Organisms Commons^™

Attenuation Of Corneal Myofibroblast Development Through Nanoparticle-Mediated Soluble Transforming Growth Factor-Β Type Ii Receptor (Stgfβrii) Gene Transfer, Ajay Sharma, Jason T. Rodier, Ashish Tandon, Alexander M. Klibanov, Rajiv R. Mohan

Pharmacy Faculty Articles and Research

Purpose: To explore (i) the potential of polyethylenimine (PEI)-DNA nanoparticles as a vector for delivering genes into human corneal fibroblasts, and (ii) whether the nanoparticle-mediated soluble extracellular domain of the transforming growth factor–β type II receptor (sTGFβRII) gene therapy could be used to reduce myofibroblasts and fibrosis in the cornea using an in vitro model.

Methods: PEI-DNA nanoparticles were prepared at a nitrogen-to-phosphate ratio of 30 by mixing linear PEI and a plasmid encoding sTGFβRII conjugated to the fragment crystallizable (Fc) portion of human immunoglobulin. The PEI-DNA polyplex formation was confirmed through gel retardation assay. Human corneal fibroblasts (HCFs) were …

Effects Of In Vivo Hepatic Ischemia-Reperfusion Injury On The Hepatobiliary Disposition Of Rhodamine 123 And Its Metabolites In Isolated Perfused Rat Livers, Ridhi Parasrampuria, Imam H. Shaik, Reza Mehvar

Pharmacy Faculty Articles and Research

Purpose. A few studies have shown that normothermic hepatic ischemia-reperfusion (IR) injury may affect the mRNA and/or protein levels of canalicular transporters P-glycoprotein (P-gp) and multidrug resistance-associated protein 2 (Mrp2). However, the effects of the injury on the functions of these canalicular transporters with respect to the biliary excretion of drugs remain largely unknown. Therefore, the purpose of this study was to investigate the effects of warm hepatic IR on the hepatobiliary disposition of rhodamine 123 (RH-123), a P-gp substrate, and its glucuronidated metabolite (RH-Glu), an Mrp2 substrate, in rats.

Methods. Twenty four or 72 h following a …

The Effects Of 17Α-Ethynylestradiol (Ee2) On Gonadal Development And Differentiation In The Estuarine Killifish, Fundulus Heteroclitus, Ibrahim Chehade

Theses and Dissertations (Comprehensive)

Endocrine disrupting substances (EDSs) comprise a wide variety of chemicals that perturb normal endocrine function including developmental and reproductive processes in vertebrates. The synthetic estrogen 17α-ethynylestradiol (EE₂) is a commonly-used model EDS because of its environmental relevance and its effects on the reproductive endocrine system. Early life-stage exposure of fish to estrogenic EDSs causes effects such as intersex (ovotestes in males) and feminization. This study aims to confirm the period of gonadal differentiation in the estuarine killifish or mummichog (Fundulus heteroclitus) and to determine the sensitivity of gonadal development to EE₂. Artificially-regressed mummichog were spawned, and …

African Elephants Change Gaits When Walking Downhill, Robert H.I. Dale, Robert Warren, Brock Ward, Eric Noble

Scholarship and Professional Work - LAS

Abstract from the International Elephant and Rhino Conservation and Research Symposium, Rotterdam, NL, October 10-14, 2011.

Multistate Mark-Recapture Analysis Reveals No Effect Of Blood Sampling On Survival And Recapture Of Eastern Kingbirds (Tyrannus Tyrannus), Lucas J. Redmond, Michael T. Murphy

Biology Faculty Publications and Presentations

The experimentally supported and prevailing opinion is that blood sampling has few to no long-term effects on survival of birds when conducted properly, and blood sampling has become a vital addition to the toolbox of many ornithologists. However, many of the studies that concluded that blood sampling had negligible effects on birds used approaches that did not account for temporary emigration and probability of capture. To date, the only study to have done so found that blood sampling had a strong negative effect on survival. We conducted a mark–recapture analysis of 8 years of banding and bleeding data on Eastern …

Intermediate Filaments Regulate Tissue Size And Stiffness In The Murine Lens, Douglas S. Fudge, John V. Mccuaig, Shannon Van Stralen, John F. Hess, Huan Wang, Richard T. Mathias, Paul G. Fitzgerald

Biology, Chemistry, and Environmental Sciences Faculty Articles and Research

PURPOSE. To define the contributions of the beaded filament (BF), a lens-specific intermediate filament (IF), to lens morphology and biomechanics.

METHODS. Wild-type and congenic CP49 knockout (KO) mice were compared by using electrophysiological, biomechanical, and morphometric approaches, to determine changes that occurred because of the absence of this cytoskeletal structure.

RESULTS. Electrophysiological assessment established that the fiber cells lacking the lens-specific IFs were indistinguishable from wild-type fiber cells. The CP49 KO mice exhibited lower stiffness, and an unexpected higher resilience than the wildtype lenses. The absence of these filaments resulted in lenses that were smaller, and exhibited a higher ratio …

Targeted Decorin Gene Therapy Delivered With Adeno-Associated Virus Effectively Retards Corneal Neovascularization In Vivo, Rajiv R. Mohan, Jonathan C. K. Tovey, Ajay Sharma, Gregory S. Schultz, John W. Cowden, Ashish Tandon

Pharmacy Faculty Articles and Research

Decorin, small leucine-rich proteoglycan, has been shown to modulate angiogenesis in nonocular tissues. This study tested a hypothesis that tissue-selective targeted decorin gene therapy delivered to the rabbit stroma with adeno-associated virus serotype 5 (AAV5) impedes corneal neovascularization (CNV) in vivo without significant side effects. An established rabbit CNV model was used. Targeted decorin gene therapy in the rabbit stroma was delivered with a single topical AAV5 titer (100 μl; 5x10^12 vg/ml) application onto the stroma for two minutes after removing corneal epithelium. The levels of CNV were examined with stereomicroscopy, H&E staining, lectin, collagen type IV, CD31 immunocytochemistry and …

Polyethylenimine-Conjugated Gold Nanoparticles: Gene Transfer Potential And Low Toxicity In The Cornea, Ajay Sharma, Ashish Tandon, Jonathan C. K. Tovey, Rangan Gupta, J. David Robertson, Jennifer A. Fortune, Alexander M. Klibanov, John W. Cowden, Frank G. Rieger, Rajiv R. Mohan

Pharmacy Faculty Articles and Research

This study examined the gene transfer efficiency and toxicity of 2-kDa polyethylenimine conjugated to gold nanoparticles (PEI2-GNP) in the human cornea in vitro and rabbit cornea in vivo. PEI2-GNP with nitrogen-to-phosphorus (N/P) ratios of up to 180 exhibited significant transgene delivery in the human cornea without altering the viability or phenotype of these cells. Similarly, PEI2-GNP applied to corneal tissues collected after 12 h, 72 h, or 7 days exhibited appreciable gold uptake throughout the rabbit stroma with gradual clearance of GNP over time. Transmission electron microscopy detected GNP in the keratocytes and the extracellular matrix of the rabbit corneas. …

Significant Inhibition Of Corneal Scarring In Vivo With Tissue-Selective, Targeted Aav5 Decorin Gene Therapy, Rajiv R. Mohan, Ashish Tandon, Ajay Sharma, John W. Cowden, Jonathan C. K. Tovey

Pharmacy Faculty Articles and Research

PURPOSE. This study tested a hypothesis that tissue-selective targeted decorin gene therapy delivered to the stroma with adeno-associated virus serotype 5 (AAV5) inhibits corneal fibrosis in vivo without significant side effects.

METHODS. An in vivo rabbit model of corneal fibrosis was used. Targeted decorin gene therapy was delivered to the rabbit cornea by a single topical application of AAV5 (100 L; 6.5 1012 g/mL) onto the bare stroma for 2 minutes. The levels of corneal fibrosis were determined with stereomicroscopy, slit lamp biomicroscopy, -smooth muscle actin ( SMA), fibronectin, and F-actin immunocytochemistry, and/or immunoblotting. CD11b, F4/80 immunocytochemistry, and TUNEL assay …

Efficacious And Safe Tissue-Selective Controlled Gene Therapy Approaches For The Cornea, Rajiv R. Mohan, Sunilima Sinha, Ashish Tandon, Rangan Gupta, Jonathan C. K. Tovey, Ajay Sharma

Pharmacy Faculty Articles and Research

Untargeted and uncontrolled gene delivery is a major cause of gene therapy failure. This study aimed to define efficient and safe tissue-selective targeted gene therapy approaches for delivering genes into keratocytes of the cornea in vivo using a normal or diseased rabbit model. New Zealand White rabbits, adeno-associated virus serotype 5 (AAV5), and a minimally invasive hair-dryer based vector-delivery technique were used. Fifty microliters of AAV5 titer (6.561012 vg/ml) expressing green fluorescent protein gene (GFP) was topically applied onto normal or diseased (fibrotic or neovascularized) rabbit corneas for 2-minutes with a custom vector-delivery technique. Corneal fibrosis and neovascularization in rabbit …

A Comparative Study Of Embedded And Anesthetized Zebrafish In Vivo On Myocardiac Calcium Oscillation And Heart Muscle Contraction, Brian S. Muntean, Christine M. Horvat, James H. Behler, Wissam A. Aboualaiwi, Andromeda M. Nauli, Frederick E. Williams, Surya M. Nauli

Pharmacy Faculty Articles and Research

The zebrafish (Danio rerio) has been used as a model for studying vertebrate development in the cardiovascular system. In order to monitor heart contraction and cytosolic calcium oscillations, fish were either embedded in methylcellulose or anesthetized with tricaine. Using high-resolution differential interference contrast and calcium imaging microscopy, we here show that dopamine and verapamil alter calcium signaling and muscle contraction in anesthetized zebrafish, but not in embedded zebrafish. In anesthetized fish, dopamine increases the amplitude of cytosolic calcium oscillation with a subsequent increase in heart contraction, whereas verapamil decreases the frequency of calcium oscillation and heart rate. Interestingly, verapamil also …

A Survey Of The Management And Development Of Captive African Elephant (Loxodonta Africana) Calves: Birth To Three Months Of Age, Nicole L. Kowalski, Robert H.I. Dale, Christa L. H. Mazur

Scholarship and Professional Work - LAS

We used four surveys to collect information about the birth, physical growth, and behavioral development of 12 African elephant calves born in captivity. The management of the birth process and neonatal care involved a variety of standard procedures. All of the calves were born at night, between 7PM and 7AM. The calves showed a systematic progression in behavioral and physical development, attaining developmental milestones at least a quickly as calves in situ. This study emphasized birth-related events, changes in the ways that calves used their trunks, first instances of behaviors, and interactions of the calves with other, usually adult, elephants. …

Initial Findings On Visual Acuity Thresholds In An African Elephant (Loxodonta Africana), Melissa R. Shyan-Norwalt, Jeff Peterson, Barbara Milankow King, Timothy E. Staggs, Robert H.I. Dale

Scholarship and Professional Work - LAS

There are only a few published examinations of elephant visual acuity. All involved Asian elephants (Elephas maximus) and found visual acuity to be between 8′ and 11′ of arc for a stimulus near the tip of the trunk, equivalent to a 0.50 cm gap, at a distance of about 2 m from the eyes. We predicted that African elephants (Loxodonta africana) would have similarly high visual acuity, necessary to facilitate eye-trunk coordination for feeding, drinking and social interactions. When tested on a discrimination task using Landolt-C stimuli, one African elephant cow demonstrated a visual acuity of …

About Fish, Humane Society Institute For Science And Policy

Aquaculture Collection

Although a number of fish species have been studied in their freshwater life stages, further research on the behavior and habitat requirements of ocean-going fish is required. While fishes have historically been regarded as more “primitive” than other vertebrate groups, Rodriguez et al concluded that several memory and learning systems of bony fishes are noticeably similar to those of reptiles, birds, and mammals. When animals as small and under appreciated as fish display complex mating systems, parental care, and demonstrate the ability to traverse significant distances using olfactory and celestial cues, it is clear that previously established definitions of intelligence …

About Turkeys, The Humane Society Of The United States

Agribusiness Collection

John James Audubon, a well-known bird expert and nature enthusiast, described wild turkeys as birds of great beauty. The history and origin of wild turkeys is uncertain, yet many share Audubon’s sentiment that the wild turkey is “one of the most interesting of the birds indigenous to the United States of America.” Today, wild turkeys can be found throughout the nation. Following the selection of the bald eagle as the American symbol, Benjamin Franklin remarked that the turkey was more “respectable”, and a “true original native”.

About Chickens, The Humane Society Of The United States

Agribusiness Collection

The chicken is the world’s most numerous domesticated bird, with over 52 billion farmed worldwide in 2008, rivaling the dog as the most ubiquitous domestic animal globally. These birds have fascinated scholars and researchers since the dawn of Western civilization, and recent studies are beginning to reveal the depths of their complexity and cognitive ability. According to Andrew F. Fraser, professor of veterinary surgery at Memorial University of Newfoundland, and Donald M. Broom, professor of animal welfare at University of Cambridge: “Those who have studied the behaviour of the domestic fowl in detail…, especially those who have looked at feral …

Gene Delivery In The Equine Cornea: A Novel Therapeutic Strategy, Dylan G. Buss, Ajay Sharma, Elizabeth A. Giuliano, Rajiv R. Mohan

Pharmacy Faculty Articles and Research

Objective—To determine if hybrid adeno-associated virus serotype 2/5 (AAV5) vector can effectively deliver foreign genes into the equine cornea without causing adverse side effects. The aims of this study were to: (i) evaluate efficacy of AAV5 to deliver therapeutic genes into equine corneal fibroblasts (ECFs) using enhanced green fluorescent protein (EGFP) marker gene and (ii) establish the safety of AAV5 vector for equine corneal gene therapy.

Animal Material—Primary ECF cultures were harvested from healthy donor equine corneas. Cultures were maintained at 370C in humidified atmosphere with 5% CO2.

Procedure—AAV5 vector expressing EGFP under control of hybrid cytomegalovirus (CMV) + chicken …

Efficacy And Safety Of Mitomycin C As An Agent To Treat Corneal Scarring In Horses Using An In Vitro Model, Dylan G. Buss, Ajay Sharma, Elizabeth A. Giuliano, Rajiv R. Mohan

Pharmacy Faculty Articles and Research

Objective—Mitomycin C (MMC) is used clinically to treat corneal scarring in human patients. We investigated the safety and efficacy of MMC to treat corneal scarring in horses by examining its effects at the early and late stages of disease using an in-vitro model.

Procedure—An in-vitro model of equine corneal fibroblast (ECF) developed was used. The equine corneal fibroblast or myofibroblast cultures were produced by growing primary ECF in the presence or absence of transforming growth factor beta-1 (TGFβ1) under serum-free conditions. The MMC dose for the equine cornea was defined with dose-dependent trypan blue exclusion and MTT [(3-4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assays …

Isolation And Cultivation Of Equine Corneal Keratocytes, Fibroblasts And Myofibroblasts, Dylan G. Buss, Elizabeth A. Giuliano, Ajay Sharma, Rajiv R. Mohan

Pharmacy Faculty Articles and Research

Objective—To establish an in vitro model for the investigation of equine corneal wound healing. To accomplish this goal, a protocol to isolate and culture equine corneal keratocytes, fibroblasts and myofibroblasts was developed.

Animal material—Equine corneal buttons were aseptically harvested from healthy research horses undergoing humane euthanasia for reasons unrelated to this study. Slit-lamp biomicroscopy was performed prior to euthanasia by a board-certified veterinary ophthalmologist to ensure that all samples were harvested from horses free of anterior segment disease.

Procedure—Equine corneal stroma was isolated using mechanical techniques and stromal subsections were then cultured. Customized media at different culture conditions was used …

Vector Delivery Technique Affects Gene Transfer In The Cornea In Vivo, Rajiv R. Mohan, Ajay Sharma, Tyler C. Cebulko, Ashish Tandon

Pharmacy Faculty Articles and Research

Purpose: This study tested whether controlled drying of the cornea increases vector absorption in mouse and rabbit corneas in vivo and human cornea ex vivo, and studied the effects of corneal drying on gene transfer, structure and inflammatory reaction in the mouse cornea in vivo.

Methods: Female C57 black mice and New Zealand White rabbits were used for in vivo studies. Donor human corneas were used for ex vivo experiments. A hair dryer was used for drying the corneas after removing corneal epithelium by gentle scraping. The corneas received no, once, twice, thrice, or five times warm air for …

Plasma Pharmacokinetics And Tissue Disposition Of Novel Dextran- Methylprednisolone Conjugates With Peptide Linkers In Rats, Suman Penugonda, Hitesh K. Agarwal, Keykavous Parang, Reza Mehvar

Pharmacy Faculty Articles and Research

The plasma and tissue disposition of two novel dextran prodrugs of methylprednisolone (MP) containing one (DMP-1) or five (DMP-5) amino acids as linkers were studied in rats. Single 5-mg/kg doses (MP equivalent) of each prodrug or MP were administered intravenously, and blood and tissue samples were collected. Prodrug and drug concentrations were quantitated using HPLC, and noncompartmental pharmacokinetic parameters were estimated. Whereas conjugation of MP with dextran in both prodrugs substantially decreased the clearance of the drug by ∼200-fold, the accumulations of the drug in the liver, spleen, and kidneys were significantly increased by conjugation. However, the extent of accumulation …

Localization Of Angiotensin Converting Enzyme In Rabbit Cornea And Its Role In Controlling Corneal Angiogenesis In Vivo, Ajay Sharma, Daniel I. Bettis, John W. Cowden, Rajiv R. Mohan

Pharmacy Faculty Articles and Research

Purpose: The renin angiotensin system (RAS) has been shown to modulate vascular endothelial growth factor and angiogenesis. In this study we investigated (i) the existence of the RAS components angiotensin converting enzyme (ACE) and angiotensin II receptors (AT1 and AT2) in the rabbit cornea using in vitro and ex vivo models and (ii) the effect of enalapril, an ACE inhibitor, to inhibit angiogenesis in rabbit cornea in vivo.

Methods: New Zealand White rabbits were used. Cultured corneal fibroblasts and corneal epithelial cells were used for RNA isolation and cDNA preparation using standard molecular biology techniques. PCR was performed to …

Aav Serotype Influences Gene Transfer In Corneal Stroma In Vivo, Ajay Sharma, Jonathan C. K. Tovey, Arkasubhra Ghosh, Rajiv R. Mohan

Pharmacy Faculty Articles and Research

This study evaluated the cellular tropism and relative transduction efficiency of three AAV serotypes, AAV6, AAV8 and AAV9, for corneal gene delivery using mouse cornea in vivo and donor human cornea ex vivo. The AAV6, AAV8 and AAV9 serotypes having AAV2 plasmid encoding for alkaline phosphatase (AP) gene were generated by transfecting HEK293 cell line with pHelper, pARAP4 and pRep/Cap plasmids. Viral vectors (109 vg/μl) were topically applied onto mouse cornea in vivo and human cornea ex vivo after removing the epithelium. Human corneas were processed for transgene delivery at day 5 after viral vector application. Mouse corneas were harvested …

Different Training Volumes Yield Equivalent Increases In Bmd, R. A. Pierce, L. C. Lee, C. P. Ahles, S. M. Shdo, S. V. Jaque, Ken D. Sumida

Biology, Chemistry, and Environmental Sciences Faculty Articles and Research

The purpose of this study was to determine if an exercise threshold existed in stimulating an elevation in bone mineral density (BMD), via resistance training, during the growth period in male rats. 27 male rats were randomly divided into Control (Con, n = 9), 3 ladder climb resistance trained group (3LC, n = 9), and 6 ladder climb resistance trained group (6LC, n = 9). The 3LC and 6LC groups were conditioned to climb a vertical ladder with weights appended to their tail 3 days/wk for a total of 6 wks, but the 6LC group performed significantly more work than …

Equal Bmd After Daily Or Triweekly Exercise In Growing Rats, B. D. Kayser, J. K. Godfrey, R. M. Cunningham, R. A. Pierce, S. V. Jaque, Ken D. Sumida

Biology, Chemistry, and Environmental Sciences Faculty Articles and Research

The purpose of this study was to examine the efficacy of continuous resistance training (3 days/wk) compared to interrupted resistance training where 20-24 h separated an exercise bout (i.e. 6 days/wk) for enhancing bone mineral density (BMD) in growing male rats. The total volume of work performed per week between the two resistance training programs was equivalent by design. Young male rats were randomly divided into Control (Con, n = 9), 3 days/wk resistance trained group (RT3, n = 9), and 6 days/wk resistance trained group (RT6, n = 9). The RT3 and RT6 groups were conditioned to climb a …

The Transition Zone: Impact Of Riverbanks On Emergent Dragonfly Nymphs. Implications For Riverbank Restoration And Management, Kirsten Hope Martin

Antioch University Dissertations & Theses

The use of riprap in the restoration and stabilization of riverine landscapes is an issue of concern for many ecologists. While current methods of bank stabilization, especially those involving the placement of rocks (riprap) along the waterline, are effective in controlling erosion their presence changes habitat components (slope, substrate composition, near-shore river velocity) at the river-land interface. The additional impacts of river current, water temperature, soil composition, slope, and water level fluctuation, may further imperil emerging nymphs. The purpose of this research is to document the effects of riprap, location (upriver or downriver of hydroelectric intake/outtake facilities), water level fluctuation, …

Female Humpback Whale (Megaptera Novaeangliae) Reproductive Class And Male-Female Interactions During The Breeding Season, Meagan E. Jones

Antioch University Dissertations & Theses

This study examined whether female reproductive class (i.e., presence or absence of a calf) in humpback whales (Megaptera novaeangliae) affects female behavior and male-female interactions on the Hawaiian breeding grounds. From 2003-2008, 295 female-calf groups (with or without a male escort) and 256 female no-calf groups (with a male) were observed. Forty-one female no-calf groups were confirmed male-female pairs by genetic sexing or photo-identification. Focal follows of 36 male-female pairs and 50 female-calf groups (29 escorted and 21 unescorted female-calf groups) were analyzed for time budget and movement estimates. Relative fluke size (fluke photogrammetry, n=30) and sighting histories …

Trichostatin A Inhibits Corneal Haze In Vitro And In Vivo, Ajay Sharma, Maneesh M. Mehan, Sunilima Sinha, John W. Cowden, Rajiv R. Mohan

Pharmacy Faculty Articles and Research

PURPOSE. Trichostatin A (TSA), a histone deacetylase inhibitor, has been shown to suppress TGF- –induced fibrogenesis in many nonocular tissues. The authors evaluated TSA cytotoxicity and its antifibrogenic activity on TGF- –driven fibrosis in the cornea with the use of in vitro and in vivo models.

METHODS. Human corneal fibroblasts (HSFs) were used for in vitro studies, and New Zealand White rabbits were used for in vivo studies. Haze in the rabbit cornea was produced with photorefractive keratectomy (PRK) using excimer laser. Trypan blue exclusion and MTT assays evaluated TSA cytotoxicity to the cornea. Density of haze in the rabbit …

Interrupted Resistance Training And Bmd In Growing Rats, J. K. Godfrey, B. D. Kayser, G. V. Gomez, Jason Bennett, S. V. Jaque, Ken D. Sumida

Biology, Chemistry, and Environmental Sciences Faculty Articles and Research

A resistance training program, where the exercise was uninterrupted (UT, i.e. continuous repetitions) was compared against another resistance training program where the exercise was interrupted (IT, i.e. 2 exercise sessions during a training day) for enhancing bone modeling and bone mineral density (BMD) in maturating animals. The total volume of work performed between the two resistance training programs was equivalent by design. Young male rats (similar to 8 weeks old) were randomly divided into Control (Con, n = 8), UT (n = 8) and IT (n = 7) resistance trained groups. The UT and IT groups were conditioned to climb …

Liquid Chromatography-Tandem Mass Spectrometry For The Determination Of Methylprednisolone In Rat Plasma And Liver After Intravenous Administration Of Its Liver-Targeted Dextran Prodrug, Shuang-Qing Zhang, Helen R. Thorsheim, Suman Penugonda, Venkateswaran C. Pillai, Quentin R. Smith, Reza Mehvar

Pharmacy Faculty Articles and Research

A specific and sensitive liquid chromatography (LC)-tandem mass spectrometric method for quantitative determination of methylprednisolone (MP) in rat plasma and liver was developed and validated using triamcinolone acetonide as an internal standard. Liquid-liquid extraction using tert-butyl methyl ether was used to extract the drug and the internal standard from plasma and liver. The separation of MP was performed on a C(18) column with a mobile phase of acetonitrile:0.5% formic acid aqueous solution (85:15, v/v) over 4 min. The assay was based on the selected reaction monitoring transitions at m/z 375 -> 161 for MP in plasma, 375 -> 357 for …