been under psychiatric care. These findings suggest that NOHL may, in some individuals, indicate underlying problems that merit detailed investigation and treatment.