Veera Rajaratnam - Academia.edu (original) (raw)

Papers by Veera Rajaratnam

with green tea extract: a pilot randomized

Investigative Ophthalmology & Visual Science, 2004

Investigative Ophthalmology & Visual Science, 2003

Gynecologic and Obstetric Investigation, 2014

Background/Aims: To investigate the inhibitory effect of green tea extract, epigallocatechin gall... more Background/Aims: To investigate the inhibitory effect of green tea extract, epigallocatechin gallate (EGCG), on wild-type human leiomyoma (WT-HuLM) cells and its potential action via catechol-o-methyltransferase (COMT) activity. Methods: Cell proliferation of WT-HuLM and COMT gene-silenced HuLM (COMT-shRNA-HuLM) cells treated with 0 or 100 µM EGCG for 7 days was measured using the MTT method. Total RNA and protein were extracted from cells treated with 0 or 100 µM of EGCG for 48 h. Gene expression profiling was performed using Human Signal Transduction PathwayFinder. Proliferation cell nuclear antigen (PCNA), cyclin-dependent kinase 4 (Cdk4) and COMT protein levels were detected by Western blot analyses. COMT enzyme activity was evaluated by HPLC. Results: EGCG-treated WT-HuLM cells showed significantly decreased COMT expression (p < 0.001) and enzyme activity (p < 0.05) compared to untreated WT-HuLM cells, while COMT-shRNA-HuLM cells showed no significant change. At 100 μM of...

Molecular Carcinogenesis, 2008

Soy protein is known to have breast tumor suppressing activity. The expression of peripheral benz... more Soy protein is known to have breast tumor suppressing activity. The expression of peripheral benzodiazepine receptors (PBRs), currently renamed as translocator protein (TSPO) and their associated functions, such as nuclear cholesterol uptake and content also have been shown to be increased in breast cancer. Here we investigated whether the breast tumor suppressing effects of soy protein is mediated by down-regulation of PBR expression and function. Breast tumors were induced by gavage administration of a single dose (80 mg/kg) of dimethylbenz[a]anthracene (DMBA) into 50-d old female Sprague Dawley rats, maintained on a standard AIN-76A diet containing either casein or soy protein. Approximately 120 d following DMBA administration, the animals were sacrificed. All tumors were detected by palpation and at autopsy biopsy specimens were taken for histological grading. The ligand binding capacity, expression, and protein levels of PBRs, their nuclear localization and function, such as nuclear cholesterol uptake and content, were significantly increased in the tumors. However, replacement of casein by soy protein in the diet caused a significant decrease in all of these parameters. These data suggest that soy protein inhibits breast tumor development by decreasing the expression of the tumor-promoting gene, which encodes PBRs.

Journal of Biochemical and Molecular Toxicology, 2009

2-Choloroethyl Ethyl Sulfide (CEES) exposure causes inflammatory lung diseases, including acute r... more 2-Choloroethyl Ethyl Sulfide (CEES) exposure causes inflammatory lung diseases, including acute respiratory distress syndrome (ARDS) and pulmonary fibrosis. This may be associated with oxidative stress, which has been implicated in the desensitization of beta-adrenergic receptors (β-ARs). The objective of this study was to investigate whether lung injury induced by intratracheal CEES exposure (2 mg/kg body weight) causes desensitization of β-ARs. The animals were sacrificed after 7 days and lungs were removed. Lung injury was established by measuring the leakage of iodinated-bovine serum albumin ([ 125 I]-BSA) into lung tissue. Receptor-binding characteristics were determined by measuring the binding of [ 3 H] dihydroalprenolol ([ 3 H] DHA) (0.5-24 nM) to membrane fraction in the presence and absence of DL-propranolol (10 μM). Both high-and low-affinity β-ARs were identified in the lung. Binding capacity was significantly higher in low-affinity site in both control and experimental groups. Although CEES exposure did not change K D and B max at the high-affinity site, it significantly decreased both K D and B max at low affinity sites. A 20% decrease in β 2-AR mRNA level and a 60% decrease in membrane protein levels were observed in the experimental group. Furthermore, there was significantly less stimulation of adenylate cyclase activity by both cholera toxin and isoproterenol in the experimental group in comparison to the control group. Treatment of lungs with 3-isobutyl-1-methylxanthine (IBMX), an inhibitor of phosphodiesterase (PDE) could not abolish the difference between the control group and the experimental group on the stimulation of the adenylate cyclase activity. Thus, our study indicates that CEES-induced lung injury is associated with desensitization of β 2-AR.

Journal of Biochemical and Molecular Toxicology, 2005

Exposure to mustard gas causes inflammatory lung diseases, including acute respiratory distress s... more Exposure to mustard gas causes inflammatory lung diseases, including acute respiratory distress syndrome (ARDS). A defect in the lung surfactant system has been implicated as a cause of ARDS. A major component of lung surfactant is dipalmitoyl phosphatidylcholine (DPPC) and the major pathway for its synthesis is the cytidine diphosphocholine (CDP-choline) pathway. It is not known whether the ARDS induced by mustard gas is mediated by its direct effects on some of the enzymes in the CDPcholine pathway. In the present study, we investigated whether mustard gas exposure modulates the activity of cholinephosphotransferase (CPT), the terminal enzyme by CDP-choline pathway. Adult guinea pigs were intratracheally infused with single doses of 2chloroethyl ethyl sulfide (CEES) (0.5 mg/kg b.wt. in ethanol). Control animals were injected with vehicles only. The animals were sacrificed at different time and the lungs were removed after perfusion with physiological saline. CPT activity increased steadily up to 4 h and then decreased at 6 h and stabilized at 7 days in both mitochondria and microsomes. To determine the dose-dependent effect of CEES on CPT activity, we varied the doses of CEES (0.5-6.0 mg/kg b.wt.) and sacrificed the animals at 1 h and 4 h. CPT activity showed a dose-dependent increase of up to 2.0 mg/kg b.wt. of CEES in both mitochondria and microsomes, and then decreased at 4.0 mg/kg b.wt. For further studies, we used a fixed single dose of CEES (2.0 mg/kg b.wt.) and fixed exposure time (7 days). Lung injury was determined by measuring the leakage of iodinated-bovine serum albumin into lung tissue and expressed as the permeability index. CEES exposure (2.0 mg/kg b.wt. for 7 days) caused a significant decrease

Journal of Biochemical and Molecular Toxicology, 2006

Mustard gas exposure causes inflammatory lung diseases. Many inflammatory lung diseases are assoc... more Mustard gas exposure causes inflammatory lung diseases. Many inflammatory lung diseases are associated with oxidative stress. Reactive oxygen species (ROS) are involved in the maintenance of physiological functions. In tissues, it is therefore essential to maintain a steady-state level of antioxidant activity to allow both for the physiological functions of ROS to proceed and at the same time preventing tissue damage. We have recently reported that mustard gas exposure decreases the overall activity of superoxide dismutase (SOD). In the present study, we investigated the effects of mustard gas on each of the three isozymes: SOD-1 (Cu/Zn), SOD-2 (Mn), and SOD-3 (extracellular). Adult guinea pigs were intratracheally injected single doses of 2-chloroethyl ethyl sulfide (CEES) (2 mg/kg body weight) in ethanol. Control animals were injected with vehicle in the same way. The animals were sacrificed after 7 days, and lungs were removed after perfusion with physiological saline. Lung injury was established by measuring the leakage of iodinated-BSA into lung tissue. Mustard gas exposure caused a significant increase in the activity of SOD-1 (35%). However, the SOD-3 activity which is the predominant type in lung was significantly decreased (62%), whereas no change was observed in SOD-2 activity. Thus the decrease in the total activity of SOD was primarily due to the SOD-3 isozyme. Northern blot analysis indicated 3.5-fold increased expression of SOD-1 in mustard gas exposed lung, but no significant change in the expression of SOD-2 and SOD-3 was observed. Mustard gas exposure did not cause mutation in the coding region of SOD-1 gene while causing modulation in expression levels. The protein levels of SOD-1, SOD-2, and SOD-3 were not altered significantly in the mustard gas exposed lung. Our results indicate that

Investigative Opthalmology & Visual Science, 2006

To characterize the angiostatic effect of penetrating ocular injury and to begin to explore its m... more To characterize the angiostatic effect of penetrating ocular injury and to begin to explore its mechanism, with an emphasis on the role of pigment epithelium-derived factor (PEDF). METHODS. Using the rat model of oxygen-induced retinopathy (OIR), single or multiple dry needle injuries were made, penetrating the globe of one eye; the opposite eye served as a control. Eyes were harvested from rats killed 1, 3, and 6 days after injury, and retinas were dissected and processed for assessment of neovascularization and microglial activation or were processed for genetic and proteomic analysis. Temporal and spatial expression patterns of PEDF were analyzed by in situ hybridization. RESULTS. Penetrating ocular injury resulted in a 30% decrease in neovascular area in the retinas of OIR rats. At day 1 after injury, needle insertion caused a 4.1-fold increase in retinal PEDF mRNA and a 1.5-fold increase in retinal PEDF protein. Vitreous PEDF protein increased 3.4-fold in injured eyes compared with noninjured eyes. In situ hybridization showed an increase in PEDF mRNA in areas surrounding the puncture site. Concentrated vitreous protein from injured eyes caused a 60% decrease in retinal neovascularization when injected into the vitreous cavity of OIR rats. Preincubation of vitreous samples with anti-PEDF partially abolished this efficacy. CONCLUSIONS. The pattern of angiostasis resulting from penetrating ocular injury is consistent with the release of an endogenous antiangiogenic factor from the wound site. Preliminary studies show a possible role for PEDF in this effect. Further characterization of this role and the identification of other factors may lead to new therapeutic strategies for angiogenic eye conditions.

Investigative Opthalmology & Visual Science, 2003

PURPOSE Restructuring of extracellular matrix at actively extending blood vessel tips involves se... more PURPOSE Restructuring of extracellular matrix at actively extending blood vessel tips involves secretion of plasminogen activator (PA). Findings in earlier studies conducted in the authors' laboratory have suggested that angiostatic steroids suppress the PA activity essential for the invasive aspect of angiogenesis by increasing synthesis of plasminogen activator inhibitor (PAI)-1. This experiment was designed to test the effect of administration of exogenous PAI-1 on retinal neovascularization (NV) in an animal model of retinopathy of prematurity (ROP). METHODS At birth, Sprague-Dawley rats were placed into incubators and exposed to an atmosphere alternating between 50% and 10% O 2 every 24 hours. After 14 days, the animals were removed to room air, at which time each received a single intravitreal injection of 5 L of buffer vehicle or one of five doses of PAI-1, ranging from 3.0 g/mL to 2.0 mg/mL. Animals were killed 6 days later, and retinal NV was assessed using adenosine diphosphatase (ADPase) histochemical staining. RESULTS Retinal neovascularization decreased with increasing PAI-1 dosage. The most effective dose tested (2.0 mg/mL) caused a 52% reduction in retinal NV relative to vehicle (P Ͻ 0.005). Normal vasculogenesis, as determined by measuring retinal vascular area, was unaffected. CONCLUSIONS PAI-1 inhibits pathologic angiogenesis without adversely affecting normal vasculogenesis, an attractive feature for ROP therapies. Moreover, PAI's relationship to matrix metalloproteinases, which are also implicated in angiogenesis, suggests that the proteolytic aspect of the process may provide additional downstream therapeutic targets. (Invest Ophthal

International Journal of Women's Health, 2013

Background: Uterine fibroids (UFs, also known as leiomyoma) affect 70% of reproductive-age women.... more Background: Uterine fibroids (UFs, also known as leiomyoma) affect 70% of reproductive-age women. Imposing a major burden on health-related quality-of-life (HRQL) of premenopausal women, UF is a public health concern. There are no effective medicinal treatment options currently available for women with symptomatic UF. Objectives: To evaluate the efficacy and safety of green tea extract (epigallocatechin gallate [EGCG]) on UF burden and quality of life in women with symptomatic UF, in a double-blinded, placebo-controlled randomized clinical trial. Methods: A total of 39 reproductive-age women (age 18-50 years, day 3 serum folliclestimulating hormone ,10 mIU/mL) with symptomatic UF were recruited for this study. All subjects had at least one fibroid lesion 2 cm 3 or larger, as confirmed by transvaginal ultrasonography. The subjects were randomized to oral daily treatment with either 800 mg of green tea extract (45% EGCG) or placebo (800 mg of brown rice) for 4 months, and UF volumes were measured at the end, also by transvaginal ultrasonography. The fibroid-specific symptom severity and HRQL of these UF patients were scored at each monthly visit, using the symptom severity and quality-of-life questionnaires. Student's t-test was used to evaluate statistical significance of treatment effect between the two groups. Results: Of the final 39 women recruited for the study, 33 were compliant and completed all five visits of the study. In the placebo group (n = 11), fibroid volume increased (24.3%) over the study period; however, patients randomized to green tea extract (n = 22, 800 mg/day) treatment showed significant reduction (32.6%, P = 0.0001) in total UF volume. In addition, EGCG treatment significantly reduced fibroid-specific symptom severity (32.4%, P = 0.0001) and induced significant improvement in HRQL (18.53%, P = 0.01) compared to the placebo group. Anemia also significantly improved by 0.7 g/dL (P = 0.02) in the EGCG treatment group, while average blood loss significantly decreased from 71 mL/month to 45 mL/month (P = 0.001). No adverse effects, endometrial hyperplasia, or other endometrial pathology were observed in either group. Conclusion: EGCG shows promise as a safe and effective therapeutic agent for women with symptomatic UFs. Such a simple, inexpensive, and orally administered therapy can improve women's health globally.

Injury Extra, 2007

, and whose medical notes were available were included in the audit. Results: Sixty-eight patient... more , and whose medical notes were available were included in the audit. Results: Sixty-eight patients were available for analysis. They included 44 (65%) male and 24 (35%) female with mean age of 36 (2-101). There were varying grades of injury: 24% Gustillo and Anderson Grade I, 39% Grade II, 16% Grade IIIa, 18% IIIb and 3% IIIc. Only 3% of cases had documented evidence of appropriate wound care in A&E. Ninety-three percent of patients received IV antibiotics given at a mean time from injury of 2 h 37 min. Of the 44% of cases referred to plastics, only 13% (4/30) of referrals were made before the initial procedure. The mean time from injury to initial operative procedure was 9 h 34 min (median 5 h 22 min). Only 23% of cases were carried out with a consultant present. Ten of 62 cases (six lost to follow-up) were complicated by infection, 2 (3%) of which were deep. Two cases of compartment syndrome occurred requiring fasciotomy. All fractures united with a mean time to union of 39 weeks. The most striking problem was the poor quality of note keeping and hence difficulty in obtaining accurate data. Conclusions: The results show that in a number of crucial areas the guidelines are not adhered to. However, the complication rate in terms of nonunion and deep infection is respectable compared to current literature. One cannot read too much into this due to the problems with data collection. The department should examine its current protocol and perform a prospective audit thereby obtaining an accurate picture of its results. In the meantime emergent debridement of soft tissues, fracture stabilisation and early administration of intravenous antibiotics should be supported.

Human Reproduction, 2013

Is targeted adenovirus vector, Ad-SSTR-RGD-TK (Adenovirus-human somatostatin receptor subtype 2-a... more Is targeted adenovirus vector, Ad-SSTR-RGD-TK (Adenovirus-human somatostatin receptor subtype 2-arginine, glycine and aspartate-thymidine kinase), given in combination with ganciclovir (GCV) against immortalized human leiomyoma cells (HuLM) a potential therapy for uterine fibroids? summary answer: Ad-SSTR-RGD-TK/GCV, a targeted adenovirus, effectively reduces cell growth in HuLM cells and to a significantly greater extent than in human uterine smooth muscle cells (UtSM). what is known already: Uterine fibroids (leiomyomas), a major cause of morbidity and the most common indication for hysterectomy in premenopausal women, are well-defined tumors, making gene therapy a suitable and potentially effective non-surgical approach for treatment. Transduction of uterine fibroid cells with adenoviral vectors such as Ad-TK/GCV (herpes simplex virus thymidine kinase gene) decreases cell proliferation. study design, size, duration: An in vitro cell culture method was set up to compare and test the efficacy of a modified adenovirus vector with different multiplicities of infection in two human immortalized cell lines for 5 days. participants/materials, setting, methods: Immortalized human leiomyoma cells and human uterine smooth muscle cells were infected with different multiplicities of infection (MOI) (5-100 plaque-forming units (pfu)/cell) of a modified Ad-SSTR-RGD-TK vector and subsequently treated with GCV. For comparison, HuLM and UtSM cells were transfected with Ad-TK/GCV and Ad-LacZ/GCV. Cell proliferation was measured using the CyQuant assay in both cell types. Additionally, western blotting was used to assess the expression of proteins responsible for regulating proliferation and apoptosis in the cells. main results and the role of chance: Transduction of HuLM cells with Ad-SSTR-RGD-TK/GCV at 5, 10, 50 and 100 pfu/cell decreased cell proliferation by 28, 33, 45, and 84%, respectively (P , 0.05) compared with untransfected cells, whereas cell proliferation in UtSM cells transfected with the same four MOIs of Ad-SSTR-RGD-TK/GCV compared with that of untransfected cells was decreased only by 8, 23, 25, and 28%, respectively (P , 0.01). Western blot analysis showed that, in comparison with the untargeted vector Ad-TK, Ad-SSTR-RGD-TK/GCV more effectively reduced expression of proteins that regulate the cell cycle (Cyclin D1) and proliferation (PCNA, Proliferating Cell Nuclear Antigen), and it induced expression of the apoptotic protein BAX, in HuLM cells. limitations, reasons for caution: Results from this study need to be replicated in an appropriate animal model before testing this adenoviral vector in a human trial. wider implications of the findings: Effective targeting of gene therapy to leiomyoma cells enhances its potential as a noninvasive treatment of uterine fibroids.

Gynecologic and Obstetric Investigation, 2013

Postoperative abdominal/pelvic peritoneal adhesions are a major source of morbidity (bowel obstru... more Postoperative abdominal/pelvic peritoneal adhesions are a major source of morbidity (bowel obstruction, infertility, ectopic gestation as well as chronic pelvic pain) in women. In this study, we screened various transduction and transcription modifications of adenovirus (Ad) to identify those that support maximal Ad-mediated gene delivery to human adhesion fibroblasts, which in turn would enhance the efficacy of this novel treatment/preventative strategy for postoperative adhesions. We transduced primary cultures of human peritoneal adhesion fibroblasts with fiber-modified Ad vectors Ad5-RGD-luc, Ad5-Sigma-luc, Ad5/3-luc and Ad5-CAV2-luc as well as transcriptional targeting viruses Ad5-survivin-luc, Ad5-heparanase-luc, Ad5-mesothelin (MSLN)-CRAd-luc and Ad5-secretory leukoprotease inhibitor (SLPI)-luc, and compared their activity to wild-type Ad5-luc. At 48 h, luciferase activity was measured and normalized to the total protein content in the cells. Among the fiber-modified Ad vecto...

Fertility and Sterility, 2010

OBJECTIVE: Mullerian and male urogenital anomalies share common genetic, hormonal and environment... more OBJECTIVE: Mullerian and male urogenital anomalies share common genetic, hormonal and environmental etiologic factors. The purpose of this study is to quantify the risk of urogenital anomalies in male relatives of women with Mullerian anomalies. DESIGN: Kinship analysis. MATERIALS AND METHODS: Patients were identified by ICD codes from the largest hospitals in the state of Utah (1994 to 2009). We included all patients who were coded for doubling of the uterus or Mullerian anomalies. Controls for the probands were matched based on birth year and sex (1:5). All records were subsequently matched to the Utah population data base (UPDB) to obtain pedigrees with inclusion of data regarding male urogenital anomalies and to perform kinship analysis. The prevalence of the following urogenital anomalies (undescended testis, hypospadias, epispadias, anomalies of the penis, or other male genital anomalies) in all male relatives of the probands was compared to controls. The Kinship analysis software kingless was used to compute the relative risks for urogenital anomalies among male relatives of probands with female Mullerian anomalies. RESULTS: The probands comprised 2939 women with Mullerian anomalies. Males with urogenital anomalies identified through ICD codes were 10176 patients. First degree (children, parent) male relatives of women with Mullerian anomalies had a RR of urogenital anomalies of 1.43 (1.09-1.89). This risk was largely limited to children of the probands (RR¼1.47, 1.1-1.97). No parents of the probands or controls were affected. Relative risks for the other kinship classes were not significantly elevated. The prevalence of anomalies in male children of women with Mullerian anomalies were as follows: undescended testis (53%),hypospadias (36%), epispadias (0%), anomalies of the penis (29%) and other male genital anomalies (1.5%). CONCLUSION: Urogenital anomalies appear to be increased in male children of women with Mullerian anomalies. Undescended testis and hypospadias are the most common anomalies. Supported by: Departmental. OBJECTIVE: Uterine fibroids are the most common benign tumors of women. Fibroids are more common in African American (AA), and the prevalence increases with age, but little is known about other races. We examined the effects of age, race, BMI, and hormonal markers on fibroid prevalence in a large, multi-ethnic population. DESIGN: Cross-sectional.

Experimental Eye Research, 2004

Retinal capillary quiescence is regulated by a delicate balance between proangiogenic and anti-an... more Retinal capillary quiescence is regulated by a delicate balance between proangiogenic and anti-angiogenic factors. Pathological angiogenesis is the result of a shift in this balance towards proangiogenic influences. Pathological angiogenesis is produced in a rat model of oxygen-induced retinopathy (OIR) by exposing newborn rat pups to alternating periods of hyperoxia and hypoxia. Based upon previous work, two similar exposure paradigms were investigated and compared, exposure of rat pups to alternating periods of 45 and 12$5% oxygen, and to alternating periods of 40 and 15% oxygen. The resulting retinal pathology was assessed by measurement of retinal clock hours with pathological blood vessel growth and the percentage of the retina that is avascular. The 45 and 12$5% exposure produced significantly greater incidence and severity of pathology than the 40 and 15% protocol. To explain the difference in pathology between these two very similar exposure protocols, retinal levels of proangiogenic vascular endothelial growth factor (VEGF) and VEGF receptor 2 (VEGFR2) and anti-angiogenic pigment epithelium-derived factor (PEDF) were measured by ELISA and western blot analysis at 0, 2, and 6 days postexposure. In whole retinal lysates, there were no significant differences in VEGFR2 and PEDF levels. However, VEGF levels were approximately 48 and 78% higher on post-oxygen exposure day 0 and 2, respectively, in the group treated with alternating periods of 45 and 12$5% oxygen compared to the group treated with alternating periods of 40 and 15% oxygen. There was no significant difference in VEGF levels between these two groups on day 6 post-exposure. Therefore, the difference in pathology observed between these two experimental paradigms is associated with differences in whole retinal VEGF levels, but not changes in whole retinal VEGFR2 or PEDF levels. The results of this study suggest the existence of a threshold in the rat model of OIR, such that a small change in blood oxygen profile triggers a disproportionate increase in subsequent neovascularization, which is accompanied by more dramatic changes of retinal VEGF level than VEGFR2 or PEDF level. If a similar threshold exists for humans, it could explain why some oxygen-treated premature infants develop retinopathy and others do not, despite similar gestational ages, birth weights and clinical courses.

American Journal of Obstetrics and Gynecology, 2010

Objective-Investigate the effect of epigallocatechin gallate (EGCG), on rat leiomyoma (ELT3) cell... more Objective-Investigate the effect of epigallocatechin gallate (EGCG), on rat leiomyoma (ELT3) cells in vitro and in nude mice model. Study Design-ELT3 cells were treated with various concentrations of EGCG. Cell proliferation, PCNA and Cdk4 protein levels were evaluated. ELT3 cells were inoculated subcutaneously in female athymic nude mice. Animals were fed 1.25mg EGCG (in drinking water)/mouse/day. Tumors were collected and evaluated at 4 and 8 weeks post-treatment. Results-Inhibitory effect of EGCG (200 μM) on ELT3 cells was observed after 24 h treatment (p<0.05). At ≥50μM, EGCG significantly decreased PCNA and Cdk4 protein levels (p<0.05). In vivo, EGCG treatment dramatically reduced the volume and weight of tumors at 4 and 8 weeks posttreatment (p<0.05). The PCNA and Cdk4 protein levels were significantly reduced in EGCG treated group (p<0.05). Conclusion-EGCG effectively inhibits the proliferation and induce apoptosis in rat ELT3 uterine leiomyoma cells in vitro and in vivo.

Experimental Eye Research, 2004

The pathogenesis of retinopathy of prematurity involves dysregulated angiogenesis resulting in pr... more The pathogenesis of retinopathy of prematurity involves dysregulated angiogenesis resulting in pre-retinal growth of new vessels. Inhibition of tyrosine kinase-dependent pro-angiogenic signals may provide a rational therapeutic approach to the reduction of pre-retinal neovascularization. Vascular endothelial growth factor stimulates endothelial cell mitogenesis, differentiation and migration, by binding and activating the receptor tyrosine kinases vascular endothelial growth factor receptor-1 and vascular endothelial growth factor receptor-2. One of the vascular endothelial growth factor receptor substrates implicated in vascular endothelial growth factor signal transduction is c-Src. The ability of herbimycin A, a c-Src-selective tyrosine kinase inhibitor, to inhibit vascular endothelial growth factor-induced bovine retinal microvascular endothelial cell proliferation and tube formation was investigated. The ability of the compound to inhibit pathologic angiogenesis was tested in a rat model of retinopathy of prematurity. Exposure of neonatal rats to oxygen concentrations cycling between 10 and 50% induced severe pre-retinal neovascularization in all rats. Some of the eyes of these variable oxygen-exposed rats were herbimycin A-injected or vehicle-injected 1 or 3 days post-oxygen exposure while some eyes were non-injected. All rats were sacrificed for assessment 6 days post-exposure. Herbimycin A inhibited both vascular endothelial growth factor-induced bovine retinal microvascular endothelial cell proliferation and capillary tube formation in a dose-dependent manner. Injection of herbimycin A into oxygen-treated rats 1 day post-oxygen exposure produced a 63% decrease in pre-retinal neovascularization relative to vehicle (P = 0.0029). There was a 41% decrease in pre-retinal neovascularization in herbimycin-injected eyes relative to vehicle-injected eyes 3 days post-oxygen (P = 0.031). Pre-retinal neovascularization was reduced in vehicle-injected eyes relative to non-injected eyes at both injection times. There were no significant differences in retinal vascular area between any of the experimental groups. Based on the results of this study, herbimycin A inhibits endothelial cell proliferation and tube formation at non-toxic concentrations and reduces pre-retinal neovascularization in a rat model of retinopathy of prematurity. Reduction of angiogenesis by the inhibition of tyrosine kinase activity may be a viable route to the development of effective chemotherapies applicable to eye disease.

with green tea extract: a pilot randomized

Investigative Ophthalmology & Visual Science, 2004

Investigative Ophthalmology & Visual Science, 2003

Gynecologic and Obstetric Investigation, 2014

Background/Aims: To investigate the inhibitory effect of green tea extract, epigallocatechin gall... more Background/Aims: To investigate the inhibitory effect of green tea extract, epigallocatechin gallate (EGCG), on wild-type human leiomyoma (WT-HuLM) cells and its potential action via catechol-o-methyltransferase (COMT) activity. Methods: Cell proliferation of WT-HuLM and COMT gene-silenced HuLM (COMT-shRNA-HuLM) cells treated with 0 or 100 µM EGCG for 7 days was measured using the MTT method. Total RNA and protein were extracted from cells treated with 0 or 100 µM of EGCG for 48 h. Gene expression profiling was performed using Human Signal Transduction PathwayFinder. Proliferation cell nuclear antigen (PCNA), cyclin-dependent kinase 4 (Cdk4) and COMT protein levels were detected by Western blot analyses. COMT enzyme activity was evaluated by HPLC. Results: EGCG-treated WT-HuLM cells showed significantly decreased COMT expression (p < 0.001) and enzyme activity (p < 0.05) compared to untreated WT-HuLM cells, while COMT-shRNA-HuLM cells showed no significant change. At 100 μM of...

Molecular Carcinogenesis, 2008

Soy protein is known to have breast tumor suppressing activity. The expression of peripheral benz... more Soy protein is known to have breast tumor suppressing activity. The expression of peripheral benzodiazepine receptors (PBRs), currently renamed as translocator protein (TSPO) and their associated functions, such as nuclear cholesterol uptake and content also have been shown to be increased in breast cancer. Here we investigated whether the breast tumor suppressing effects of soy protein is mediated by down-regulation of PBR expression and function. Breast tumors were induced by gavage administration of a single dose (80 mg/kg) of dimethylbenz[a]anthracene (DMBA) into 50-d old female Sprague Dawley rats, maintained on a standard AIN-76A diet containing either casein or soy protein. Approximately 120 d following DMBA administration, the animals were sacrificed. All tumors were detected by palpation and at autopsy biopsy specimens were taken for histological grading. The ligand binding capacity, expression, and protein levels of PBRs, their nuclear localization and function, such as nuclear cholesterol uptake and content, were significantly increased in the tumors. However, replacement of casein by soy protein in the diet caused a significant decrease in all of these parameters. These data suggest that soy protein inhibits breast tumor development by decreasing the expression of the tumor-promoting gene, which encodes PBRs.

Journal of Biochemical and Molecular Toxicology, 2009

2-Choloroethyl Ethyl Sulfide (CEES) exposure causes inflammatory lung diseases, including acute r... more 2-Choloroethyl Ethyl Sulfide (CEES) exposure causes inflammatory lung diseases, including acute respiratory distress syndrome (ARDS) and pulmonary fibrosis. This may be associated with oxidative stress, which has been implicated in the desensitization of beta-adrenergic receptors (β-ARs). The objective of this study was to investigate whether lung injury induced by intratracheal CEES exposure (2 mg/kg body weight) causes desensitization of β-ARs. The animals were sacrificed after 7 days and lungs were removed. Lung injury was established by measuring the leakage of iodinated-bovine serum albumin ([ 125 I]-BSA) into lung tissue. Receptor-binding characteristics were determined by measuring the binding of [ 3 H] dihydroalprenolol ([ 3 H] DHA) (0.5-24 nM) to membrane fraction in the presence and absence of DL-propranolol (10 μM). Both high-and low-affinity β-ARs were identified in the lung. Binding capacity was significantly higher in low-affinity site in both control and experimental groups. Although CEES exposure did not change K D and B max at the high-affinity site, it significantly decreased both K D and B max at low affinity sites. A 20% decrease in β 2-AR mRNA level and a 60% decrease in membrane protein levels were observed in the experimental group. Furthermore, there was significantly less stimulation of adenylate cyclase activity by both cholera toxin and isoproterenol in the experimental group in comparison to the control group. Treatment of lungs with 3-isobutyl-1-methylxanthine (IBMX), an inhibitor of phosphodiesterase (PDE) could not abolish the difference between the control group and the experimental group on the stimulation of the adenylate cyclase activity. Thus, our study indicates that CEES-induced lung injury is associated with desensitization of β 2-AR.

Journal of Biochemical and Molecular Toxicology, 2005

Exposure to mustard gas causes inflammatory lung diseases, including acute respiratory distress s... more Exposure to mustard gas causes inflammatory lung diseases, including acute respiratory distress syndrome (ARDS). A defect in the lung surfactant system has been implicated as a cause of ARDS. A major component of lung surfactant is dipalmitoyl phosphatidylcholine (DPPC) and the major pathway for its synthesis is the cytidine diphosphocholine (CDP-choline) pathway. It is not known whether the ARDS induced by mustard gas is mediated by its direct effects on some of the enzymes in the CDPcholine pathway. In the present study, we investigated whether mustard gas exposure modulates the activity of cholinephosphotransferase (CPT), the terminal enzyme by CDP-choline pathway. Adult guinea pigs were intratracheally infused with single doses of 2chloroethyl ethyl sulfide (CEES) (0.5 mg/kg b.wt. in ethanol). Control animals were injected with vehicles only. The animals were sacrificed at different time and the lungs were removed after perfusion with physiological saline. CPT activity increased steadily up to 4 h and then decreased at 6 h and stabilized at 7 days in both mitochondria and microsomes. To determine the dose-dependent effect of CEES on CPT activity, we varied the doses of CEES (0.5-6.0 mg/kg b.wt.) and sacrificed the animals at 1 h and 4 h. CPT activity showed a dose-dependent increase of up to 2.0 mg/kg b.wt. of CEES in both mitochondria and microsomes, and then decreased at 4.0 mg/kg b.wt. For further studies, we used a fixed single dose of CEES (2.0 mg/kg b.wt.) and fixed exposure time (7 days). Lung injury was determined by measuring the leakage of iodinated-bovine serum albumin into lung tissue and expressed as the permeability index. CEES exposure (2.0 mg/kg b.wt. for 7 days) caused a significant decrease

Journal of Biochemical and Molecular Toxicology, 2006

Mustard gas exposure causes inflammatory lung diseases. Many inflammatory lung diseases are assoc... more Mustard gas exposure causes inflammatory lung diseases. Many inflammatory lung diseases are associated with oxidative stress. Reactive oxygen species (ROS) are involved in the maintenance of physiological functions. In tissues, it is therefore essential to maintain a steady-state level of antioxidant activity to allow both for the physiological functions of ROS to proceed and at the same time preventing tissue damage. We have recently reported that mustard gas exposure decreases the overall activity of superoxide dismutase (SOD). In the present study, we investigated the effects of mustard gas on each of the three isozymes: SOD-1 (Cu/Zn), SOD-2 (Mn), and SOD-3 (extracellular). Adult guinea pigs were intratracheally injected single doses of 2-chloroethyl ethyl sulfide (CEES) (2 mg/kg body weight) in ethanol. Control animals were injected with vehicle in the same way. The animals were sacrificed after 7 days, and lungs were removed after perfusion with physiological saline. Lung injury was established by measuring the leakage of iodinated-BSA into lung tissue. Mustard gas exposure caused a significant increase in the activity of SOD-1 (35%). However, the SOD-3 activity which is the predominant type in lung was significantly decreased (62%), whereas no change was observed in SOD-2 activity. Thus the decrease in the total activity of SOD was primarily due to the SOD-3 isozyme. Northern blot analysis indicated 3.5-fold increased expression of SOD-1 in mustard gas exposed lung, but no significant change in the expression of SOD-2 and SOD-3 was observed. Mustard gas exposure did not cause mutation in the coding region of SOD-1 gene while causing modulation in expression levels. The protein levels of SOD-1, SOD-2, and SOD-3 were not altered significantly in the mustard gas exposed lung. Our results indicate that

Investigative Opthalmology & Visual Science, 2006

To characterize the angiostatic effect of penetrating ocular injury and to begin to explore its m... more To characterize the angiostatic effect of penetrating ocular injury and to begin to explore its mechanism, with an emphasis on the role of pigment epithelium-derived factor (PEDF). METHODS. Using the rat model of oxygen-induced retinopathy (OIR), single or multiple dry needle injuries were made, penetrating the globe of one eye; the opposite eye served as a control. Eyes were harvested from rats killed 1, 3, and 6 days after injury, and retinas were dissected and processed for assessment of neovascularization and microglial activation or were processed for genetic and proteomic analysis. Temporal and spatial expression patterns of PEDF were analyzed by in situ hybridization. RESULTS. Penetrating ocular injury resulted in a 30% decrease in neovascular area in the retinas of OIR rats. At day 1 after injury, needle insertion caused a 4.1-fold increase in retinal PEDF mRNA and a 1.5-fold increase in retinal PEDF protein. Vitreous PEDF protein increased 3.4-fold in injured eyes compared with noninjured eyes. In situ hybridization showed an increase in PEDF mRNA in areas surrounding the puncture site. Concentrated vitreous protein from injured eyes caused a 60% decrease in retinal neovascularization when injected into the vitreous cavity of OIR rats. Preincubation of vitreous samples with anti-PEDF partially abolished this efficacy. CONCLUSIONS. The pattern of angiostasis resulting from penetrating ocular injury is consistent with the release of an endogenous antiangiogenic factor from the wound site. Preliminary studies show a possible role for PEDF in this effect. Further characterization of this role and the identification of other factors may lead to new therapeutic strategies for angiogenic eye conditions.

Investigative Opthalmology & Visual Science, 2003

PURPOSE Restructuring of extracellular matrix at actively extending blood vessel tips involves se... more PURPOSE Restructuring of extracellular matrix at actively extending blood vessel tips involves secretion of plasminogen activator (PA). Findings in earlier studies conducted in the authors' laboratory have suggested that angiostatic steroids suppress the PA activity essential for the invasive aspect of angiogenesis by increasing synthesis of plasminogen activator inhibitor (PAI)-1. This experiment was designed to test the effect of administration of exogenous PAI-1 on retinal neovascularization (NV) in an animal model of retinopathy of prematurity (ROP). METHODS At birth, Sprague-Dawley rats were placed into incubators and exposed to an atmosphere alternating between 50% and 10% O 2 every 24 hours. After 14 days, the animals were removed to room air, at which time each received a single intravitreal injection of 5 L of buffer vehicle or one of five doses of PAI-1, ranging from 3.0 g/mL to 2.0 mg/mL. Animals were killed 6 days later, and retinal NV was assessed using adenosine diphosphatase (ADPase) histochemical staining. RESULTS Retinal neovascularization decreased with increasing PAI-1 dosage. The most effective dose tested (2.0 mg/mL) caused a 52% reduction in retinal NV relative to vehicle (P Ͻ 0.005). Normal vasculogenesis, as determined by measuring retinal vascular area, was unaffected. CONCLUSIONS PAI-1 inhibits pathologic angiogenesis without adversely affecting normal vasculogenesis, an attractive feature for ROP therapies. Moreover, PAI's relationship to matrix metalloproteinases, which are also implicated in angiogenesis, suggests that the proteolytic aspect of the process may provide additional downstream therapeutic targets. (Invest Ophthal

International Journal of Women's Health, 2013

Background: Uterine fibroids (UFs, also known as leiomyoma) affect 70% of reproductive-age women.... more Background: Uterine fibroids (UFs, also known as leiomyoma) affect 70% of reproductive-age women. Imposing a major burden on health-related quality-of-life (HRQL) of premenopausal women, UF is a public health concern. There are no effective medicinal treatment options currently available for women with symptomatic UF. Objectives: To evaluate the efficacy and safety of green tea extract (epigallocatechin gallate [EGCG]) on UF burden and quality of life in women with symptomatic UF, in a double-blinded, placebo-controlled randomized clinical trial. Methods: A total of 39 reproductive-age women (age 18-50 years, day 3 serum folliclestimulating hormone ,10 mIU/mL) with symptomatic UF were recruited for this study. All subjects had at least one fibroid lesion 2 cm 3 or larger, as confirmed by transvaginal ultrasonography. The subjects were randomized to oral daily treatment with either 800 mg of green tea extract (45% EGCG) or placebo (800 mg of brown rice) for 4 months, and UF volumes were measured at the end, also by transvaginal ultrasonography. The fibroid-specific symptom severity and HRQL of these UF patients were scored at each monthly visit, using the symptom severity and quality-of-life questionnaires. Student's t-test was used to evaluate statistical significance of treatment effect between the two groups. Results: Of the final 39 women recruited for the study, 33 were compliant and completed all five visits of the study. In the placebo group (n = 11), fibroid volume increased (24.3%) over the study period; however, patients randomized to green tea extract (n = 22, 800 mg/day) treatment showed significant reduction (32.6%, P = 0.0001) in total UF volume. In addition, EGCG treatment significantly reduced fibroid-specific symptom severity (32.4%, P = 0.0001) and induced significant improvement in HRQL (18.53%, P = 0.01) compared to the placebo group. Anemia also significantly improved by 0.7 g/dL (P = 0.02) in the EGCG treatment group, while average blood loss significantly decreased from 71 mL/month to 45 mL/month (P = 0.001). No adverse effects, endometrial hyperplasia, or other endometrial pathology were observed in either group. Conclusion: EGCG shows promise as a safe and effective therapeutic agent for women with symptomatic UFs. Such a simple, inexpensive, and orally administered therapy can improve women's health globally.

Injury Extra, 2007

, and whose medical notes were available were included in the audit. Results: Sixty-eight patient... more , and whose medical notes were available were included in the audit. Results: Sixty-eight patients were available for analysis. They included 44 (65%) male and 24 (35%) female with mean age of 36 (2-101). There were varying grades of injury: 24% Gustillo and Anderson Grade I, 39% Grade II, 16% Grade IIIa, 18% IIIb and 3% IIIc. Only 3% of cases had documented evidence of appropriate wound care in A&E. Ninety-three percent of patients received IV antibiotics given at a mean time from injury of 2 h 37 min. Of the 44% of cases referred to plastics, only 13% (4/30) of referrals were made before the initial procedure. The mean time from injury to initial operative procedure was 9 h 34 min (median 5 h 22 min). Only 23% of cases were carried out with a consultant present. Ten of 62 cases (six lost to follow-up) were complicated by infection, 2 (3%) of which were deep. Two cases of compartment syndrome occurred requiring fasciotomy. All fractures united with a mean time to union of 39 weeks. The most striking problem was the poor quality of note keeping and hence difficulty in obtaining accurate data. Conclusions: The results show that in a number of crucial areas the guidelines are not adhered to. However, the complication rate in terms of nonunion and deep infection is respectable compared to current literature. One cannot read too much into this due to the problems with data collection. The department should examine its current protocol and perform a prospective audit thereby obtaining an accurate picture of its results. In the meantime emergent debridement of soft tissues, fracture stabilisation and early administration of intravenous antibiotics should be supported.

Human Reproduction, 2013

Is targeted adenovirus vector, Ad-SSTR-RGD-TK (Adenovirus-human somatostatin receptor subtype 2-a... more Is targeted adenovirus vector, Ad-SSTR-RGD-TK (Adenovirus-human somatostatin receptor subtype 2-arginine, glycine and aspartate-thymidine kinase), given in combination with ganciclovir (GCV) against immortalized human leiomyoma cells (HuLM) a potential therapy for uterine fibroids? summary answer: Ad-SSTR-RGD-TK/GCV, a targeted adenovirus, effectively reduces cell growth in HuLM cells and to a significantly greater extent than in human uterine smooth muscle cells (UtSM). what is known already: Uterine fibroids (leiomyomas), a major cause of morbidity and the most common indication for hysterectomy in premenopausal women, are well-defined tumors, making gene therapy a suitable and potentially effective non-surgical approach for treatment. Transduction of uterine fibroid cells with adenoviral vectors such as Ad-TK/GCV (herpes simplex virus thymidine kinase gene) decreases cell proliferation. study design, size, duration: An in vitro cell culture method was set up to compare and test the efficacy of a modified adenovirus vector with different multiplicities of infection in two human immortalized cell lines for 5 days. participants/materials, setting, methods: Immortalized human leiomyoma cells and human uterine smooth muscle cells were infected with different multiplicities of infection (MOI) (5-100 plaque-forming units (pfu)/cell) of a modified Ad-SSTR-RGD-TK vector and subsequently treated with GCV. For comparison, HuLM and UtSM cells were transfected with Ad-TK/GCV and Ad-LacZ/GCV. Cell proliferation was measured using the CyQuant assay in both cell types. Additionally, western blotting was used to assess the expression of proteins responsible for regulating proliferation and apoptosis in the cells. main results and the role of chance: Transduction of HuLM cells with Ad-SSTR-RGD-TK/GCV at 5, 10, 50 and 100 pfu/cell decreased cell proliferation by 28, 33, 45, and 84%, respectively (P , 0.05) compared with untransfected cells, whereas cell proliferation in UtSM cells transfected with the same four MOIs of Ad-SSTR-RGD-TK/GCV compared with that of untransfected cells was decreased only by 8, 23, 25, and 28%, respectively (P , 0.01). Western blot analysis showed that, in comparison with the untargeted vector Ad-TK, Ad-SSTR-RGD-TK/GCV more effectively reduced expression of proteins that regulate the cell cycle (Cyclin D1) and proliferation (PCNA, Proliferating Cell Nuclear Antigen), and it induced expression of the apoptotic protein BAX, in HuLM cells. limitations, reasons for caution: Results from this study need to be replicated in an appropriate animal model before testing this adenoviral vector in a human trial. wider implications of the findings: Effective targeting of gene therapy to leiomyoma cells enhances its potential as a noninvasive treatment of uterine fibroids.

Gynecologic and Obstetric Investigation, 2013

Postoperative abdominal/pelvic peritoneal adhesions are a major source of morbidity (bowel obstru... more Postoperative abdominal/pelvic peritoneal adhesions are a major source of morbidity (bowel obstruction, infertility, ectopic gestation as well as chronic pelvic pain) in women. In this study, we screened various transduction and transcription modifications of adenovirus (Ad) to identify those that support maximal Ad-mediated gene delivery to human adhesion fibroblasts, which in turn would enhance the efficacy of this novel treatment/preventative strategy for postoperative adhesions. We transduced primary cultures of human peritoneal adhesion fibroblasts with fiber-modified Ad vectors Ad5-RGD-luc, Ad5-Sigma-luc, Ad5/3-luc and Ad5-CAV2-luc as well as transcriptional targeting viruses Ad5-survivin-luc, Ad5-heparanase-luc, Ad5-mesothelin (MSLN)-CRAd-luc and Ad5-secretory leukoprotease inhibitor (SLPI)-luc, and compared their activity to wild-type Ad5-luc. At 48 h, luciferase activity was measured and normalized to the total protein content in the cells. Among the fiber-modified Ad vecto...

Fertility and Sterility, 2010

OBJECTIVE: Mullerian and male urogenital anomalies share common genetic, hormonal and environment... more OBJECTIVE: Mullerian and male urogenital anomalies share common genetic, hormonal and environmental etiologic factors. The purpose of this study is to quantify the risk of urogenital anomalies in male relatives of women with Mullerian anomalies. DESIGN: Kinship analysis. MATERIALS AND METHODS: Patients were identified by ICD codes from the largest hospitals in the state of Utah (1994 to 2009). We included all patients who were coded for doubling of the uterus or Mullerian anomalies. Controls for the probands were matched based on birth year and sex (1:5). All records were subsequently matched to the Utah population data base (UPDB) to obtain pedigrees with inclusion of data regarding male urogenital anomalies and to perform kinship analysis. The prevalence of the following urogenital anomalies (undescended testis, hypospadias, epispadias, anomalies of the penis, or other male genital anomalies) in all male relatives of the probands was compared to controls. The Kinship analysis software kingless was used to compute the relative risks for urogenital anomalies among male relatives of probands with female Mullerian anomalies. RESULTS: The probands comprised 2939 women with Mullerian anomalies. Males with urogenital anomalies identified through ICD codes were 10176 patients. First degree (children, parent) male relatives of women with Mullerian anomalies had a RR of urogenital anomalies of 1.43 (1.09-1.89). This risk was largely limited to children of the probands (RR¼1.47, 1.1-1.97). No parents of the probands or controls were affected. Relative risks for the other kinship classes were not significantly elevated. The prevalence of anomalies in male children of women with Mullerian anomalies were as follows: undescended testis (53%),hypospadias (36%), epispadias (0%), anomalies of the penis (29%) and other male genital anomalies (1.5%). CONCLUSION: Urogenital anomalies appear to be increased in male children of women with Mullerian anomalies. Undescended testis and hypospadias are the most common anomalies. Supported by: Departmental. OBJECTIVE: Uterine fibroids are the most common benign tumors of women. Fibroids are more common in African American (AA), and the prevalence increases with age, but little is known about other races. We examined the effects of age, race, BMI, and hormonal markers on fibroid prevalence in a large, multi-ethnic population. DESIGN: Cross-sectional.

Experimental Eye Research, 2004

Retinal capillary quiescence is regulated by a delicate balance between proangiogenic and anti-an... more Retinal capillary quiescence is regulated by a delicate balance between proangiogenic and anti-angiogenic factors. Pathological angiogenesis is the result of a shift in this balance towards proangiogenic influences. Pathological angiogenesis is produced in a rat model of oxygen-induced retinopathy (OIR) by exposing newborn rat pups to alternating periods of hyperoxia and hypoxia. Based upon previous work, two similar exposure paradigms were investigated and compared, exposure of rat pups to alternating periods of 45 and 12$5% oxygen, and to alternating periods of 40 and 15% oxygen. The resulting retinal pathology was assessed by measurement of retinal clock hours with pathological blood vessel growth and the percentage of the retina that is avascular. The 45 and 12$5% exposure produced significantly greater incidence and severity of pathology than the 40 and 15% protocol. To explain the difference in pathology between these two very similar exposure protocols, retinal levels of proangiogenic vascular endothelial growth factor (VEGF) and VEGF receptor 2 (VEGFR2) and anti-angiogenic pigment epithelium-derived factor (PEDF) were measured by ELISA and western blot analysis at 0, 2, and 6 days postexposure. In whole retinal lysates, there were no significant differences in VEGFR2 and PEDF levels. However, VEGF levels were approximately 48 and 78% higher on post-oxygen exposure day 0 and 2, respectively, in the group treated with alternating periods of 45 and 12$5% oxygen compared to the group treated with alternating periods of 40 and 15% oxygen. There was no significant difference in VEGF levels between these two groups on day 6 post-exposure. Therefore, the difference in pathology observed between these two experimental paradigms is associated with differences in whole retinal VEGF levels, but not changes in whole retinal VEGFR2 or PEDF levels. The results of this study suggest the existence of a threshold in the rat model of OIR, such that a small change in blood oxygen profile triggers a disproportionate increase in subsequent neovascularization, which is accompanied by more dramatic changes of retinal VEGF level than VEGFR2 or PEDF level. If a similar threshold exists for humans, it could explain why some oxygen-treated premature infants develop retinopathy and others do not, despite similar gestational ages, birth weights and clinical courses.

American Journal of Obstetrics and Gynecology, 2010

Objective-Investigate the effect of epigallocatechin gallate (EGCG), on rat leiomyoma (ELT3) cell... more Objective-Investigate the effect of epigallocatechin gallate (EGCG), on rat leiomyoma (ELT3) cells in vitro and in nude mice model. Study Design-ELT3 cells were treated with various concentrations of EGCG. Cell proliferation, PCNA and Cdk4 protein levels were evaluated. ELT3 cells were inoculated subcutaneously in female athymic nude mice. Animals were fed 1.25mg EGCG (in drinking water)/mouse/day. Tumors were collected and evaluated at 4 and 8 weeks post-treatment. Results-Inhibitory effect of EGCG (200 μM) on ELT3 cells was observed after 24 h treatment (p<0.05). At ≥50μM, EGCG significantly decreased PCNA and Cdk4 protein levels (p<0.05). In vivo, EGCG treatment dramatically reduced the volume and weight of tumors at 4 and 8 weeks posttreatment (p<0.05). The PCNA and Cdk4 protein levels were significantly reduced in EGCG treated group (p<0.05). Conclusion-EGCG effectively inhibits the proliferation and induce apoptosis in rat ELT3 uterine leiomyoma cells in vitro and in vivo.

Experimental Eye Research, 2004

The pathogenesis of retinopathy of prematurity involves dysregulated angiogenesis resulting in pr... more The pathogenesis of retinopathy of prematurity involves dysregulated angiogenesis resulting in pre-retinal growth of new vessels. Inhibition of tyrosine kinase-dependent pro-angiogenic signals may provide a rational therapeutic approach to the reduction of pre-retinal neovascularization. Vascular endothelial growth factor stimulates endothelial cell mitogenesis, differentiation and migration, by binding and activating the receptor tyrosine kinases vascular endothelial growth factor receptor-1 and vascular endothelial growth factor receptor-2. One of the vascular endothelial growth factor receptor substrates implicated in vascular endothelial growth factor signal transduction is c-Src. The ability of herbimycin A, a c-Src-selective tyrosine kinase inhibitor, to inhibit vascular endothelial growth factor-induced bovine retinal microvascular endothelial cell proliferation and tube formation was investigated. The ability of the compound to inhibit pathologic angiogenesis was tested in a rat model of retinopathy of prematurity. Exposure of neonatal rats to oxygen concentrations cycling between 10 and 50% induced severe pre-retinal neovascularization in all rats. Some of the eyes of these variable oxygen-exposed rats were herbimycin A-injected or vehicle-injected 1 or 3 days post-oxygen exposure while some eyes were non-injected. All rats were sacrificed for assessment 6 days post-exposure. Herbimycin A inhibited both vascular endothelial growth factor-induced bovine retinal microvascular endothelial cell proliferation and capillary tube formation in a dose-dependent manner. Injection of herbimycin A into oxygen-treated rats 1 day post-oxygen exposure produced a 63% decrease in pre-retinal neovascularization relative to vehicle (P = 0.0029). There was a 41% decrease in pre-retinal neovascularization in herbimycin-injected eyes relative to vehicle-injected eyes 3 days post-oxygen (P = 0.031). Pre-retinal neovascularization was reduced in vehicle-injected eyes relative to non-injected eyes at both injection times. There were no significant differences in retinal vascular area between any of the experimental groups. Based on the results of this study, herbimycin A inhibits endothelial cell proliferation and tube formation at non-toxic concentrations and reduces pre-retinal neovascularization in a rat model of retinopathy of prematurity. Reduction of angiogenesis by the inhibition of tyrosine kinase activity may be a viable route to the development of effective chemotherapies applicable to eye disease.