Recent Research Articles from UNTHSC

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Lymphatic pump treatment repeatedly enhances the lymphatic and immune systems.

Fri, 08/01/2014 - 4:05am
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Lymphatic pump treatment repeatedly enhances the lymphatic and immune systems.

Lymphat Res Biol. 2013 Dec;11(4):219-26

Authors: Schander A, Padro D, King HH, Downey HF, Hodge LM

Abstract
BACKGROUND: Osteopathic practitioners utilize manual therapies called lymphatic pump techniques (LPT) to treat edema and infectious diseases. While previous studies examined the effect of a single LPT treatment on the lymphatic system, the effect of repeated applications of LPT on lymphatic output and immunity has not been investigated. Therefore, the purpose of this study was to measure the effects of repeated LPT on lymphatic flow, lymph leukocyte numbers, and inflammatory mediator concentrations in thoracic duct lymph (TDL).
METHODS AND RESULTS: The thoracic ducts of five mongrel dogs were cannulated, and lymph samples were collected during pre-LPT, 4 min of LPT, and 2 hours post-LPT. A second LPT (LPT-2) was applied after a 2 hour rest period. TDL flow was measured, and TDL were analyzed for the concentration of leukocytes and inflammatory mediators. Both LPT treatments significantly increased TDL flow, leukocyte count, total leukocyte flux, and the flux of interleukin-8 (IL-8), keratinocyte-derived chemoattractant (KC), nitrite (NO2(-)), and superoxide dismutase (SOD). The concentration of IL-6 increased in lymph over time in all experimental groups; therefore, it was not LPT dependent.
CONCLUSION: Clinically, it can be inferred that LPT at a rate of 1 pump per sec for a total of 4 min can be applied every 2 h, thus providing scientific rationale for the use of LPT to repeatedly enhance the lymphatic and immune system.

PMID: 24364845 [PubMed - indexed for MEDLINE]

Monitoring retinal morphologic and functional changes in mice following optic nerve crush.

Fri, 08/01/2014 - 4:05am
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Monitoring retinal morphologic and functional changes in mice following optic nerve crush.

Invest Ophthalmol Vis Sci. 2014;55(6):3766-74

Authors: Liu Y, McDowell CM, Zhang Z, Tebow HE, Wordinger RJ, Clark AF

Abstract
PURPOSE: We characterized the morphologic and functional changes in optic nerve crushed mice and evaluated electroretinogram (ERG) responses as tools to monitor retinal ganglion cell (RGC) dysfunction.
METHODS: We performed optic nerve crush (ONC) unilaterally in adult BALB/cJ mice. The neuronal loss in the RGC layer (GCL) and superior colliculus (SC) was determined by Nissl staining. Retinal thickness was assessed by spectral-domain optical coherence tomography (SD-OCT) imaging. Retinal function was determined by pattern ERG and full-field flash ERG. Responses of pattern ERG, positive scotopic threshold response (pSTR), scotopic oscillatory potentials (OPs), and photopic negative response (PhNR) were analyzed.
RESULTS: The ONC induced progressive neuronal loss in GCL and contralateral SC starting from 7 and 28 days following ONC, respectively. A linear correlation was observed between combined thickness of nerve fiber layer (NFL), GCL, and inner plexiform layer (IPL) imaged by SD-OCT and cell numbers in GCL. Only half of the normal BALB/cJ mice exhibited pattern ERG response, which was smaller and later compared to C57BL/6J mice. The ONC reduced pattern ERG and pSTR, but the reduction of pattern ERG was earlier than pSTR, preceding the anatomical cell loss in the GCL. The PhNR and scotopic OPs were not affected by ONC.
CONCLUSIONS: The SD-OCT and ERG can be used to monitor noninvasively retinal morphologic and functional changes induced by ONC. Pattern ERG and pSTR are able to detect early RGC dysfunction, but pattern ERG exhibits higher sensitivity. Our results support the use of these tools in studies using the mouse ONC model.

PMID: 24854856 [PubMed - indexed for MEDLINE]

Involvement of AP-1 and C/EBPβ in upregulation of endothelin B (ETB) receptor expression in a rodent model of glaucoma.

Wed, 07/30/2014 - 4:05am
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Involvement of AP-1 and C/EBPβ in upregulation of endothelin B (ETB) receptor expression in a rodent model of glaucoma.

PLoS One. 2013;8(11):e79183

Authors: He S, Minton AZ, Ma HY, Stankowska DL, Sun X, Krishnamoorthy RR

Abstract
Previous studies showed that the endothelin B receptor (ETB) expression was upregulated and played a key role in neurodegeneration in rodent models of glaucoma. However, the mechanisms underlying upregulation of ETB receptor expression remain largely unknown. Using promoter-reporter assays, the 1258 bp upstream the human ETB promoter region was found to be essential for constitutive expression of ETB receptor gene in human non-pigmented ciliary epithelial cells (HNPE). The -300 to -1 bp and -1258 to -600 bp upstream promoter regions of the ETB receptor appeared to be the key binding regions for transcription factors. In addition, the crucial AP-1 binding site located at -615 to -624 bp upstream promoter was confirmed by luciferase assays and CHIP assays which were performed following overexpression of c-Jun in HNPE cells. Overexpression of either c-Jun or C/EBPβ enhanced the ETB receptor promoter activity, which was reflected in increased mRNA and protein levels of ETB receptor. Furthermore, knock-down of either c-Jun or C/EBPβ in HNPE cells was significantly correlated to decreased mRNA levels of both ETB and ETA receptor. These observations suggest that c-Jun and C/EBPβ are important for regulated expression of the ETB receptor in HNPE cells. In separate experiments, intraocular pressure (IOP) was elevated in one eye of Brown Norway rats while the corresponding contralateral eye served as control. Two weeks of IOP elevation produced increased expression of c-Jun and C/EBPβ in the retinal ganglion cell (RGC) layer from IOP-elevated eyes. The mRNA levels of c-Jun, ETA and ETB receptor were upregulated by 2.2-, 3.1- and 4.4-fold in RGC layers obtained by laser capture microdissection from retinas of eyes with elevated IOP, compared to those from contralateral eyes. Taken together, these data suggest that transcription factor AP-1 plays a key role in elevation of ETB receptor in a rodent model of ocular hypertension.

PMID: 24265756 [PubMed - indexed for MEDLINE]

Bilorrhea Secondary to Bronchobiliary Fistula.

Sat, 07/26/2014 - 4:06am

Bilorrhea Secondary to Bronchobiliary Fistula.

Int Surg. 2014 July-August;99(4):438-441

Authors: Olivencia-Yurvati AH, Rollins C

Abstract
Abstract Bronchobiliary fistula (BBF) is a rare condition which occurs most commonly as a complication of hydatid cyst liver disease. The following report describes a patient who presented with biliptysis 6 months following decortication of an empyema that had occurred following partial hepatectomy of a colon cancer metastasis. This is the only case to our knowledge that describes the presentation of a BBF in this context. The patient was diagnosed with BBF and successfully underwent open thoracotomy for fistulectomy and repair.

PMID: 25058780 [PubMed - as supplied by publisher]

Two-week Normobaric Intermittent-Hypoxia Exposures Enhance Oxyhemoglobin Equilibrium and Cardiac Responses during Hypoxemia.

Sat, 07/26/2014 - 4:06am

Two-week Normobaric Intermittent-Hypoxia Exposures Enhance Oxyhemoglobin Equilibrium and Cardiac Responses during Hypoxemia.

Am J Physiol Regul Integr Comp Physiol. 2014 Jul 23;

Authors: Zhang P, Downey HF, Chen S, Shi X

Abstract
Intermittent-hypoxia (IH) is extensively applied to challenge cardiovascular and respiratory function, and to induce physiological acclimatization. The purpose of this study was to test the hypothesis that oxyhemoglobin equilibrium and tachycardiac responses during hypoxemia were enhanced after 14-day IH exposures. Normobaric-poikilocapnic hypoxia was induced with inhalation of 10% O2 for 5-6 min interspersed with 4-min recovery on eight non-smokers. Heart rate (HR), arterial O2 saturation (SaO2) and end-tidal O2 (PETO2) were continuously monitored during cyclic normoxia and hypoxia. These variables were compared during the 1st and 5th hypoxic bouts between Day 1 and Day 14. There was a rightward shift in the oxyhemoglobin equilibrium response following 14-day IH exposures, as indicated by the greater PETO2 (an index of arterial PO2) at 50% of SaO2 (P50) on Day 14 compared to Day 1: 33.9±1.5 vs 28.2±1.3 mmHg (P =0.005) during the 1st hypoxic bout and 39.4±2.4 vs 31.4±1.5 mmHg (P =0.006) during the 5th hypoxic bout; and by the augmented gains of ∆SaO2/∆PETO2 (i.e., de-oxygenation) during PETO2 from 65 to 40 mmHg in the 1st (1.12±0.08 vs 0.80±0.02 %/mmHg, P =0.001) and the 5th (1.76±0.31 vs 1.05±0.06 %/mmHg, P =0.024) hypoxic bouts. Repetitive IH exposures attenuated (P =0.049) the tachycardiac response to hypoxia while significantly enhanced normoxic R-R interval variability in low-frequency and high-frequency spectra without changes in arterial blood pressure at rest or during hypoxia. We conclude that 14-day IH exposures enhance arterial O2 delivery and improve vagal control of HR during hypoxic hypoxemia.

PMID: 25056104 [PubMed - as supplied by publisher]

Modulation of the rate of retinal degeneration in T17M RHO mice by reprogramming the unfolded protein response.

Sat, 07/26/2014 - 4:06am
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Modulation of the rate of retinal degeneration in T17M RHO mice by reprogramming the unfolded protein response.

Adv Exp Med Biol. 2014;801:455-62

Authors: Choudhury S, Nashine S, Bhootada Y, Kunte MM, Gorbatyuk O, Lewin AS, Gorbatyuk M

Abstract
The goal of this study is to validate whether reprogramming of the UPR via modulation of pro-apoptotic caspase-7 and CHOP proteins could be an effective approach to slow down the rate of retinal degeneration in ADRP mice. In order to pursue our goal we created the T17M RHO CASP7 and T17M RHO CHOP mice to study the impact of the CASP7 or CHOP ablations in T17M RHO retina by ERG, SD-OCT, histology and western blot analysis. The scotopic ERG demonstrated that the ablation of the CASP7 in T17M RHO retina leads to significant preservation of the function of photoreceptors compared to control. Surprisingly, the ablation of pro-apoptotic CHOP protein in T17M RHO mice led to a more severe form of retinal degeneration. Results of the SD-OCT and histology were in agreement with the ERG data. The further analysis demonstrated that the preservation of the structure and function or the acceleration of the onset of the T17M RHO photoreceptor degeneration occurred via reprogramming of the UPR. In addition, the CASP7 ablation leads to the inhibition of cJUN mediated apoptosis, while the ablation of CHOP induces an increase in the HDAC. Thus, manipulation with the UPR requires careful examination in order to achieve a therapeutic effect.

PMID: 24664731 [PubMed - indexed for MEDLINE]

Reduction of stutter ratios in short tandem repeat loci typing of low copy number DNA samples.

Fri, 07/25/2014 - 4:05am
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Reduction of stutter ratios in short tandem repeat loci typing of low copy number DNA samples.

Forensic Sci Int Genet. 2014 Jan;8(1):213-8

Authors: Seo SB, Ge J, King JL, Budowle B

Abstract
Increased height of stutter peaks is a phenomenon with low copy number (LCN) short tandem repeat (STR) typing that can impact interpretation. An alternative strategy of lowering the annealing/extension temperature (LT) at 56 °C was designed to attempt to decrease the heights of stutter peaks. STR typing results were generated in terms of stutter ratios using LT-PCR conditions and compared with data obtained using standard (STD) PCR conditions. DNA samples ranging from 100 to 25 pg were amplified using reagents contained in the AmpFℓSTR Identifiler PCR Amplification or AmpFℓSTR Identifiler Plus PCR Amplification kits with 32 or 34 PCR cycles. Stutter ratios decreased by an average of 14.7%, 14.9% and 18.1% at 100, 50 and 25 pg of template DNA under LT conditions compared with those of STD conditions in the Identifiler Kit amplified samples. The LT conditions also decreased average stutter ratios by 13.3% compared with those of STD conditions in the Identifiler Plus Kit amplified samples. The overall PCR efficiency obtained with STD and LT conditions with the two STR kits was comparable in terms of the number of detected alleles, peak heights and peak height ratios. These results support the hypothesis that a lower temperature annealing/extension step reduces the likelihood of slippage during PCR by enhancing the stability of the DNA polymerase/template DNA complex or the stability of the generated duplex than the conditions of the standard extension step. This stability in turn would result in lower stutter ratios.

PMID: 24315611 [PubMed - indexed for MEDLINE]

Low serum zinc is associated with elevated risk of cadmium nephrotoxicity.

Thu, 07/24/2014 - 4:04am

Low serum zinc is associated with elevated risk of cadmium nephrotoxicity.

Environ Res. 2014 Jul 17;134C:33-38

Authors: Lin YS, Ho WC, Caffrey JL, Sonawane B

Abstract
BACKGROUND: Despite animal evidence suggests that zinc modulates cadmium nephrotoxicity, limited human data are available.
OBJECTIVE: To test the hypothesis that low serum zinc concentrations may increase the risk of cadmium-mediated renal dysfunction in humans.
METHODS: Data from 1545 subjects aged 20 or older in the National Health and Nutrition Examination Survey (NHANES), 2011-2012 were analyzed. Renal function was defined as impaired when estimated glomerular filtration rate (eGFR) fell below 60ml/min/1.73m(2) and/or the urinary albumin-to-creatinine ratio surpassed 2.5 in men and 3.5mg/mmol in women.
RESULTS: Within the study cohort, 117 subjects had reduced eGFR and 214 had elevated urinary albumin. After adjusting for potential confounders, subjects with elevated blood cadmium (>0.53μg/L) were more likely to have a reduced eGFR (odds ratio [OR]=2.21, 95% confidence interval [CI]: 1.09-4.50) and a higher urinary albumin (OR=2.04, 95% CI: 1.13-3.69) than their low cadmium (<0.18μg/L) peers. In addition, for any given cadmium exposure, low serum zinc is associated with elevated risk of reduced eGFR (OR=3.38, 95% CI: 1.39-8.28). A similar increase in the odds ratio was observed between declining serum zinc and albuminuria but failed to reach statistical significance. Those with lower serum zinc/blood cadmium ratios were likewise at a greater risk of renal dysfunction (p<0.01).
CONCLUSIONS: This study results suggest that low serum zinc concentrations are associated with an increased risk of cadmium nephrotoxicity. Elevated cadmium exposure is global public health issue and the assessment of zinc nutritional status may be an important covariate in determining its effective renal toxicity.

PMID: 25042034 [PubMed - as supplied by publisher]

Characterization of [(3) H]LS-3-134, a Novel Arylamide Phenylpiperazine D3 Dopamine Receptor Selective Radioligand.

Thu, 07/24/2014 - 4:04am

Characterization of [(3) H]LS-3-134, a Novel Arylamide Phenylpiperazine D3 Dopamine Receptor Selective Radioligand.

J Neurochem. 2014 Jul 18;

Authors: Rangel-Barajas C, Malik M, Taylor M, Neve KA, Mach RH, Luedtke RR

Abstract
LS-3-134 is a substituted N-phenylpiperazine derivative that has been reported to exhibit a) high-affinity binding (Ki value 0.2 nM) at human D3 dopamine receptors, b) >100-fold D3 vs. D2 dopamine receptor subtype binding selectivity and c) low-affinity binding (Ki values >5,000 nM) at sigma 1 and sigma 2 receptors. Based upon a forskolin-dependent activation of the adenylyl cyclase inhibition assay, LS-3-134 is a weak partial agonist at both D2 and D3 dopamine receptor subtypes (29% and 35% of full agonist activity, respectively). In this study, [(3) H]-labeled LS-3-134 was prepared and evaluated to further characterize its use as a D3 dopamine receptor selective radioligand. Kinetic and equilibrium radioligand binding studies were performed. This radioligand rapidly reaches equilibrium (10-15 min at 37°C) and binds with high affinity to both human (Kd = 0.06 ± 0.01 nM) and rat (Kd = 0.2 ± 0.02 nM) D3 receptors expressed in HEK-293 cells. Direct and competitive radioligand binding studies using rat caudate and nucleus accumbens tissue indicate that [(3) H]LS-3-134 selectively binds a homogeneous population of binding sites with a dopamine D3 receptor pharmacological profile. Based upon these studies we propose that [(3) H]LS-3-134 represents a novel D3 dopamine receptor selective radioligand that can be used for studying the expression and regulation of the D3 dopamine receptor subtype. This article is protected by copyright. All rights reserved.

PMID: 25041389 [PubMed - as supplied by publisher]

Lenticular mitoprotection. Part B: GSK-3β and regulation of mitochondrial permeability transition for lens epithelial cells in atmospheric oxygen.

Thu, 07/24/2014 - 4:04am
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Lenticular mitoprotection. Part B: GSK-3β and regulation of mitochondrial permeability transition for lens epithelial cells in atmospheric oxygen.

Mol Vis. 2013;19:2451-67

Authors: Brooks MM, Neelam S, Cammarata PR

Abstract
PURPOSE: Loss of integrity of either the inner or outer mitochondrial membrane results in the dissipation of the mitochondrial electrochemical gradient that leads to mitochondrial membrane permeability transition (mMPT). This study emphasizes the role of glycogen synthase kinase 3beta (GSK-3β) in maintaining mitochondrial membrane potential, thus preventing mitochondrial depolarization (hereafter termed mitoprotection). Using 3-(2,4-dichlorophenyl)-4-(1-methyl-1H-indol-3-yl)-1H-pyrrole-2,5-dione (SB216763), an inhibitor of GSK-3β, and drawing a distinction between it and 1,4-diamino-2,3-dicyano-1,4-bis[2-aminophenylthio] butadiene (UO126), an inhibitor of extracellular-signal-regulated kinase (ERK) phosphorylation, the means by which GSK-3β influences mitoprotection in cultured human lens epithelial (HLE-B3) cells and normal, secondary cultures of bovine lens epithelial cells, maintained in atmospheric oxygen, was investigated.
METHODS: Virally transfected human lens epithelial cells (HLE-B3) and normal cultures of bovine lens epithelial cells were exposed to acute hypoxic conditions (about 1% O2) followed by exposure to atmospheric oxygen (about 21% O2). Specific antisera and western blot analysis was used to examine the state of phosphorylation of ERK and GSK-3β, as well as the phosphorylation of a downstream substrate of GSK-3β, glycogen synthase (GS, useful in monitoring GSK-3β activity). The potentiometric dye, 1H-benzimidazolium-5,6-dichloro-2-[3-(5,6-dichloro-1,3-diethyl-1,3-dihydro-2H-benzimidazol-2-ylidene)-1-propenyl]-1,3-diethyl-iodide (JC-1), was used to monitor mitochondrial depolarization upon exposure of inhibitor treatment relative to the control cells (mock inhibition) in atmospheric oxygen. Caspase-3 activation was scrutinized to determine whether mitochondrial depolarization inevitably leads to apoptosis.
RESULTS: Treatment of HLE-B3 cells with SB216763 (12 µM) inactivated GSK-3β activity as verified by the enzyme's inability to phosphorylate its substrate, GS. SB216763-treated cells were not depolarized relative to the control cells as demonstrated with JC-1 fluorescent dye analysis. The HLE-B3 cells treated with UO126, which similarly blocked phosphorylation of GS, were nevertheless prone to mMPT relative to the control cells. Western blot analysis determined that Bcl-2-associated X (BAX) levels were unchanged for SB216763-treated or UO126-treated HLE-B3 cells when compared to their respective control cells. However, unlike the SB216763-treated cells, the UO126-treated cells showed a marked absence of Bcl-2, as well as phosphorylated Bcl-2 relative to the controls. UO126 treatment of bovine lens epithelial cells showed similar results with pBcl-2 levels, while the Bcl-2 content appeared unchanged relative to the control cells. HLE-B3 and normal bovine lens cell cultures showed susceptibility to mMPT associated with the loss of pBcl-2 by UO126 treatment.
CONCLUSIONS: MITOCHONDRIAL DEPOLARIZATION MAY OCCUR BY ONE OF TWO KEY OCCURRENCES: interruption of the electrochemical gradient across the inner mitochondrial membrane resulting in mMPT or by disruption of the integrity of the inner or outer mitochondrial membrane. The latter scenario is generally tightly regulated by members of the Bcl-2 family of proteins. Inhibition of GSK-3β activity by SB216763 blocks mMPT by preventing the opening of the mitochondrial permeability transition pore. UO126, likewise, inhibits GSK-3β activity, but unlike SB216763, inhibition of ERK phosphorylation induces the loss of intracellular pBcl-2 levels under conditions where intracellular BAX levels remain constant. These results suggest that the lenticular mitoprotection normally afforded by the inactivation of GSK-3β activity may, however, be bypassed by a loss of pBcl-2, an anti-apoptotic member of the Bcl-2 family. Bcl-2 prevents the translocation of BAX to the mitochondrial outer membrane inhibiting depolarization by disrupting the normal electrochemical gradient leading to mMPT.

PMID: 24319338 [PubMed - indexed for MEDLINE]

Single nucleotide polymorphism typing with massively parallel sequencing for human identification.

Thu, 07/24/2014 - 4:04am
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Single nucleotide polymorphism typing with massively parallel sequencing for human identification.

Int J Legal Med. 2013 Nov;127(6):1079-86

Authors: Seo SB, King JL, Warshauer DH, Davis CP, Ge J, Budowle B

Abstract
The Ion AmpliSeq™ HID single nucleotide polymorphism (SNP) panel, a primer pool of 103 autosomal SNPs and 33 Y-SNPs, was evaluated using the Ion 314™ Chip on the Ion PGM™ Sequencer with four DNA samples. The study focused on the sequencing of DNA at three different initial target quantities, related interpretation issues, and concordance of results with another sequencing platform, i.e., Genome Analyzer IIx. With 10 ng of template DNA, all genotypes at the 136 SNPs were detected. With 1 ng of DNA, all SNPs were detected and one SNP locus in one sample showed extreme heterozygote imbalance on allele coverage. With 100 pg of DNA, an average of 1.6 SNP loci were not detected, and an average of 4.3 SNPs showed heterozygote imbalance. The average sequence coverage was 945-600× at autosomal SNPs and 465-209× at Y-SNPs for 10 ng-100 pg of DNA. The average heterozygote allele coverage ratio was 89.6-61.8 % for 10 ng-100 pg of DNA. At 10 ng of DNA, all genotypes of the 95 SNPs shared between the two different sequencing platforms were concordant except for one SNP, rs1029047. The error was due to the misalignment of a flanking homopolymer. Overall, the data support that genotyping a large battery of SNPs is feasible with massively parallel sequencing. With barcode systems, better allele balance, and specifically designed alignment software, a more comprehensive rapid genotyping and more cost-effective results may be obtained from multiple samples in one analysis than are possible with current typing and capillary electrophoresis systems.

PMID: 23736940 [PubMed - indexed for MEDLINE]

5-hydroxytryptamine-mediated neurotransmission modulates spontaneous and vagal-evoked glutamate release in the nucleus of the solitary tract effect of uptake blockade.

Tue, 07/22/2014 - 4:04am
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5-hydroxytryptamine-mediated neurotransmission modulates spontaneous and vagal-evoked glutamate release in the nucleus of the solitary tract effect of uptake blockade.

J Pharmacol Exp Ther. 2014 May;349(2):288-96

Authors: Hosford PS, Mifflin SW, Ramage AG

Abstract
The effect of blockade of either 5-hydroxytryptamine (5-HT)/serotonin transporter (SERT) with citalopram or the organic cation transporter 3 (OCT3)/plasma membrane monoamine transporter (PMAT) with decynium-22 (D-22) on spontaneous and evoked release of 5-HT in the nucleus tractus solitarius (NTS) was investigated in rat brainstem slices treated with gabazine. 5-HT release was measured indirectly by changes in the frequency and amplitude of glutamatergic miniature excitatory postsynaptic currents (mEPSCs) [in the presence of tetrodotoxin (TTX)] and evoked EPSCs. Blockade of 5-HT3 receptors with granisetron reduced, whereas the 5-HT3 agonist phenylbiguanide increased, the frequency of mEPSCs. 5-HT decreased mEPSC frequency at low concentrations and increased frequency at high concentrations. This inhibition was blocked by the 5-HT1A antagonist N-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-N-2-pyridinylcyclohexanecarboxamide (WAY-100635), which was ineffective on its own, whereas the excitation was reversed by granisetron. The addition of citalopram or D-22 caused inhibition, which was prevented by 5-HT1A blockade. Thus, in the NTS, the spontaneous release of 5-HT is able to activate 5-HT3 receptors, but not 5-HT1A receptors, as the release in their vicinity is removed by uptake. The ineffectiveness of corticosterone suggests that the low-affinity, high-capacity transporter is PMAT, not OCT3. For evoked 5-HT release, only D-22 caused an increase in the amplitude of EPSCs, with a decrease in the paired pulse ratio, and increased the number of spontaneous EPSCs after 20-Hz stimulation. Thus, for the evoked release of 5-HT, the low-affinity, high-capacity transporter PMAT, but not 5-HT transporter (5-HTT)/SERT, is important in the regulation of changes in 5-HT extracellular concentration.

PMID: 24618127 [PubMed - indexed for MEDLINE]

Establishment of human retinal microvascular endothelial cells with extended life-span.

Sat, 07/19/2014 - 4:06am
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Establishment of human retinal microvascular endothelial cells with extended life-span.

In Vivo. 2013 Nov-Dec;27(6):685-94

Authors: Kashyap MV, Ranjan AP, Shankardas J, Vishwanatha JK

Abstract
AIM: To generate and characterize a telomerase-immortalized human retinal microvascular endothelial cell (HREC) line. This cell line may be utilized as an in vitro model to study the molecular basis of several diseases of the human retina.
MATERIALS AND METHODS: Primary retinal neuronal cells were isolated and transfected with plasmid encoding full-length human telomerase reverse transcriptase (hTERT). Transfected cells were selected and characterized to determine telomerase activity, karyotype, proliferative capacity and functionality.
RESULTS: HREC-hTERT cells appear morphologically similar to primary endothelial cells and have an extended in vitro life-span. HREC-hTERT cells express the progenitor/stem cell marker nestin. They have active telomerase and a high proliferative capacity. These cells also maintain a diploid karyotype. The HREC-hTERT cells showed high colony-formation capacity and plating efficiency compared to the primary cells. These cells are capable of differentiation into neuronal and glial cell phenotypes and the differentiated cells express the astrocyte marker glial fibrillary acidic protein (GFAP) and the neuronal marker microtubule-associated protein-2 (MAP2), respectively.
CONCLUSION: The in vitro life-span of human retinal neuronal endothelial cells can be extended by ectopic expression of hTERT without altering the genetic stability and functionality of these cells. These cells will be a valuable tool to further our understanding on the role of HRECs in the human blood-retinal-barrier and in angiogenesis and neovascularization.

PMID: 24292569 [PubMed - indexed for MEDLINE]

Treatment of Acute Pulmonary Embolism: Update on Newer Pharmacologic and Interventional Strategies.

Thu, 07/17/2014 - 4:04am
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Treatment of Acute Pulmonary Embolism: Update on Newer Pharmacologic and Interventional Strategies.

Biomed Res Int. 2014;2014:410341

Authors: Pelliccia F, Schiariti M, Terzano C, Keylani AM, D'Agostino DC, Speziale G, Greco C, Gaudio C

Abstract
Acute pulmonary embolism (PE) is a common complication in hospitalized patients, spanning multiple patient populations and crossing various therapeutic disciplines. Current treatment paradigm in patients with massive PE mandates prompt risk stratification with aggressive therapeutic strategies. With the advent of endovascular technologies, various catheter-based thrombectomy and thrombolytic devices are available to treat patients with massive or submassive PE. In this paper, a variety of newer treatment strategies for PE are analyzed, with special emphasis on various interventional treatment strategies. Clinical evidence for utilizing endovascular treatment modalities, based on our institutional experience as well as a literature review, is provided.

PMID: 25025049 [PubMed - as supplied by publisher]

Validation of a Serum Screen for Alzheimer's Disease Across Assay Platforms, Species, and Tissues.

Thu, 07/17/2014 - 4:04am
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Validation of a Serum Screen for Alzheimer's Disease Across Assay Platforms, Species, and Tissues.

J Alzheimers Dis. 2014 Jul 11;

Authors: O'Bryant SE, Xiao G, Zhang F, Edwards M, German DC, Yin X, Como T, Reisch J, Huebinger RM, Graff-Radford N, Dickson D, Barber R, Hall J, O'Suilleabhain P, Grammas P

Abstract
Background: There is a significant need for rapid and cost-effective biomarkers of Alzheimer's disease (AD) for advancement of clinical practice and therapeutic trials. Objective: The aim of the current study was to cross-validate our previously published serum-based algorithm on an independent assay platform as well as validate across tissues and species. Preliminary analyses were conducted to examine the utility in distinguishing AD from non-AD neurological disease (Parkinson's disease, PD). Methods: Serum proteins from our previously published algorithm were quantified from 150 AD cases and 150 controls on the Meso Scale Discovery (MSD) platform. Serum samples were analyzed from 49 PD cases and compared to a random sample of 51 AD cases and 62 controls. Support vector machines (SVM) were used to discriminate PD versus AD versus controls. Human and AD mouse model microvessel images were quantified with HAMAMATSU imaging software. Mouse serum biomarkers were assayed via MSD. Results: Analysis of 21 serum proteins from 150 AD cases and 150 controls yielded an algorithm with sensitivity and specificity of 0.90 for correctly classifying AD. This multi-marker approach was then validated across species and tissue. Assay of the top proteins in human and AD mouse model brain microvessels correctly classified 90-100% of the samples. SVM analyses were highly accurate at distinguishing PD versus AD versus controls. Conclusions: This serum-based biomarker panel should be tested in a community-based setting to determine its utility as a first-line screen for AD and non-AD neurological diseases for primary care providers.

PMID: 25024345 [PubMed - as supplied by publisher]

In vivo Assessment of SMT19969 in the Hamster Model of Clostridium difficile Infection.

Thu, 07/17/2014 - 4:04am
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In vivo Assessment of SMT19969 in the Hamster Model of Clostridium difficile Infection.

Antimicrob Agents Chemother. 2014 Jul 14;

Authors: Weiss W, Pulse M, Vickers R

Abstract
SMT19969 (2, 2' bis(4-pyridyl)3H, 3' H 5,5 bibenzimidazole) is a novel, narrow spectrum, non-absorbable antibiotic currently in development for the treatment of Clostridium difficile infection. The comparative activity of SMT19969 and vancomycin against non-epidemic and epidemic strains of C. difficile was studied in an established hamster model. Against non-epidemic (VA11) strains, survival ranged from 80% to 95% in SMT19969 treated animals. Vancomycin exhibited 100% protection during treatment with relapse observed starting on Day 9 and 50% survival at Day 20. At 50 mg/kg, SMT19969 administered orally, qd x 5 days exhibited full protection of treated animals on dosing days and out through day 12 against epidemic strains. Vancomycin also protected during the dosing interval, but apparent relapse occurred earlier starting on day 11. SMT19969 exhibited excellent in vitro activity with an MIC of 0.25 ug/mL for all isolates. MICs for vancomycin were 2-4 fold higher at ≤0.5-1 ug/mL. All plasma samples of SMT19969 were below the limit of quantitation (25 ng/mL) at all time points, consistent with the reported lack of bioavailability of the compound. Cecal concentrations were significantly above MIC (ranging from 96μg/mL to 172μg/mL).

PMID: 25022586 [PubMed - as supplied by publisher]

Pathogenesis of Chronic Hyperglycemia: From Reductive Stress to Oxidative Stress.

Thu, 07/17/2014 - 4:04am
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Pathogenesis of Chronic Hyperglycemia: From Reductive Stress to Oxidative Stress.

J Diabetes Res. 2014;2014:137919

Authors: Yan LJ

Abstract
Chronic overnutrition creates chronic hyperglycemia that can gradually induce insulin resistance and insulin secretion impairment. These disorders, if not intervened, will eventually be followed by appearance of frank diabetes. The mechanisms of this chronic pathogenic process are complex but have been suggested to involve production of reactive oxygen species (ROS) and oxidative stress. In this review, I highlight evidence that reductive stress imposed by overflux of NADH through the mitochondrial electron transport chain is the source of oxidative stress, which is based on establishments that more NADH recycling by mitochondrial complex I leads to more electron leakage and thus more ROS production. The elevated levels of both NADH and ROS can inhibit and inactivate glyceraldehyde 3-phosphate dehydrogenase (GAPDH), respectively, resulting in blockage of the glycolytic pathway and accumulation of glycerol 3-phospate and its prior metabolites along the pathway. This accumulation then initiates all those alternative glucose metabolic pathways such as the polyol pathway and the advanced glycation pathways that otherwise are minor and insignificant under euglycemic conditions. Importantly, all these alternative pathways lead to ROS production, thus aggravating cellular oxidative stress. Therefore, reductive stress followed by oxidative stress comprises a major mechanism of hyperglycemia-induced metabolic syndrome.

PMID: 25019091 [PubMed - as supplied by publisher]

Manual therapy, exercise, and education for low back pain and pelvic pain during pregnancy.

Thu, 07/17/2014 - 4:04am
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Manual therapy, exercise, and education for low back pain and pelvic pain during pregnancy.

Am J Obstet Gynecol. 2014 Jun;210(6):592-3

Authors: Licciardone JC, Aryal S

PMID: 24607754 [PubMed - indexed for MEDLINE]

Acute improvement in hemodynamic control after osteopathic manipulative treatment in the third trimester of pregnancy.

Thu, 07/17/2014 - 4:04am
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Acute improvement in hemodynamic control after osteopathic manipulative treatment in the third trimester of pregnancy.

Complement Ther Med. 2013 Dec;21(6):618-26

Authors: Hensel KL, Pacchia CF, Smith ML

Abstract
OBJECTIVES: The physiological changes that occur during pregnancy, including increased blood volume and cardiac output, can affect hemodynamic control, most profoundly with positional changes that affect venous return to the heart. By using Osteopathic Manipulative Treatment (OMT), a body-based modality theorized to affect somatic structures related to nervous and circulatory systems, we hypothesized that OMT acutely improves both autonomic and hemodynamic control during head-up tilt and heel raise in women at 30 weeks gestation.
DESIGN: One hundred subjects were recruited at 30 weeks gestation.
SETTING: The obstetric clinics of UNTHealth in Fort Worth, TX.
INTERVENTION: Subjects were randomized into one of three treatment groups: OMT, placebo ultrasound, or time control. Ninety subjects had complete data (N=25, 31 and 34 in each group respectively).
MAIN OUTCOME MEASURES: Blood pressure and heart rate were recorded during 5 min of head-up tilt followed by 4 min of intermittent heel raising.
RESULTS: No significant differences in blood pressure, heart rate or heart rate variability were observed between groups with tilt before or after treatment (p>0.36), and heart rate variability was not different between treatment groups (p>0.55). However, blood pressure increased significantly (p=0.02) and heart rate decreased (p<0.01) during heel raise after OMT compared to placebo or time control.
CONCLUSIONS: These data suggest that OMT can acutely improve hemodynamic control during engagement of the skeletal muscle pump and this was most likely due to improvement of structural restrictions to venous return.

PMID: 24280470 [PubMed - indexed for MEDLINE]

Evaluation of a novel material, Diomics X-Swab™, for collection of DNA.

Wed, 07/16/2014 - 4:04am
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Evaluation of a novel material, Diomics X-Swab™, for collection of DNA.

Forensic Sci Int Genet. 2014 Jun 24;12C:192-198

Authors: Marshall PL, Stoljarova M, Larue BL, King JL, Budowle B

Abstract
Success of DNA typing is related to the amount of target material recovered from an evidentiary item. Generally, the more DNA that is recovered, the better the chance is of obtaining a typing result that will be robust and reliable. One method of collecting stain materials is by swabbing. Recovery of DNA from a number of commercially available swabs is not an efficient process. The X-Swab™ (Diomics Corporation, La Jolla, CA) is a unique bio-specimen collection material with highly absorptive properties and can be dissolved during certain extraction conditions. Therefore, more DNA may be collected from a substrate and be released from the swab matrix than other swabs. The ability to recover DNA from X-Swab material and success in STR typing were compared with the Copan 4N6FLOQSwab™ (Brescia, Italy), a device which utilizes a proprietary flocked-swab technology to maximize DNA collection and elution efficiency. Both types of swabs were impregnated with known amounts of DNA and body fluids and allowed to air dry. In addition, blood was placed onto glass slides, allowed to dry and collected using both types of swabs. DNA recovery was assessed by DNA quantitation and by STR typing. Results suggested that X-Swab material yielded greater DNA recovery, particularly of low quantity samples (defined as diluted neat samples), compared with the 4N6FLOQSwab. Results also indicated that X-Swab material itself enhances yield of PCR products.

PMID: 25016249 [PubMed - as supplied by publisher]