Carnosine Protects against Cerebral Ischemic Injury by Inhibiting Matrix-Metalloproteinases.

Stroke is one of the leading causes of death and disability worldwide. However, treatment options for ischemic stroke remain limited. Matrix-metalloproteinases (MMPs) contribute to brain damage during ischemic strokes by disrupting the blood-brain barrier (BBB) and causing brain edemas. Carnosine, an endogenous dipeptide, was found by us and others to be protective against ischemic brain injury. In this study, we investigated whether carnosine influences MMP activity. Brain MMP levels and activity were measured by gelatin zymography after permanent occlusion of the middle cerebral artery (pMCAO) in rats and in vitro enzyme assays. Carnosine significantly reduced infarct volume and edema. Gelatin zymography and in vitro enzyme assays showed that carnosine inhibited brain MMPs. We showed that carnosine inhibited both MMP-2 and MMP-9 activity by chelating zinc. Carnosine also reduced the ischemia-mediated degradation of the tight junction proteins that comprise the BBB. In summary, our findings show that carnosine inhibits MMP activity by chelating zinc, an essential MMP co-factor, resulting in the reduction of edema and brain injury. We believe that our findings shed new light on the neuroprotective mechanism of carnosine against ischemic brain damage.

The urethra of healthy female dogs can be normally narrowed due to the urethral flexure in retrograde CT urethrography.

Quantitative analysis of urethral size in male dogs by retrograde CT urethrography using a power injector is a recently reported technique. The aim of the current, prospective, analytical study was to apply the same method to six, healthy, intact female Beagle dogs. The proximal (P < .0001) and middle (P = .0010) urethral volumes significantly differed between the empty and distended bladder states, although no significant difference was observed in the distal volume (P = .0971). Unlike male dogs, female dogs showed two patterns of the urethral course: a straight urethra and urethral flexure. The urethral flexure was always related to intrapelvic position of the urinary bladder, and the more caudal the location of the vesicourethral junction beyond the pecten of the pubic bone, the more marked the appearance of the urethral flexure. Analysis of the urethral diameter at five sites (adopted from the previous radiographic study) was performed, and the urethral diameter showed a significant difference between the empty and the distended bladder states at sites 1 (vesicourethral junction, P < .0001), 2 (P < .0001), and 3 (P = .0244). However, there were no significant differences at sites 4 (P = .2516) and 5 (inflatable retention bulb, P = .1260). The urinary bladder may be in part intrapelvically located in clinically healthy female dogs, and urethral flexure and urethral narrowing can result from the intrapelvic location of the bladder. Narrowing of the urethra should be interpreted with caution when the pelvic bladder is identified.

Retrograde CT urethrography using a power injector quantitatively reveals effects of bladder distension on urethral size in healthy male Beagle dogs.

Quantitative analysis of the normal retrograde urethrogram is well reported in radiography, but studies on CT urethrography are lacking. Recently, a method of retrograde CT urethrography using a power injector was described. The purpose of the current, prospective, analytical study was to quantify the urethral size of five, healthy, intact, male Beagle dogs using retrograde CT urethrography and a power injector. With the injection rate of the power injector set at 0.3 mL/s, 1 mL/kg of diluted contrast medium (15 mg I/mL) was injected, and a CT examination was performed. The state of the initial urethrogram taken was defined as "empty bladder." The same procedures were repeated with the injection of an additional 1 mL/kg of diluted contrast medium until the ureteral reflux was seen (distended bladder). There was a significant difference in volumes between the empty and distended bladder, but the membranous urethra showed the least difference (P = .0044) among the three regions (P < .0001 for the prostatic and penile urethra). Urethral diameters at six sites were measured from sagittal images, and the sites of measurements were adopted from the earlier radiographic studies. The most significant difference in the urethral diameters between the empty and distended bladder occurred at the cranial and middle prostatic urethra (P < .0001). The results of this study can be useful for interpreting the results of retrograde CT urethrography. Care must be taken when narrowing is suspected at the prostatic urethra, and if necessary, further distension of the urinary bladder should be tried.

Methylglyoxal induced advanced glycation end products (AGE)/receptor for AGE (RAGE)-mediated angiogenic impairment in bone marrow-derived endothelial progenitor cells.

Endothelial cells (ECs) maintain the structure and function of blood vessels and are readily exposed to exogenous and endogenous toxic substances in the circulatory system. Bone marrow-derived endothelial progenitor cells (EPCs) circulate in the blood and differentiate to EC, which are known to participate in angiogenesis and regeneration of injured vessels. Dysfunction in EPC contributes to cardiovascular complications in patients with diabetes, but the precise molecular mechanisms underlying diabetic EPC abnormalities are not completely understood. The aim of this study was to investigate the mechanisms underlying diabetic EPC dysfunction using methylglyoxal (MG), an endogenous toxic diabetic metabolite. Data demonstrated that MG decreased cell viability and protein expression of vascular endothelial growth factor receptor (VEGFR)-2 associated with functional impairment of tube formation in EPC. The generation of advanced glycation end (AGE) products was increased in EPC following exposure to MG. Blockage of receptor for AGE (RAGE) by FPS-ZM1, a specific antagonist for RAGE, significantly reversed the decrease of VEGFR-2 protein expression and angiogenic dysfunction in MG-incubated EPC. Taken together, data demonstrated that MG induced angiogenic impairment in EPC via alterations in the AGE/RAGE-VEGFR-2 pathway which may be utilized in the development of potential therapeutic and preventive targets for diabetic vascular complications.

A newly synthesized macakurzin C-derivative attenuates acute and chronic skin inflammation: The Nrf2/heme oxygenase signaling as a potential target.

Impaired immune responses in skin play a pivotal role in the development and progression of chemical-associated inflammatory skin disorders. In this study, we synthesized new flavonoid derivatives from macakurzin C, and identified in vitro and in vivo efficacy of a potent anti-inflammatory flavonoid, Compound 14 (CPD 14), with its underlying mechanisms. In lipopolysaccharide (LPS)-stimulated murine macrophages and IFN-γ/TNF-α-stimulated human keratinocytes, CPD 14 significantly inhibited the release of inflammatory mediators including nitric oxide (NO), prostaglandins, and cytokines (IC50 for NO inhibition in macrophages: 4.61µM). Attenuated NF-κB signaling and activated Nrf2/HO-1 pathway were responsible for the anti-inflammatory effects of CPD 14. The in vivo relevance was examined in phorbol 12-myristate 13-acetate (TPA)-induced acute skin inflammation and oxazolone-induced atopic dermatitis models. Topically applied CPD 14 significantly protected both irritation- and sensitization-associated skin inflammation by suppressing the expression of inflammatory mediators. In summary, we demonstrated that a newly synthesized flavonoid, CPD 14, has potent inhibitory effects on skin inflammation, suggesting it is a potential therapeutic candidate to treat skin disorders associated with excessive inflammation.

Potent Anti-inflammatory and Analgesic Actions of the Chloroform Extract of Dendropanax morbifera Mediated by the Nrf2/HO-1 Pathway.

Dendropanax morbifera LEVEILLE (DP) has been used in traditional Korean medicines to treat a variety of inflammatory diseases. Although the in vitro anti-inflammatory potential of this plant is understood, its in vivo efficacy and underlying molecular mechanism of anti-inflammatory effects are largely unknown. We elucidated the anti-inflammatory and analgesic activities and the underlying molecular mechanisms of DP using in vitro and in vivo models. Lipopolysaccharide (LPS)-stimulated murine macrophages were used to analyze the in vitro anti-inflammatory potential of DP extract and to elucidate the underlying mechanisms. In vivo animal models of phorbol 12-myristate 13-acetate (TPA)-induced ear edema and acetic acid-induced writhing response tests were used to analyze the in vivo anti-inflammatory effects and anti-nociceptive effects of DP extract, respectively. Methanolic extract of DP (DPME) significantly inhibited the release of nitric oxide (NO) and prostaglandin E2 (PGE2) in LPS-activated macrophages. Among the five sub-fractions, the chloroform fraction (DP-C) showed the most potent suppressive effects against pro-inflammatory mediators and cytokines in LPS-stimulated macrophages. These effects were attributed to inhibition of nuclear factor-κB (NF-κB) nuclear translocation and c-Jun N terminal kinase (JNK) 1/2 phosphorylation and to activation of NF-E2-related factor 2/heme oxygenase-1 (Nrf2/HO-1) signaling. DP-C exhibited strong protective in vivo effects in TPA-induced ear edema mouse model and acetic acid-induced writhing response test. Our data suggest that DP-C has potent anti-inflammatory and analgesic activities and may be a promising treatment against a variety of inflammatory diseases.

Protective activity of Dendropanax morbifera against cisplatin-induced acute kidney injury.

BACKGROUND/AIMS: Drug-induced acute kidney injury (AKI) has been a severe threat to hospitalized patients, raising the urgent needs to develop strategies to reduce AKI. We investigated the protective activity of Dendropanax morbifera (DP), a medicinal plant which has been widely used to treat infectious and pain diseases, on acute kidney injury (AKI) using cisplatin-induced nephropathic models. METHODS: Both in vitro renal tubular cells (NRK-52E) and in vivo rat models were used to demonstrate the nephroprotective effect of DP. RESULTS: Methanolic extract from DP significantly reduced cisplatin-induced toxicity in renal tubular cells. Through successive liquid extraction, the extract of DP was separated into n-hexane, CHCl3, EtOAc, n-BuOH, and H2O fractions. Among these, the CHCl3 fraction (DPCF) was found to be most potent. The protective activity of DPCF was found to be mediated through anti-oxidant, mitochondrial protective, and anti-apoptotic activities. In in vivo rat models of AKI, treatment with DPCF significantly reversed the cisplatin-induced increase in blood urea nitrogen and serum creatinine and histopathologic damage, recovered the level of anti-oxidant enzymes, and inhibited renal apoptosis. CONCLUSION: We demonstrated that DP extracts decreased cisplatin-induced renal toxicity, indicating its potential to ameliorate drug-associated acute kidney damage.

Modulation of mitochondrial function and autophagy mediates carnosine neuroprotection against ischemic brain damage.

BACKGROUND AND PURPOSE: Despite the rapidly increasing global burden of ischemic stroke, no therapeutic options for neuroprotection against stroke currently exist. Recent studies have shown that autophagy plays a key role in ischemic neuronal death, and treatments that target autophagy may represent a novel strategy in neuroprotection. We investigated whether autophagy is regulated by carnosine, an endogenous pleiotropic dipeptide that has robust neuroprotective activity against ischemic brain damage. METHODS: We examined the effect of carnosine on mitochondrial dysfunction and autophagic processes in rat focal ischemia and in neuronal cultures. RESULTS: Autophagic pathways such as reduction of phosphorylated mammalian target of rapamycin (mTOR)/p70S6K and the conversion of microtubule-associated protein 1 light chain 3 (LC3)-I to LC3-II were enhanced in the ischemic brain. However, treatment with carnosine significantly attenuated autophagic signaling in the ischemic brain, with improvement of brain mitochondrial function and mitophagy signaling. The protective effect of carnosine against autophagy was also confirmed in primary cortical neurons. CONCLUSIONS: Taken together, our data suggest that the neuroprotective effect of carnosine is at least partially mediated by mitochondrial protection and attenuation of deleterious autophagic processes. Our findings shed new light on the mechanistic pathways that this exciting neuroprotective agent influences.

High glucose condition induces autophagy in endothelial progenitor cells contributing to angiogenic impairment.

Cardiovascular complications are the major causes of death in patients with diabetes mellitus. Several studies have demonstrated that endothelial progenitor cells (EPCs), adult stem cells contributing to the regeneration of vascular endothelium, are dysfunctional under diabetic condition resulting in impaired peripheral circulation and delayed wound healing. In this study, we investigated the cellular alteration of EPCs under high glucose condition, to elucidate the mechanisms underlying diabetes-associated EPC dysfunction. EPCs were isolated from bone marrow and cultured in normal glucose (5.5 mM)- or high glucose (HG; 30 mM)-containing medium. High glucose treated-EPCs showed decreased ability to form tubule-like networks in Matrigel compared to EPCs under normal glucose, which matched well to the clinical observation of diabetic EPC dysfunction. Conversion of LC3-I to LC3-II was increased in EPCs under HG condition, showing that HG induced autophagy in EPCs. Flow cytometric analysis revealed generation of oxidative stress and disruption of mitochondrial permeability in HG exposed EPCs. Increased mitochondrial oxidative stress was also observed by mitochondria-specific superoxide indicator, MitoSOX(TM). Taken together, we demonstrated that autophagy and mitochondrial impairment were induced in EPCs under high glucose condition, giving a new insight into the mechanism underlying dysfunction of diabetic EPCs. We hope that our finding can contribute to the development of a new treatment option for cardiovascular complications in diabetic patients.

Synthetic Strategies for Improving Solubility: Optimization of Novel Pyrazolo[1,5-a]pyrimidine CFTR Activator That Ameliorates Dry Eye Disease.

Dry eye disease (DED) is one of the most prevalent ocular diseases but has limited treatment options. Cystic fibrosis transmembrane conductance regulator (CFTR), a major chloride channel that stimulates fluid secretion in the ocular surface, may pave the way for new therapeutic strategies for DED. Herein, we report the optimization of Cact-3, a potent CFTR activator with poor solubility, to 16d, a potent CFTR activator with suitable solubility for eye drop formulation. Notably, 16d was well distributed in target tissues including cornea and conjunctiva with minimal systemic exposure in rabbit. Topical ocular instillation of 16d significantly enhanced tear secretion and improved corneal erosion in a mouse model of DED. In addition, 16d significantly reduced mRNA expression of pro-inflammatory cytokines including IL-1ß, IL-17, and TNF-α and MMP2 in cornea and conjunctiva of DED mice.

Improved Productivity of Naringin Oleate with Flavonoid and Fatty Acid by Efficient Enzymatic Esterification.

Naringin is a flavonoid found in citrus fruits. It exhibits biological activities, such as anticancer and antioxidant effects, but it suffers from low solubility and low stability in lipophilic systems. These drawbacks lead to difficulties in the commercial application of naringin, but they can be overcome through esterification. In this study, naringin oleate was synthesized by enzymatic esterification and optimal conditions for the reaction were investigated. Experiments were conducted focusing on the following parameters: enzyme type, enzyme concentration, molar ratio of naringin to oleic acid, reaction temperature, and reaction solvent. We further confirmed the degree of esterification based on the difference in the initial and the final naringin concentrations. A conversion of 93.10% was obtained under optimized conditions (Lipozyme TL IM 10 g/L, molar ratio 1:20, reaction temperature 40 °C, acetonitrile as solvent, and 48 h reaction time). Thus, naringin oleate, a high value-added material that overcomes the low hydrophobicity of naringin and enhances its performance, was obtained through esterification of naringin using oleic acid. This study presented a method for the efficient enzymatic synthesis that could ensure high conversion within a shorter reaction time compared with that required in previously reported methods.

Translocation of the conus medullaris during dynamic lumbosacral magnetic resonance imaging in dogs.

OBJECTIVE: To investigate the change in the lumbosacral angle (ΔLSA) and conus medullaris (CM) displacement in healthy dogs undergoing dynamic MRI with changes in the posture of their pelvic limbs from neutral posture to flexion or extension posture and to evaluate for potential correlation between ΔLSA and CM displacement. ANIMALS: 9 healthy adult Beagles. PROCEDURES: Dogs underwent dynamic MRI with their pelvic limbs positioned in neutral, flexion, and extension postures. From T2-weighted sagittal midline plane MRI images, 2 observers measured the lumbosacral angle and CM location in duplicate for each posture for each dog. Intra- and interobserver agreement was assessed, and the Spearman rank correlation coefficient (ρ) was used to assess for potential correlation between ΔLSA and CM displacement for changes in pelvic limb posture from neutral to flexion or extension. RESULTS: Overall, the mean ΔLSA and CM displacement for changes in posture were 23° and 9.09 mm (caudal displacement) for the change from neutral to flexion posture, 8.4° and -2.5 mm (cranial displacement) for the change from neutral to extension posture, and 32.2° and 11.64 mm (caudal displacement) for the change from extension to flexion posture. The ΔLSA strongly correlated (ρ = 0.705; 95% CI, 0.434 to 0.859) with displacement of the CM. CONCLUSIONS AND CLINICAL RELEVANCE: The use of dynamic MRI, compared with conventional MRI, will better help to characterize clinically normal and abnormal features of the lumbosacral region of the vertebral column and associated spinal cord during postural changes. Further, when limited translocation of the CM is evident on dynamic MRI, veterinarians should suspect underlying lumbosacral pathophysiologic processes or anatomic abnormalities.

Point shear wave elastography of the liver in healthy adult cats.

OBJECTIVE: To evaluate stiffness of the liver parenchyma in healthy adult cats by means of point shear wave elastography (PSWE). ANIMALS: 18 client-owned adult (1- to 6-year-old) healthy cats. PROCEDURES: Echogenicity and echotexture of the liver parenchyma were assessed by means of conventional B-mode ultrasonography. The shear wave velocity (Vs) of the right and left portions of the liver were measured by means of PSWE. RESULTS: B-mode ultrasonography revealed no abnormalities in echotexture or echogenicity of the liver parenchyma in any cat. Mean (95% CI) Vs in the liver parenchyma was 1.46 m/s (1.36 to 1.55 m/s) for the right portion, 1.36 m/s (1.26 to 1.47 m/s) for the left portion, and 1.43 m/s (1.35 to 1.51 m/s) overall. The difference in mean Vs between the 2 portions of the liver was significant. No significant correlation was found between Vs and body weight or between Vs and the depth at which this variable was measured. CONCLUSIONS AND CLINICAL RELEVANCE: Quantitative PSWE of the liver was feasible in healthy adult cats. The obtained values for Vs may be useful for interpretation of and comparison with values measured in cats with liver disease. Additional research is needed to explore the potential usefulness of PSWE for diagnostic purposes.

Methylglyoxal-Induced Dysfunction in Brain Endothelial Cells via the Suppression of Akt/HIF-1α Pathway and Activation of Mitophagy Associated with Increased Reactive Oxygen Species.

Methylglyoxal (MG) is a dicarbonyl compound, the level of which is increased in the blood of diabetes patients. MG is reported to be involved in the development of cerebrovascular complications in diabetes, but the exact mechanisms need to be elucidated. Here, we investigated the possible roles of oxidative stress and mitophagy in MG-induced functional damage in brain endothelial cells (ECs). Treatment of MG significantly altered metabolic stress as observed by the oxygen-consumption rate and barrier-integrity as found in impaired trans-endothelial electrical resistance in brain ECs. The accumulation of MG adducts and the disturbance of the glyoxalase system, which are major detoxification enzymes of MG, occurred concurrently. Reactive oxygen species (ROS)-triggered oxidative damage was observed with increased mitochondrial ROS production and the suppressed Akt/hypoxia-inducible factor 1 alpha (HIF-1α) pathway. Along with the disturbance of mitochondrial bioenergetic function, parkin-1-mediated mitophagy was increased by MG. Treatment of N-acetyl cysteine significantly reversed mitochondrial damage and mitophagy. Notably, MG induced dysregulation of tight junction proteins including occludin, claudin-5, and zonula occluden-1 in brain ECs. Here, we propose that diabetic metabolite MG-associated oxidative stress may contribute to mitochondrial damage and autophagy in brain ECs, resulting in the dysregulation of tight junction proteins and the impairment of permeability.

Evaluation of a dual-purpose contrast medium for radiography and ultrasonography of the small intestine in dogs.

OBJECTIVE: To evaluate a contrast medium that could be used for radiographic and ultrasonographic assessment of the small intestine in dogs. ANIMALS: 8 healthy adult Beagles. PROCEDURES: Carboxymethylcellulose (CMC; 0.5% solution) was combined with iohexol (300 mg of iodine/mL) to yield modified contrast medium (MCM). Dogs were orally administered the first of 3 MCMs (10 mL/kg [9.5 mL of CMC/kg plus 0.5 mL of iohexol/kg]). Radiographic and ultrasonographic assessment of the small intestine followed 10 minutes after administration and every 10 minutes thereafter, until MCM was seen within the ascending colon. Minimally, 1 week elapsed between dosing of subsequent MCMs (10 mL/kg [9 mL of CMC/kg plus 1 mL of iohexol/kg and 8.5 mL of CMC/kg plus 1.5 mL of iohexol/kg]) and repeated radiography and ultrasonography. RESULTS: Radiographic contrast enhancement of the small intestine was best with MCM that combined 8.5 mL of CMC/kg and 1.5 mL of iohexol/kg. Mean small intestinal transit time for all MCMs was 86 minutes. All MCMs did not interfere with ultrasonographic assessment of the small intestine and may have improved visualization of the far-field small intestinal walls. CONCLUSIONS AND CLINICAL RELEVANCE: An MCM that combined 8.5 mL of 0.5% CMC/kg and 1.5 mL of iohexol/kg could be an alternative to barium or iohexol alone for contrast small intestinal radiography in dogs, especially when abdominal ultrasonography is to follow contrast radiography.

Feasibility and Reliability of Two-Dimensional Shear-Wave Elastography of the Liver of Clinically Healthy Cats.

Given the broad overlap of normal and abnormal liver tissue in the subjective evaluation of the liver in conventional B-mode ultrasonography, there is a need for a non-invasive and quantitative method for the diagnosis of liver disease. Novel two-dimensional shear-wave elastography (2-D SWE) can measure tissue stiffness by propagation of the shear wave induced using acoustic radiation force impulse in real time. To the best of our knowledge, two-dimensional shear-wave measurement of the liver in cats has not been reported to date. This study assessed the feasibility, reliability, normal values, and related influencing factors of 2-D SWE for assessment of the feline liver without anesthesia and breath-holding. Two-dimensional shear-wave ultrasonography was performed by two evaluators at the right and left sides of the liver. Twenty-nine client-owned clinically healthy adult cats were included. The means and standard deviations for the shear-wave speed and stiffness in the right liver were 1.52 ± 0.13 m/s and 6.94 ± 1.26 kPa, respectively, and those for the left liver were 1.61 ± 0.15 m/s and 7.90 ± 1.47 kPa, respectively. Shear-wave speed (P = 0.005) and stiffness (P = 0.002) were significantly lower in the right liver when compared to the left. The intraclass correlation value for liver stiffness was 0.835 and 0.901 for the right and left liver, respectively, indicating high interobserver agreement. Age, weight, body condition score (BCS), gabapentin administration, and measurement depths were not significantly correlated with liver stiffness or elastography measurements (P > 0.05). Our findings suggest that 2-D SWE measurements of the liver are not influenced significantly by age, weight, or BCS and can be reliably performed without anesthesia and breath-holding in cats. The values determined here can help form the basis for reference elastography values for evaluation of the feline liver.

Autophagy-mediated occludin degradation contributes to blood-brain barrier disruption during ischemia in bEnd.3 brain endothelial cells and rat ischemic stroke models.

BACKGROUND: The blood-brain barrier (BBB) maintains homeostasis of the brain environment by tightly regulating the entry of substances from systemic circulation. A breach in the BBB results in increased permeability to potentially toxic substances and is an important contributor to amplification of ischemic brain damage. The precise molecular pathways that result in impairment of BBB integrity remain to be elucidated. Autophagy is a degradation pathway that clears damaged or unnecessary proteins from cells. However, excessive autophagy can lead to cellular dysfunction and death under pathological conditions. METHODS: In this study, we investigated whether autophagy is involved in BBB disruption in ischemia, using in vitro cells and in vivo rat models. We used brain endothelial bEnd.3 cells and oxygen glucose deprivation (OGD) to simulate ischemia in culture, along with a rat ischemic stroke model to evaluate the role of autophagy in BBB disruption during cerebral ischemia. RESULTS: OGD 18 h induced cellular dysfunction, and increased permeability with degradation of occludin and activation of autophagy pathways in brain endothelial cells. Immunostaining revealed that occludin degradation is co-localized with ischemic autophagosomes. OGD-induced occludin degradation and permeability changes were significantly decreased by inhibition of autophagy using 3-methyladenine (3-MA). Enhanced autophagic activity and loss of occludin were also observed in brain capillaries isolated from rats with middle cerebral artery occlusion (MCAO). Intravenous administration of 3-MA inhibited these molecular changes in brain capillaries, and recovered the increased permeability as determined using Evans blue. CONCLUSIONS: Our findings provide evidence that autophagy plays an important role in ischemia-induced occludin degradation and loss of BBB integrity.

LRP-1 functionalized polymersomes enhance the efficacy of carnosine in experimental stroke.

Stroke is one of the commonest causes of death with limited treatment options. L-Carnosine has shown great promise as a neuroprotective agent in experimental stroke, but translation to the clinic is impeded by the large doses needed. We developed and evaluated the therapeutic potential of a novel delivery vehicle which encapsulated carnosine in lipoprotein receptor related protein-1 (LRP-1)-targeted functionalized polymersomes in experimental ischemic stroke. We found that following ischemic stroke, polymersomes encapsulating carnosine exhibited remarkable neuroprotective effects with a dose of carnosine 3 orders of magnitude lower than free carnosine. The LRP-1-targeted functionalization was essential for delivery of carnosine to the brain, as non-targeted carnosine polymersomes did not exhibit neuroprotection. Using Cy3 fluorescence in vivo imaging, we showed that unlike non-targeted carnosine polymersomes, LRP-1-targeted carriers accumulated in brain in a time dependent manner. Our findings suggest that these novel carriers have the ability to deliver neuroprotective cargo effectively to the brain.

Reduced metabolic activity of gut microbiota by antibiotics can potentiate the antithrombotic effect of aspirin.

In this study, we investigated the effects of antibiotics on the pharmacological effects of aspirin. The antithrombotic activity of aspirin was evaluated after antibiotic treatment using tail bleeding assay. The pyrosequencing analysis and selective medium culture assay were performed to investigate the alterations in gut microbiota. In addition, the in vitro metabolism assay with fecal suspension and in vivo pharmacokinetic experiments with antibiotic treatment were conducted. Ampicillin treatment significantly prolonged the bleeding time in aspirin-dosed rats. Oral administration of ampicillin significantly reduced gut microbial aspirin-metabolizing activity by 67.0% in rats. Furthermore, systemic exposure to aspirin and its primary metabolite (M1) was significantly increased in ampicillin-treated rats. The results from the pyrosequencing and selective medium culture with rat fecal samples revealed that ampicillin treatment led to the changes of the amounts and composition profile of gut microbiota. These findings suggest that co-administration of antibiotics can modulate the metabolism and pharmacokinetics of aspirin via suppression of metabolic activity of gut microbiota, which could potentiate the therapeutic potency of aspirin.