Quantification of transient increase of the blood–brain barrier permeability to macromolecules by optimized focused ultrasound combined with microbubbles

Radioimmunotherapy using a radiolabeled monoclonal antibody that targets tumor cells has been shown to be efficient for the treatment of many malignant cancers, with reduced side effects. However, the blood–brain barrier (BBB) inhibits the transport of intravenous antibodies to tumors in the brain. Recent studies have demonstrated that focused ultrasound (FUS) combined with microbubbles (MBs) is a promising method to transiently disrupt the BBB for the drug delivery to the central nervous system. To find the optimal FUS and MBs that can induce reversible increase in the BBB permeability, we employed minimally invasive multiphoton microscopy to quantify the BBB permeability to dextran-155 kDa with similar molecular weight to an antibody by applying different doses of FUS in the presence of MBs with an optimal size and concentration. The cerebral microcirculation was observed through a section of frontoparietal bone thinned with a micro-grinder. About 5 minutes after applying the FUS on the thinned skull in the presence of MBs for 1 minute, TRITC (tetramethylrhodamine isothiocyanate)-dextran-155 kDa in 1% bovine serum albumin in mammalian Ringer’s solution was injected into the cerebral circulation via the ipsilateral carotid artery by a syringe pump. Simultaneously, the temporal images were collected from the brain parenchyma ~100–200 μm below the pia mater. Permeability was determined from the rate of tissue solute accumulation around individual microvessels. After several trials, we found the optimal dose of FUS. At the optimal dose, permeability increased by ~14-fold after 5 minutes post-FUS, and permeability returned to the control level after 25 minutes. FUS without MBs or MBs injected without FUS did not change the permeability. Our method provides an accurate in vivo assessment for the transient BBB permeability change under the treatment of FUS. The optimal FUS dose found for the reversible BBB permeability increase without BBB disruption is reliable and can be applied to future clinical trials

Development of in vitro methods to model the impact of vaginal lactobacilli on Staphylococcus aureus biofilm formation on menstrual cups as well as validation of recommended cleaning directions

IntroductionMenstrual cups (MC) are a reusable feminine hygiene product. A recent publication suggested that Staphylococcus aureus (S. aureus) biofilms can form on MCs which may lead to increased risk of menstrual Toxic Shock Syndrome (mTSS). Additionally, there is concern that buildup of residual menses may contribute to microbial growth and biofilm formation further increasing mTSS risk. Quantitative and qualitative analysis of in vitro tests were utilized to determine if S. aureus biofilm could form on MC in the presence of the keystone species Lactobacillus after 12 h of incubation. The methodology was based on a modification of an anaerobic in vitro method that harnesses the keystone species hypothesis by including a representative of vaginal lactic acid bacteria.MethodsMCs were incubated anaerobically for 12 h in Vaginal Defined Media (VDM) with the two morphologically distinct bacteria, Lactobacillus gasseri (L. gasseri) and S. aureus. Colony Forming Units (CFU) for each organism from the VDM broth and sonicated MC were estimated. In addition, a separate experiment was conducted where S. aureus was grown for 12 h in the absence of L. gasseri. Qualitative analysis for biofilm formation utilized micro-CT (µ-CT) and cryogenic scanning electron microscopy (Cryo-SEM).ResultsSamples collected from the media control had expected growth of both organisms after 12 h of incubation. Samples collected from VDM broth were similar to media control at the end of the 12-h study. Total S. aureus cell density on MC following sonication/rinsing was minimal. Results when using a monoculture of S. aureus demonstrated that there was a significant growth of the organism in the media control and broth as well as the sonicated cups indicating that the presence of L. gasseri was important for controlling growth and adherence of S. aureus. Few rod-shaped bacteria (L. gasseri) and cocci (S. aureus) could be identified on the MCs when grown in a dual species culture inoculum and no biofilm was noted via µ-CT and cryo-SEM. Additionally, efforts to model and understand the validity of the current labeled recommendations for MC cleaning in-between uses are supported.DiscussionThe data support continued safe use of the Tampax® cup when used and maintained as recommended

In-situ high resolution dynamic X-ray microtomographic imaging of olive oil removal in kitchen sponges by squeezing and rinsing

Recent advances in high resolution X-ray tomography (mu CT) technology have enabled in-situ dynamic mu CT imaging (4D-mu CT) of time-dependent processes inside 3D structures, non-destructively and non-invasively. This paper illustrates the application of 4D-mu CT for visualizing the removal of fatty liquids from kitchen sponges made of polyurethane after rinsing (absorption), squeezing (desorption) and cleaning (adding detergents). For the first time, time-dependent imaging of this type of system was established with sufficiently large contrast gradient between water (with/without detergent) and olive oil (model fat) by the application of suitable fat-sensitive X-ray contrast agents. Thus, contrasted olive oil filled sponges were rinsed and squeezed in a unique laboratory loading device with a fluid flow channel designed to fit inside a rotating gantry-based X-ray mu CT system. Results suggest the use of brominated vegetable oil as a preferred contrast agent over magnetite powder for enhancing the attenuation coefficient of olive oil in a multi fluid filled kitchen sponge. The contrast agent (brominated vegetable oil) and olive oil were mixed and subsequently added on to the sponge. There was no disintegration seen in the mixture of contrast agent and olive oil during the cleaning process by detergents. The application of contrast agents also helped in accurately tracking the movement and volume changes of soils in compressed open cell structures. With the in house-built cleaning device, it was quantified that almost 99% of cleaning was possible for contrasted olive oil (brominated vegetable oil with olive oil) dispersed in the sponge. This novel approach allowed for realistic mimicking of the cleaning process and provided closer evaluation of the effectiveness of cleaning by detergents to minimize bacterial growth

Quantification of transient increase of the blood-brain barrier permeability to macromolecules by optimized focused ultrasound combined with microbubbles.

Radioimmunotherapy using a radiolabeled monoclonal antibody that targets tumor cells has been shown to be efficient for the treatment of many malignant cancers, with reduced side effects. However, the blood-brain barrier (BBB) inhibits the transport of intravenous antibodies to tumors in the brain. Recent studies have demonstrated that focused ultrasound (FUS) combined with microbubbles (MBs) is a promising method to transiently disrupt the BBB for the drug delivery to the central nervous system. To find the optimal FUS and MBs that can induce reversible increase in the BBB permeability, we employed minimally invasive multiphoton microscopy to quantify the BBB permeability to dextran-155 kDa with similar molecular weight to an antibody by applying different doses of FUS in the presence of MBs with an optimal size and concentration. The cerebral microcirculation was observed through a section of frontoparietal bone thinned with a micro-grinder. About 5 minutes after applying the FUS on the thinned skull in the presence of MBs for 1 minute, TRITC (tetramethylrhodamine isothiocyanate)-dextran-155 kDa in 1% bovine serum albumin in mammalian Ringer's solution was injected into the cerebral circulation via the ipsilateral carotid artery by a syringe pump. Simultaneously, the temporal images were collected from the brain parenchyma ~100-200 μm below the pia mater. Permeability was determined from the rate of tissue solute accumulation around individual microvessels. After several trials, we found the optimal dose of FUS. At the optimal dose, permeability increased by ~14-fold after 5 minutes post-FUS, and permeability returned to the control level after 25 minutes. FUS without MBs or MBs injected without FUS did not change the permeability. Our method provides an accurate in vivo assessment for the transient BBB permeability change under the treatment of FUS. The optimal FUS dose found for the reversible BBB permeability increase without BBB disruption is reliable and can be applied to future clinical trials

In-situ high resolution dynamic X-ray microtomographic imaging of olive oil removal in kitchen sponges by squeezing and rinsing

Recent advances in high resolution X-ray tomography (mu CT) technology have enabled in-situ dynamic mu CT imaging (4D-mu CT) of time-dependent processes inside 3D structures, non-destructively and non-invasively. This paper illustrates the application of 4D-mu CT for visualizing the removal of fatty liquids from kitchen sponges made of polyurethane after rinsing (absorption), squeezing (desorption) and cleaning (adding detergents). For the first time, time-dependent imaging of this type of system was established with sufficiently large contrast gradient between water (with/without detergent) and olive oil (model fat) by the application of suitable fat-sensitive X-ray contrast agents. Thus, contrasted olive oil filled sponges were rinsed and squeezed in a unique laboratory loading device with a fluid flow channel designed to fit inside a rotating gantry-based X-ray mu CT system. Results suggest the use of brominated vegetable oil as a preferred contrast agent over magnetite powder for enhancing the attenuation coefficient of olive oil in a multi fluid filled kitchen sponge. The contrast agent (brominated vegetable oil) and olive oil were mixed and subsequently added on to the sponge. There was no disintegration seen in the mixture of contrast agent and olive oil during the cleaning process by detergents. The application of contrast agents also helped in accurately tracking the movement and volume changes of soils in compressed open cell structures. With the in house-built cleaning device, it was quantified that almost 99% of cleaning was possible for contrasted olive oil (brominated vegetable oil with olive oil) dispersed in the sponge. This novel approach allowed for realistic mimicking of the cleaning process and provided closer evaluation of the effectiveness of cleaning by detergents to minimize bacterial growth