The rational design of a synthetic polymer nanoparticle that neutralizes a toxic peptide in vivo

Synthetic polymer nanoparticles (NPs) that bind venomous molecules and neutralize their function in vivo are of significant interest as "plastic antidotes." Recently, procedures to synthesize polymer NPs with affinity for target peptides have been reported. However, the performance of synthetic materials in vivo is a far greater challenge. Particle size, surface charge, and hydrophobicity affect not only the binding affinity and capacity to the target toxin but also the toxicity of NPs and the creation of a "corona" of proteins around NPs that can alter and or suppress the intended performance. Here, we report the design rationale of a plastic antidote for in vivo applications. Optimizing the choice and ratio of functional monomers incorporated in the NP maximized the binding affinity and capacity toward a target peptide. Biocompatibility tests of the NPs in vitro and in vivo revealed the importance of tuning surface charge and hydrophobicity to minimize NP toxicity and prevent aggregation induced by nonspecific interactions with plasma proteins. The toxin neutralization capacity of NPs in vivo showed a strong correlation with binding affinity and capacity in vitro. Furthermore, in vivo imaging experiments established the NPs accelerate clearance of the toxic peptide and eventually accumulate in macrophages in the liver. These results provide a platform to design plastic antidotes and reveal the potential and possible limitations of using synthetic polymer nanoparticles as plastic antidotes.     

Ethylmaltol Odor Enhances Salivary Hemodynamic Responses to Sucrose Taste as Detected by Near-Infrared Spectroscopy

The perceived intensity of the taste of a food is enhanced only if the odor of the food is perceptually similar to the taste. For example, a caramel-like odor enhances the perceived intensity of sweetness. The way gustatory and olfactory signals are integrated in the brain depends largely on one’s previous experiences with taste and odor pairings. To elucidate the effects of a sweet, sugary odor, ethylmaltol, on sucrose taste, as perceived by the central integration of flavor, we recorded salivary hemodynamic responses to the odor and taste pairings using near-infrared spectroscopy (NIRS) of seven panelists. First, we observed concentration-dependent increases in the amplitude of the responses to 0 to 6 % sucrose solutions. Second, when ethylmaltol odor was added to a 4 % sucrose solution, we observed a significant increase in the amplitude of the responses from all panelists. The addition of ethylmaltol to a tasteless solution caused no significant change in the amplitude of the salivary hemodynamic response. These results indicate that the sweet odor of ethylmaltol enhances the salivary hemodynamic response when combined with a sweet taste. Therefore, a congruent combination of sweet odor and taste greatly enhances salivary responses, which are dependent on the central integration of odor and taste.     

New Generation Extracorporeal Membrane Oxygenation With MedTech Mag‐Lev,a Single‐Use,Magnetically Levitated,Centrifugal Blood Pump: Preclinical Evaluation in Calves

We have evaluated the feasibility of a newly developed single‐use, magnetically levitated centrifugal blood pump, MedTech Mag‐Lev, in a 3‐week extracorporeal membrane oxygenation (ECMO) study in calves against a Medtronic Bio‐Pump BPX‐80. A heparin‐ and silicone‐coated polypropylene membrane oxygenator MERA NHP Excelung NSH‐R was employed as an oxygenator. Six healthy male Holstein calves with body weights of about 100 kg were divided into two groups, four in the MedTech group and two in the Bio‐Pump group. Under general anesthesia, the blood pump and oxygenator were inserted extracorporeally between the main pulmonary artery and the descending aorta via a fifth left thoracotomy. Postoperatively, both the pump and oxygen flow rates were controlled at 3 L/min. Heparin was continuously infused to maintain the activated clotting time at 200–240 s. All the MedTech ECMO calves completed the study duration. However, the Bio‐Pump ECMO calves were terminated on postoperative days 7 and 10 because of severe hemolysis and thrombus formation. At the start of the MedTech ECMO, the pressure drop across the oxygenator was about 25 mm Hg with the pump operated at 2800 rpm and delivering 3 L/min flow. The PO₂ of the oxygenator outlet was higher than 400 mm Hg with the PCO₂ below 45 mm Hg. Hemolysis and thrombus were not seen in the MedTech ECMO circuits (plasma‐free hemoglobin [PFH] < 5 mg/dL), while severe hemolysis (PFH > 20 mg/dL) and large thrombus were observed in the Bio‐Pump ECMO circuits. Plasma leakage from the oxygenator did not occur in any ECMO circuits. Three‐week cardiopulmonary support was performed successfully with the MedTech ECMO without circuit exchanges. The MedTech Mag‐Lev could help extend the durability of ECMO circuits by the improved biocompatible performances.     

Strategy to differentiate autoimmune pancreatitis from pancreas cancer

Autoimmune pancreatitis (AIP) is a newly described entity of pancreatitis in which the pathogenesis appears to involve autoimmune mechanisms. Based on histological and immunohistochemical examinations of various organs of AIP patients, AIP appears to be a pancreatic lesion reflecting a systemic "IgG4-related sclerosing disease". Clinically, AIP patients and patients with pancreatic cancer share many features, such as preponderance of elderly males, frequent initial symptom of painless jaundice, development of new-onset diabetes mellitus, and elevated levels of serum tumor markers. It is of uppermost importance not to misdiagnose AIP as pancreatic cancer. Since there is currently no diagnostic serological marker for AIP, and approach to the pancreas for histological examination is generally difficult, AIP is diagnosed using a combination of clinical, serological, morphological, and histopathological features. Findings suggesting AIP rather than pancreatic cancer include: fluctuating obstructive jaundice; elevated serum IgG4 levels; diffuse enlargement of the pancreas; delayed enhancement of the enlarged pancreas and presence of a capsule-like rim on dynamic computed tomography; low apparent diffusion coefficient values on diffusion-weighted magnetic resonance image; irregular narrowing of the main pancreatic duct on endoscopic retrograde cholangiopancreatography; less upstream dilatation of the main pancreatic duct on magnetic resonance cholangiopancreatography, presence of other organ involvement such as bilateral salivary gland swelling, retroperitoneal fibrosis and hilar or intrahepatic sclerosing cholangitis; negative work-up for malignancy including endoscopic ultrasound-guided fine needle aspiration; and steroid responsiveness. Since AIP responds dramatically to steroid therapy, accurate diagnosis of AIP can avoid unnecessary laparotomy or pancreatic resection.     

Six-transmembrane epithelial antigen of prostate4 (STEAP4) is a tumor necrosis factor alpha-induced protein that regulates IL-6, IL-8, and cell proliferation in synovium from patients with rheumatoid arthritis

Human six-transmembrane epithelial antigen of prostate4 (STEAP4), an ortholog of mouse tumor necrosis factor-α-induced adipose-related protein (TIARP), plays a role in tumor necrosis factor (TNF)-dependent arthritis models. However, its role in rheumatoid arthritis (RA) is still obscure. This study explored such a role for STEAP4. The expressions of STEAP4, TNFα, and IL-6 were compared in synovia of RA and osteoarthritis patients. STEAP4 induction was examined in TNFα-stimulated fibroblast-like synoviocytes (FLS) in vitro. FLS (with/without TNFα stimulation) were also analyzed for IL-6 expression after STEAP4 knockdown, using siRNA or transfection with STEAP4-plasmid DNA. IL-8, cell proliferation, and apoptosis were also evaluated in STEAP4-overexpressing FLS. The expression of STEAP4 in joints correlated with TNFα expression, specifically in RA synovium. In the cultured FLS, STEAP4 protein expression was augmented by TNFα activation, and localized in endosomal/lysosomal compartments. STEAP4 downregulation by siRNA enhanced the expression of IL-6 mRNA, while STEAP4 overexpression suppressed IL-6 and IL-8 expression, inhibited cell proliferation, and induced apoptosis via caspase-3. The results indicated that human STEAP4 is regulated by TNFα in synovium, where it controls IL-6 secretion and proliferation of FLS, suggesting that STEAP4 might potentially suppress the pathogenesis of TNFα-induced arthritis such as RA.     

Three-dimensional evaluation of volume change in ossification of the posterior longitudinal ligament of the cervical spine using computed tomography

Background

Two-dimensional imaging is not adequate for evaluating ossification of the posterior longitudinal ligament (OPLL). This study was designed to evaluate the accuracy of a novel computed tomography (CT)-based three-dimensional (3D) analysis method that we had devised to measure volume changes in OPLL.

Subjects and methods

Twenty OPLL patients (12 male and 8 female; mean age 63.6 years) who were being followed conservatively were examined twice with an interval of at least 1 year between the two scans. The mean interval was 22 (range 12–45) months. A 3D model was created with DICOM data from CT images, using the MIMICS^® software to calculate the volume. The mean ossification volume was determined from two measurements. Since ossification size varies widely, evaluation of change in volume is generally affected by the original size. Therefore, the change in ossification volume between the first and second CT examinations was calculated as the annual rate of progression.

Results

The type of OPLL was classified as continuous in 3 patients, segmented in 3, and mixed in 14. The mean ossification volume was 1,831.68 mm³ at the first examination and 1,928.31 mm³ at the second, showing a significant mean increase in ossification volume. The mean annual rate of lesion increase was 3.33 % (range 0.08–7.79 %).

Conclusion

The 3D method used allowed detailed OPLL classification and quantification of change in the ossified volume. Thus, this method appears to be very useful for quantitative evaluation of OPLL with only minimal measurement error.     

In vivo pre- and postoperative three-dimensional knee kinematics in unicompartmental knee arthroplasty

Background

Pre- and postoperative knee kinematics in unicompartmental knee arthroplasty (UKA) can be theoretically related to clinical outcome and longevity after UKA with regard to ligament function and the degree of arthritic changes. However, the preoperative knee kinematics of patients indicated for UKA remain to be elucidated, and it is also unclear whether the preoperative kinematics can be maintained by the UKA procedure. The objective of this study was to examine the in vivo pre- and postoperative three-dimensional knee kinematics in UKA while referencing the normal knee kinematics reported in our previous study.

Methods

We analyzed the knee kinematics in 17 knees (14 patients) undergoing UKA via a three-dimensional to two-dimensional registration technique employing femoral condylar translation and femoral axial rotation. The pre- and postoperative knee kinematics during squat motion were evaluated in the same subjects, employing consistent evaluation parameters.

Results

On average, both pre- and postoperative knee kinematics in the range 10–100° of knee flexion demonstrated near-consistent femoral external rotation and anterior translation of the medial condyle and posterior translation of the lateral condyle. However, the mean femoral external rotation angle and the posterior translation of the lateral condyle postoperatively were significantly smaller than the values observed preoperatively.

Discussion

Although the patterns of preoperative knee motion were similar to those seen in normal knees, the magnitude of this motion varied widely between patients, so it was not necessarily representative of normal knees. These variations may be due to the varying degrees of arthritic changes caused by osteoarthritis. Although the patterns of knee kinematics were largely maintained by the UKA procedure, the causes of the significant reductions in the magnitude of motion upon performing the UKA procedure should be investigated in subsequent studies with a larger number of patients.