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There is a growing interest in dietary therapeutic strategies to combat oxidative stress-induced damage to the Central Nervous System (CNS), which is associated with a number of pathophysiological processes, including Alzheimer’s and Parkinson’s diseases and cerebrovascular diseases. Identifying the mechanisms associated with phenolic neuroprotection has been delayed by the lack of information concerning the ability of these compounds to enter the CNS. The aim of this study was to evaluate the transmembrane transport of flavonoids across RBE-4 cells (an immortalized cell line of rat cerebral capillary endothelial cells) and the effect of ethanol on this transport. The detection and quantification of all of the phenolic compounds in the studied samples (basolateral media) was performed using a HPLC-DAD (Diode Array Detector). All of the tested flavonoids (catechin, quercetin and cyanidin-3-glucoside) passed across the RBE-4 cells in a time-dependent manner. This transport was not influenced by the presence of 0.1% ethanol. In conclusion, the tested flavonoids were capable of crossing this blood-brain barrier model.
Arginine vasopressin (AVP) has been shown to promote the disruption of the blood-brain barrier (BBB) and the formation of edema in various animal models of brain injury. However, the source(s) of this AVP have not been identified. Since the cerebral cortex was considerably affected in some of these brain injury models, we sought to determine if AVP was produced in the cerebral cortex, and, if so, whether or not this cortical AVP expression was up regulated after injury. In the present study, a controlled cortical impact model of traumatic brain injury (TBI) in rats was used, and the temporal changes in expression of AVP and its V1a receptor were analyzed by real-time reverse-transcriptase polymerase chain reaction. The expression of AVP and its V1a receptor in the ipsilateral cortex adjacent to the lesion area was significantly up regulated between 4 h and 1day post-TBI. The maximum increase in mRNA for AVP (4.3-fold) and its receptor (2.6-fold) in the ipsilateral vs. contralateral cortex was observed at 6 h post-TBI. Compared to sham-injured rats, no statistically significant changes in expression of AVP or its receptor were found in the contralateral cortex. These results suggest that the cerebral cortex is an important source of AVP in the injured brain, and the parallel increase in the expression of AVP and its cognate receptor may act to augment the actions of AVP related to promoting the disruption of the BBB and the formation of post-traumatic edema.
Changes in the large neutral amino acid (LNAA) transport across the blood-brain barrier (BBB) is thought to contribute to brain dysfunction in a number of clinical conditions, including phenylketonuria, acute liver failure, and sepsis. Here, we present a novel approach for estimating BBB permeability and the LNAA concentrations in brain extracellular fluid, by demonstrating that they can be mathematically derived on the basis of kinetic constants of the BBB available from the literature, if cerebral blood flow and the arterial and jugular venous LNAA concentrations are known. While it is well known that the permeability surface area product of the BBB to a LNAA from blood to brain (PS1) can be calculated from the arterial LNAA concentrations and kinetic constants of the BBB, we demonstrate that the permeability surface area product from brain to blood (PS2) can be calculated by deriving the substrate activity of the saturable transporter from the kinetic constants and arterial and jugular venous LNAA concentrations, and that the concentration of the LNAA in brain extracellular fluid can then be determined. This approach is methodically simple, and may be useful for assessing the transcerebral exchange kinetics of LNAAs in future human-experimental and clinical studies.
Many anti-cancer and antiviral drugs currently used are either unable, or inefficient in their ability to pass through the blood brain-barrier and to enter and maintain therapeutic drug levels in brain. The low bioavailability of these drugs is a limiting factor in their use. In order to overcome these limitations, we ester-linked various anti-cancer and antiviral drugs to ceramide and phosphatidylcholine and created prodrugs possessing therapeutic attributes lacking in the parent compounds. This resulted in greater cellular uptake and prolonged retention of these prodrugs in vitro. Likewise, prodrug concentration was greater and retention time longer than the parent drug in the brain, testes and thymus of mice. Another major goal in drug development is discovering compounds that have efficacy against a specific microorganism or virus without significant side effects. For example, many potentially good drugs cannot be used because they are either toxic to uninfected cells or they cannot be restricted to a certain part of the body. If a drug could remain inert unless and until it is inside an infected cell, many of the common problems associated with drug treatments would be solved. In an attempt to address this problem we are developing a method by which a drug will be released only in cells that are infected with a particular microorganism or virus. The methodology makes use of the fact that microparticles are ingested by macrophages. Cell-specific treatment can be achieved by combining a drug with a microparticle using microorganism-specific enzyme substrates. Thus, release of active drug will occur only in the presence of enzymes specific to the target virus or microorganism. In the uninfected macrophage drug remains bound to the microparticle and is inactive. In the infected cell active drug is released by enzymatic hydrolysis. Potential applications for this technology include all diseases in which pathogens are resident in macrophages and other phagocytic cells.
The present study was conducted to immunolocalize 3b-hydroxysteroid dehydrogenase (3β-HSD), an enzyme metabolizing pregnenolone to progesterone in the choroid plexus of the lateral ventricle in sheep, as well as to measure progesterone concentration in cerebrospinal fluid (CSF) and plasma using radioimmunoassay (RIA). Akkaraman breed rams (n = 16) and ewes (n = 16) were utilized in the study. 3β-HSD was immunolocalized in choroid epithelial cells of the choroid plexus with an apparent cytoplasmic immunoreactivity. Progesterone was detected in CSF with no significant differences between the ewes (0.76 ± 0.14 ng/mL) and rams (0.74 ± 0.13 ng/mL) (p > 0.05). However, the plasma progesterone concentration in the ewes (0.27 ± 0.04 ng/mL) was significantly higher than that of the rams (0.11 ± 0.02 ng/mL) (p < 0.001). Consequently, CSF in sheep contains progesterone in significant levels. As evidenced by 3β-HSD immunoreactivity, choroid epithelial cells may be a site of progesterone synthesis in sheep.
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