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Once bullied, Cleveland Browns rookie Garrett Gilkey wants to help kids with similar experiences
By Tom Reed, Cleveland.com
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Garrett Gilkey grew into a small-college powerhouse for Chadron State, earning a seventh-round draft pick from the Browns. But he says he was a victim of bullying as a teenager, and is eager to help other youngsters dealing with that problem.
(Dave Martin, Associated Press)
BEREA, Ohio -- When Garrett Gilkey places his hand in the dirt and gazes across the line of scrimmage, he doesn't picture the faces of the tormentors who reduced him to tears during his freshman year in high school.
The Browns' seventh-round draft pick doesn't conjure images of tumult -- the bullying, the taunts, the pranks he says he endured -- to motivate him at the snap of the ball. The offensive lineman concedes, however, the experiences in the school hallways and streets of a rural Illinois town nine years ago benefit him in one capacity when he buckles his chinstrap.
"The person across the line wants to put me on my back and I'm not going to let him do it," Gilkey said in a post-draft phone interview. "I don't bring back those old feelings from my freshman year when I'm in practice or in a game. It's more of an overall mentality: I'm not going to let someone get the better of me; I'm not going to be beaten down and defeated and humiliated the way I was back then."
Some anti-bullying experts say the shame lingers long after the harassment ends. The skinny freckle-faced underclassmen who grew into a 6-6, 318-pound NFL prospect is becoming an advocate for students who fear what the next school day might bring.
Gilkey does not repress his memories. He shares them, sometimes in startling detail. He speaks of being booed during a 2005 school assembly after his name was announced, a reaction that sent him running to the restroom, his eyes welling with tears.
At the NFL Scouting Combine in late February, the converted tackle from Division II Chadron State in Nebraska sat patiently and answered questions about the most traumatic time in his life. In recent weeks, the hulking man with the flowing red mane has addressed church groups and schools. He hopes to do the same in Cleveland.
"It's about building relationships with people and tearing down barriers," he said. "Whether it's dealing with bullies or drug problems or problems in the home, it's about having perseverance. Things will happen in our lives that will be really hard. What's important is having faith and trying to persevere through them."
Anti-bullying campaigns have gained momentum in the post-Columbine era. Paulie Velotta, a Mentor-based trainer for a national prevention program, said one in every six students report being bullied. She also cited a 2010 statistic that roughly 160,000 kids miss school each day over such concerns.
She said schools need to develop comprehensive programs, and that speakers like Gilkey offer an excellent supplement.
"Absolutely, kids look up to figures such as pro athletes, entertainers, politicians, anyone in the limelight," said Velotta, a national trainer for the Olweus Bullying Prevention Program who also works at Crossroads, a behavioral health agency. "They want someone who is a positive role model. He's someone who can connect with kids because he shares their back story."
A late-round pick coming from a small school faces long odds of making an NFL roster. But Gilkey's shell has been hardened by adversity and detractors. He can't wait for the start of Browns rookie minicamp on Friday.
Troubled times
Garrett Gilkey impressed scouts at the Senior Bowl, showing the ability to battle players from much larger programs. "He's certainly has the size and good feet," says Senior Bowl executive director Phil Savage, a former Browns GM. "Can he adjust to playing (guard) at that level? The potential is there."
Escaping the suburban sprawl of Chicago, Catherine and Cary Gilkey moved their four children a decade ago to Sandwich, a small town about 60 miles southwest of Chicago, where they bought a large home with swimming pool.
Their only son played middle school football for two years and seemed to be adjusting to new environs. But in the summer prior to his freshman year, Gilkey experienced a rapid heart beat, diagnosed as Wolff-Parkinson-White syndrome. He underwent surgery and was instructed to take it easy through the fall.
Doctors orders came with unintended consequences, however. Players from the high school football team began to ostracize Gilkey, he said, because he hadn't sacrificed alongside them during the grueling practices. He filled his free time with other extra-curricular activities, but soon found himself targeted in the halls and classrooms.
Catherine told her 5-7, 170-pound son to use his intellect and big vocabulary rather than his fists to resolve disputes. But a freshman talking back to upperclassmen did not endear him to his schoolmates. Unable to participate in gym class due to his heart condition, Gilkey worked as an office aide and became the first person a student encountered on his way to a meeting with a principal.
"I became the face of discipline," Gilkey said. "You make enemies that way."
Hilary Gilkey, 25, a junior during the 2004-05 school year, said she witnessed some of the harassment her brother endured. Sometimes, she intervened, telling the agitators to "cut it out, that's my brother." Mostly, she felt helpless.
"He came home from school hurt and that pain affected all of us," Hilary said. "He became introverted and tried to brush it off."
Gilkey said he never harbored thoughts of suicide or exacting violent revenge, but each day brought challenges. He recalls being punched and shoved into lockers. A fellow student urinated in his baseball glove. While his parents alerted administrators, some of the actions took place off school grounds.
Walking home from school one day a group of kids summoned Gilkey and asked to examine his dental work – a brace had been inserted into the roof of his mouth to expand its width. As he tilted his head, one boy inserted a condom, Gilkey said.
"There was an unbelievable amount of hatred that people had for him at the school," former classmate Matt Cervantes told ESPN.com. "Sometimes, he would talk trash, and they would get on him so bad."
Cervantes declined an interview with The Plain Dealer due to the backlash over his ESPN remarks. The superintendent for the Sandwich school district did not return a call from the newspaper. Neither did a middle school counselor who Gilkey said has supported him through the years.
In the ESPN article, which generated more than a 1,000 online responses, several anonymous commentors who said they attended Sandwich at the same time as Gilkey bashed him and claimed the story was one-sided.
"Really, I like to tell people that I was just the little redheaded, gingery, skinny-looking (kid)," Gilkey told reporters at the combine. "I was pear-shaped. I had these wide hips and this skinny-looking upper body. I was just a prime target for many of the cruel kids in Sandwich."
The low points of the 2004-05 school year for Hilary were hearing her brother booed at two school assemblies.
"That's something I will never forget," said Hilary, now a teacher's assistant at a school in Aurora, Ill. "It was horrible and it was hard seeing people you knew, people that were in your class, be among those who booed. It was a very strange situation. I don't remember any staff members intervening or trying to step up."
Garrett Gilkey with his pastor, Charlie Grenade, whom he credits for helping him deal with the bullying issues of his youth and the temptation of steroids as a potential NFL prospect. "We tried to encourage him not to let his value get wrapped up (just in football)," Grenade said.
Change of scenery, fortune
As Hilary completed her final year at Sandwich, her brother enrolled at Aurora Christian, a private school more than 20 miles from home. Gilkey immediately began to make friends and played for the football team coached by former NFL receiver Don Beebe. Although his social life improved, Gilkey considered quitting the team during his sophomore year.
"He was not very coordinated at first," said Dave Beebe, an assistant coach and brother of Don. "His body was growing faster than his joints. ... You could tell he had a troubled past. We could see there was some baggage and scars, and that was OK.
"We wanted him to feel accepted and know it was OK to make mistakes. You try to get kids to see things they do not see in themselves."
Gilkey's confidence sprouted as his body developed. He shot up to 6-6, 250 pounds by his senior season, shedding his pear-shape husk.
With no Division I college offers, he took the advice of Don Beebe, who pointed him to his alma mater, Chadron State. He thrived under the tutelage of coach Bill O'Boyle and local pastor Charlie Grenade. He spent his final two years in Nebraska living with the pastor and his wife, studying scripture and learning how to resist temptation.
Gilkey credits Grenade for helping him steer clear of steroids, an option he considered as an NFL career became a possibility, and for maintaining a positive outlook after an appendectomy cost him two games his junior season.
"He thought (the appendectomy) was his second strike against him," Grenade said. "We tried to encourage him not to let his value get wrapped up (just in football)."
NFL scouts found their way to remote the campus in northwest Nebraska and watched as the tackle dominated his low-level competition. He played well enough to earn invitations to the Senior Bowl and combine.
Senior Bowl executive director Phil Savage likes Gilkey, but sees him as a developmental player.
"He's certainly has the size and good feet," said Savage, a former Browns general manager. "He's a bit of a waist bender and he's got to learn to get his head and weight back. The question is can he adjust to playing (guard) at that level? The potential is there."
Giving voice to those lacking it
The Gilkeys and friends gathered last Saturday at the family home in Sandwich, a place wrought with anxiety nine years earlier. A different kind of tension filled the house last week, a joyous suspense as the draft's later rounds unfurled. When the Browns called and he became the 227th pick, Gilkey played a joke on his guests.
With cellphone to ear, he pantomimed to friends and family the call wasn't about the draft. Then, he hung up and told everyone he was a Cleveland Brown.
The room erupted. Hilary said the prank was the type of goofiness she remembered from her brother before his freshman year in high school.
Gilkey said he still sees some of his tormentors around town. He bears no grudges, he says, and refuses to divulge names.
While Ohio has enacted anti-bullying legislation for schools, Gilkey's home state fell several votes shy a year ago.
"It's a very real thing that people experience," he said. "There are those who still believe that this happens to only 'soft kids' and it all can be fixed by 'toughening them up.'"
Baltimore Ravens halfback Ray Rice is one of several current and former NFL players to advocate on behalf of bullied children. Gilkey hopes to join the ranks. He wants to engage bystanders, the ones who witness bullying, "the people who can give a voice to those who don't have one." Students who see it need to either intervene, he said, or alert school officials.
Gilkey plans to become a youth minister after his playing days end. When he speaks to groups, his size always becomes a talking point.
"People want to know 'How did this giant man get bullied?'" he said. "I think it makes the story more beautiful, and it helps eliminates some of the stigmas. It's OK for big, tough linemen to share their experiences. We're people. We all have emotions."
Garrett Gilkey is not afraid to show his vulnerable side. Right up to the moment he buckles the chinstrap.


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Biol. Res. vol.52 Santiago 2019 Epub 28-Nov-2019
Gramicidin inhibits human gastric cancer cell proliferation, cell cycle and induced apoptosis
Tingting Chen1 †
Yong Wang2 †
Yang Yang1
Kaikai Yu3
Xiangliao Cao1
Fang Su1
Huanbai Xu4
Yongde Peng4
Yudong Hu3 *
Feng Qian1 3 *
Zishu Wang1 *
1Department of Medical Oncology, First Affiliated Hospital of Bengbu Medical College, Anhui Province Key Laboratory of Translational Cancer Research (Bengbu Medical College), 287 Changhuai Road, Bengbu 233004, Anhui Province, People's Republic of China
2Department of General Surgery, Zhoupu Hospital affiliated to Shanghai Health Medical College, Shanghai 201318, People's Republic of China
3Engineering Research Center of Cell & Therapeutic Antibody, Ministry of Education, School of Pharmacy, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai 200240, People's Republic of China
4Department of Endocrinology and Metabolism, Shanghai General Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200080, People's Republic of China
Gastric cancer is a common malignant tumor with high morbidity and mortality worldwide, which seriously affects human health. Gramicidin is a short peptide antibiotic which could be used for treating infection induced by bacteria or fungi. However, the anti-cancer effect of gramicidin on gastric cancer cells and its underlying mechanism remains largely unknown.
Gastric cancer cells SGC-7901, BGC-823 and normal gastric mucosal cells GES-1 were treated with different concentrations of gramicidin respectively. The results of CCK-8 experiment revealed cellular toxicity of gramicidin to cancer cells while cell colony formation assay showed that gramicidin significantly inhibited the proliferation of gastric cancer cells, but had little effect on normal gastric mucosal cells. In addition, the wound healing assay showed that gramicidin inhibited the migration of SGC-7901 cell. Meanwhile, apoptosis and cell cycle analysis revealed that gramicidin induced cell apoptosis with G2/M cell cycle inhibition. Furthermore, western blot analysis demonstrated that gramicidin down-regulated the expression of cyclinD1 and Bcl-2 as well as the FoxO1 phosphorylation.
The current study illustrated the anti-tumor activity of gramicidin on gastric cancer cells, providing a possibility for gramicidin to be applied in clinical practice for the treatment of gastric cancer.
Keywords: Gramicidin; Proliferation; Apoptosis; Cell cycle arrest; SGC-7901
Gastric cancer (GC) is an important health problem worldwide and is the third leading cause of cancer-associated mortality [1]. Although the incidence rate of GC is decreasing, particularly in Asian countries [2], it still causes a large proportion of GC-related death in 5 years with rapid tumor development and poor prognosis. Currently, the clinical treatment of GC is comprehensively based on surgical resection and chemotherapy [3]. Although advanced chemotherapy may improve the survival rate, they have strong side effects [4]. Thus, finding effective drugs which have significant anti-gastric carcinoma effect are of great need undoubtedly.
Forkhead box protein O transcription factor family (FoxO) consists of four members, including FoxO1, FoxO3a, FoxO4 and FoxO6 [5] and plays a pivotal role in diverse biological functions such as cell cycle progression, cell apoptosis, glucose metabolism, and cell differentiation [6]. FoxO1 is widely expressed in tissues and regulates cyclin D1 and p21 [7] while elevated levels of phosphorylation of FoxO1 is associated with tumorigenesis [8]. Thus, FoxO1 is expected to be a potential target for anti-cancer drugs.
Gramicidin, a short peptide antibiotic derived from bacteria called Bacillus brevis, is composed of 15 l- and D-amino acid residues [9]. It is one of the very first antibiotics to be used in clinical practice [10] and has the most significant antibacterial effect on gram-positive bacteria, fungi and protozoa [11, 12]. Gramicidin mainly changes the permeability of the plasma membrane, and destroys the structure of the membrane causing Na+ influx/K+ efflux and depolarization of transmembrane potential [13, 14]. Gramicidin is often used as a membrane-active drug to study the structure and function of the membranes. Previous studies on renal cell carcinoma (RCC) found that gramicidin as an inhibitor of hypoxia-inducible factor (HIF), suppressed the cell growth and angiogenesis of renal cell carcinoma (RCC) which expresses the Von Hippel-Lindau (VHL) [15]. However, the function of gramicidin on gastric cancer has not been reported.
In this study, we focused on evaluating the possible anti-tumor effects of gramicidin in GC while investigating the possible underlying mechanism. Our study found that the proliferation of gastric cancer cells was decreased significantly upon gramicidin treatment with induction of apoptosis and inhibition of migration and cell cycle. Further analysis revealed that FoxO1 signaling and Bcl-2, cyclin D1 might be modulated.
Gramicidin liquid preparation
10 μg gramicidin dissolved in DMSO to prepare a solution of 100 mmol/ml and stored at – 20 °C for long-term. Then the solution was diluted to different concentrations with DMSO for experiment or stored at – 20 °C.
Cell lines and culture condition
Human gastric cancer cell lines SGC-7901 and BGC-823 were reserved by our laboratory. Human normal gastric mucosal cell lines GES-1 were bought from the cell bank of the Chinese Academy of Sciences (Shanghai, China). All the cell lines were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin. The cells were grown in a humidified 37 °C incubator with 5% CO2.
Fetal bovine serum (FBS) was purchased from Hangzhou Sijiqing Biological Engineering Materials Co., Ltd. (Hangzhou, China). RPMI-1640, penicillin/streptomycin and phosphate buffer solution (PBS) were obtained from Gibco (Grand Island, USA). Dimethyl sulfoxide (DMSO) was bought from Sigma-Aldrich (St. Louis, USA). Wright-Giemsa Stain Kit was obtained from Nan Jing Jian Cheng Technology Company (Nanjing, China). Annexin V-FITC apoptosis detection kit and Cell counting kit-8 was provided by Beyotime Institute of Biotechnology (Shanghai, China). Antibodies against p-FoxO1, total- FoxO1, cyclin D1, Bcl-2 and GAPDH were purchased from Cell Signaling Technology, Inc. (Danvers, USA). HRP-conjugated secondary antibodies against rabbit and mice were supplied from Jackson ImmunoResearch Inc. (West Grove, USA). Methanol and ethanol were bought from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China).
Colony formation assay
To observe the effect of gramicidin on proliferation of gastric cancer cells SGC-7901 and BGC-823 and normal gastric mucosa cells GES-1 cells. Cells in logarithmic growth phase were seeded in 6-well plates (500 cells/well), then cells were treated with DMSO (as a control group) and different concentrations of gramicidin (10 nM, 20 nM, 30 nM, 40 nM and 50 nM) with RPMI 1640 containing 10% FBS and incubated for 7-10 days until they reached optimal clones of 200 cells in each clone. The medium was discarded and methanol was added to each well for 20 min. Cells were stained with Giemsa for another 20 min and washed with water. The number of colonies in each well was counted. The data obtained were analyzed by Prism software (GraphPad Software, Inc.).
Cell counting kit-8 assay
SGC-7901 cells (5000 cells/well) were cultured in a 96-well plate overnight and treated with different concentrations of gramicidin (0.1 μM, 0.3 μM, 1 μM, 3 μM, 10 μM, 30 μM and 100 μM). DMSO was used as a control. After 24 h, 10 μl of CCK-8 solution were added to each well for 2 h then the 450 nm-absorbance was measured by an microplate reader.
The wound healing assay was applied to assess cell migration. Well-grown SGC-7901 cells were seeded in 12 well plates (5 × 105 cells/well) in RPMI 1640 supplemented with 5% FBS and incubated under 37 °C and 5% CO2 condition for 24 h. When the cell fusion reached 90%, the wound was scratched with a sterile 200 μl tip. Then cells were washed with PBS for three times, and the corresponding concentration of gramicidin (0 μM, 0.3 μM, 1 μM and 3 μM) were added into each well. Photographs were taken at 0 h and 48 h respectively and the changes in the healing of scratches of each well were recorded. Finally, Image J was employed to compute the area covered by the cells, and the degree of decline in the healing rate after administration was calculated as compared with the DMSO solvent control group.
Cell apoptosis assay
SGC-7901 cells in logarithmic growth phase were treated with trypsin without EDTA, and were seeded in 6-well plates with 106 per well, then were incubated at 37 °C and 5% CO2 condition overnight. Gramicidin (0 μM, 0.3 μM, 1 μM and 3 μM) were added into corresponding well for 24 or 48 h. The cells were harvested, rinsed with pre-cooling PBS twice, then stained with Annexin V-FITC and PI for 5 min according to the manufacturer's instructions. Flow cytometry was used to analyze the stained cells within 1 h. The data was analyzed by the FlowJo7.6 software.
Cell cycle assay
SGC-7901 cells (5 × 105/well) were cultured in 6-well plates and incubated with different concentrations of gramicidin (0 μΜ, 0.3 μΜ, 1 μΜ and 3 μΜ) for 24 h or 48 h. The cells were harvested and rinsed with cold PBS before cells were resuspended in 70% cold ethanol at 4 °C overnight. After centrifugation to remove ethanol, the cells were washed with pre-cold PBS for twice, treated with 10 μg/ml RNAse at 37 °C for 30 min and stained with 50 μg/ml PI solution in the dark for 5 min. Finally, samples were analyzed by flow cytometry (LSR-Fortessa TM X-20; BD Biosciences, San Jose, NJ, USA).
SGC-7901 cells (1 × 106/well) were collected after being treated with gramicidin or DMSO for 48 h. Cells were then washed twice with ice-cold PBS. Then, loading buffer was added to the wells to extract protein. Protein lysates were centrifuged slightly and heated at 99 °C for 10 min. The lysates were fractionated by electrophoresis in 12% or 15% SDS-PAGE. Then protein lysates were transferred to nitrocellulose membranes and blocked with 5% non-fat milk in water for 2 h in room temperature. The membranes were incubated with primary antibodies at 1:1000 dilutions overnight at 4 °C with continuous agitation. Then the membranes were incubated with secondary antibody for 2 h at room temperature, followed by being washed three times. The proteins were visualized on the X-ray film. The densitometry analysis of band intensity was detected by ImageJ. The experiment was repeated three times independently.
Data analysis and statistics
All values were presented as the mean ± SEM. Statistical significance of the results was performed with t test and one-way analysis of variance (ANOVA) using Graphpad Prism 5.0. Statistically significant P-values were defined as *P < 0.05 and **P < 0.01, ***P < 0.005. The chemical structure of gramicidin was presented by ChemDraw Professional 16.0 software.
Cytotoxic effect of gramicidin on the gastric cancer
The chemical structure of gramicidin was shown in the Fig. 1a. To determine whether gramicidin exert cytotoxic effect on human gastric cancer SGC-7901 and BGC-823 cells, cell counting kit-8 assay was applied and the cells were treated with different concentrations of gramicidin for 24 h. As shown in Fig. 1b, c, the percent of living cells decreased significantly upon gramicidin treatment and gramicidin inhibited the proliferation of two different kinds of gastric cancer cells in a dose-dependent manner. The 50% inhibitory concentration (IC50) values of gramicidin, were 0.183 and 0.191 μΜ for the SGC-7901 and BGC-823 cells, respectively. In addition, results showed that SGC-7901 cells was more sensitive to the treatment of gramicidin.
Fig. 1 The chemical structure of gramicidin and its toxic effect on gastric cancer cells SGC-7901 and BGC-823 cells proliferation. a Chemical structure of gramicidin. The cell survival rate of b SGC7901 and c BGC-823 cells which were treated with 0, 0.3, 1,3, 10 and 30 μM of gramicidin respectively in 96-well plate were quantitatively analyzed by CCK-8 assay. The results are shown as the mean ± SEM of three independent experiments (n = 3, *P < 0.05, **P < 0.01 and ***P < 0.001 vs. Control)
Effect of gramicidin on the cell proliferation
Cell proliferation plays important role in cancer development. We then investigated the anti-proliferative effect of gramicidin on human gastric cancer cells and colony formation assay was used. As shown in the Fig. 2a, cells were treated with gramicidin at various concentration for 10 days and the colony formation rate of SGC-7901 and BGC-823 cells decreased significantly. Quantitative analysis of the clone formation rate showed that gramicidin suppressed proliferative capacity of SGC-7901 and BGC-823 cells in a concentration-dependent manner (Fig. 2b, c). However, the proliferation of human gastric mucosal epithelial cells GES-1 was not affected by gramicidin when compared to the control group (Fig. 2d). Only when the concentration of gramicidin reached to 40 nM, the proliferation of the GES-1 cells was inhibited (P < 0.05). The above results suggested that the gramicidin could inhibit the proliferation of the gastric cancer cells SGC-7901 and BGC-823. As SGC-7901 showed a more sensitive pattern upon gramicidin treatment, we next evaluate further anti-tumor effect of gramicidin on GC using the SGC-7901 cells.
Fig. 2 Inhibitory effect of gramicidin on gastric tumor SGC-7901, BGC-823 and GES-1 cells proliferation. Representative images of colonies in a SGC-7901, BGC-823 and GES-1 cells and quantification of the colony formation rate in b SGC-7901, c BGC-823 and d GES-1 cells from a six-well plate using colony formation assay while cells were treated with 0, 10, 20, 30 and 40 nM of gramicidin for 10 days, respectively. The results are shown as the mean ± SEM of three independent experiments (n = 3, *P < 0.05, **P < 0.01 and ***P < 0.001 vs. Control)
Gramicidin induced the apoptosis of human gastric cancer cells
Furthermore, to determine whether gramicidin induced apoptosis of human gastric cancer cells, Annexin V-FITC/propidium iodide (PI) double staining was performed. SGC-7901 cells were cultured with various concentrations of the gramicidin for 24 h or 48 h and flow cytometry was used to analyze the cell apoptosis. As demonstrated in Fig. 3a, c, gramicidin promoted apoptosis of SGC-7901 cells. The apoptotic rate of the gramicidin-treated cancer cells was increased remarkably in 48 h (Fig. 3b, d). When the cells were cultured with gramicidin for 48 h, the percent of living cells went down to 54.2% with 3 μΜ gramicidin treatment compared with the control group. These data indicated that gramicidin promoted the cell apoptosis of SGC-7901cells in a time and dose dependent manner.
Fig. 3 Gramicidin induced apoptosis in SGC-7901 cells. SGC-7901 cells were treated with 0, 0.3, 1 and 3 μΜ of gramicidin respectively for a 24 h and c 48 h, and were then subjected to Annexin V-FITC/PI staining, followed by flow cytometer analysis. Quantification of the percentage of apoptotic cells which were treated with gramicidin at various doses after treatment for b 24 h and d 48 h. The results are shown as the mean ± SEM of three independent experiments (n = 3, *P < 0.05 and ***P < 0.001 vs. Control)
Gramicidin induced G2/M cell cycle arrest in human gastric cancer cells
We next explored the effect of gramicidin on the cell cycle of human gastric cancer cells. Propidium iodide (PI) staining was used to detect the cell cycle distribution, followed by flow cytometry. As shown in Fig. 4a, c, the percentage of SGC-7901 cells with gramicidin treatment in the G1 phase was lower, compared to the percentage of control group in the G1 phase in 24 h and 48 h. Whereas statistical analysis revealed that the population of cells in the G2/M phase was significantly increased in SGC-7901 cells in 24 h and 48 h (Fig. 4b, d). These results demonstrated that gramicidin could arrest cell cycle at G2/M phase.
Fig. 4 Gramicidin induced G2/M cycle arrest in SGC-7901 cells. Representative images of cell cycle distribution in a 24 h and c 48 h were shown. SGC-7901 cells were treated with 0, 0.3, 1 and 3 μM of gramicidin respectively. Quantification of the percentage of phase population (G1, S, G2/M) was showed in b and d. The results are shown as the mean ± SEM of three independent experiments (n = 3, *P < 0.05, **P < 0.01 and ***P < 0.001 vs. Control)
Gramicidin inhibited the migration of SGC-7901 cell
To detect the effect of gramicidin on migration ability of SGC-7901 cells, we next performed the wound healing assay. The data showed that the migration area of SGC-7901 with gramicidin treatment were obviously lower than the area of the control groups (Fig. 5a). Moreover, quantification analysis revealed that gramicidin resulted in a dose-dependent decrease of inhibiting the cell migration (Fig. 5b). Thus, the results suggested that gramicidin inhibited the SGC-7901 cell migration.
Fig. 5 Gramicidin inhibited the migration in SGC-7901 cells. a Typical images of wound healing analysis in SGC-7901 cells treated with indicated dosage of gramicidin were compared at 0 h and 48 h. b The cell mobility of SGC-7901 cells which were treated with 0, 0.3, 1 and 3 μM of gramicidin respectively for 48 h in 12-well plate was quantitatively analyzed by image J software. The results are shown as the mean ± SEM of three independent experiments (n = 3, *P < 0.05 vs. Control)
Gramicidin treatment downregulated the expression levels of Bcl-2, cyclin D1 and p-FoxO1
Next, we investigated the possible mechanism underlying the anti-cancer effect of gramicidin. The expression of apoptosis-related proteins was detected by western blot analysis. Figure 6a revealed a remarkable decrease in the expression of cyclin D1 and anti-apoptotic protein Bcl-2 compared with control group. In addition, the expression of p-FoxO1 was significantly downregulated following different concentrations of gramicidin. The fold change of Bcl-2 and cyclin D1 to β-actin and p-FoxO1 to total-FoxO1 were shown respectively in Fig. 6b-d, revealing that the regulatory effect of gramicidin on the expression of apoptosis-related proteins was in a dose-dependent manner.
Fig. 6 Gramicidin induced gastric cancer cell apoptosis by decreasing the expression of cyclin D1, Bcl-2 and p-FoxO1. a The expression of cyclin D1, Bcl-2 and p-FoxO1 in SGC-7901 cells which were treated with 0, 0.3, 1 and 3 μM of gramicidin respectively for 24 h were analyzed by western blot. β-actin was used as a loading control. The fold change of b Bcl-2 and c cyclin D1 to β-actin and d p-FoxO1 to total-FoxO1 were shown. The results are shown as the mean ± SEM of three independent experiments (n = 3, *P < 0.05, **P < 0.01 and ***P < 0.001 vs. Control)
The current study evaluated the cell cytotoxicity of gramicidin and its anti-proliferation activity in human gastric cancer cells. Meanwhile, the migratory capacity and G2/M cell cycle were inhibited upon gramicidin treatment with significant cell apoptosis. Furthermore, our results showed that this influence could be possibly attributed to the decreased expression of cyclin D1 and Bcl-2 with down-regulated FoxO1 phosphorylation. Despite of the recent findings in GC treatment which brought new targets and possible therapies like PD-1/PD-L1 checkpoint inhibitors [16], there is still no specific and effective drug that can cure gastric cancer. Thus, it is an urgent problem to find low toxicity and effective drugs to induce cell apoptosis while inhibiting cell proliferation and migration of tumor cells. Adenocarcinoma in gastric cancer has a high frequency of occurrence. Both SGC-7901 and BGC-823 were adenocarcinoma and these two types of gastric cancer cell lines were chosen as tools for biological studies. SGC-7901 was obtained from moderately differentiated lymph nodes of metastatic adenocarcinoma, while BGC-823 was obtained from undifferentiated adenocarcinoma. Their different sensitivities to gramicidin suggested that the level of differentiation may affect the anti-cancer gramicidin and SGC-7901 from metastatic adenocarcinoma lymph nodes with moderate differentiation was more sensitive to gramicidin. Although the effect of cell proliferation could not be excluded, we used low concentration of serum for migration assay. Thus, our study illustrated the effect of gramicidin on inhibiting cell proliferation, migration and apoptosis induction of gastric cancer cells while suggesting that gramicidin could be served as potential candidate for GC treatment.
Apoptosis is the typical kind of programmed cell death, which refers to the self-regulated and orderly death of cells controlled by multiple genes in order to maintain the stability of internal environment [17]. Resistance to cell death often leads to tumorigenesis while targeting apoptosis has been regarded as one of the common method in controlling cancer progression [18]. Among complex pathways regulating cell apoptotic fate such as ER, mitochondrial and TNF/TNF-R mediated cell death, mitochondria driven cell death plays a critical role [19, 20]. The process of mitochondria driven apoptosis involves the activation, expression and regulation of a series of genes and proteins like transcription factors and apoptosis-related Bcl-2 family. Bcl-2 proteins family, including anti-apoptotic Bcl-2 and pro-apoptotic BAX, regulates apoptosis by controlling mitochondrial permeability with subsequent release of caspases [20–22]. Correspondingly, our data also showed that Bcl-2 decreased at various doses of gramicidin which is consistent with the mitochondrial apoptosis pathway, suggesting that gramicidin might exert its pro-apoptotic effect through targeting mitochondria. Of note, the cell counting kit-8 assay showed significant toxicity of gramicidin on cell survival at 24 h while the expression of Annexin V did not altered significantly, suggesting that gramicidin might not only affect apoptosis but other form of cell death like autophagy.
Beside the cell apoptosis, targeting cell cycle has advanced the chemotherapy for decades. Dysregulated cell cycle would cause uncontrolled cell proliferation while permitting cells obtain the ability to replicate and grow without limitation [23, 24]. Thus, targeting cell cycle with inhibited cell replication could arrest neoplastic process [25]. In cell cycle movement, cyclins play critical roles in triggering and promoting cell cycle from G1 to S and G2/M phases. It was reported that cyclin D1 could be involved in diverse cycle status. Typically, the function of cyclin D1, or G1/S-specific cyclin-D1, is to promote cell proliferation by combining and activating the G1 period characteristic of the cycle dependence protein kinase CDK4 [26]. G1 phase cycle suppressor protein (Rb) is phosphorylated, and the phosphorylated Rb protein is dissociated from the E2F transcription factor to which it binds. E2F transcription factors initiate transcription of genes that are involved in the cell cycle, thus promoting the cell cycle from the G1 phase to the S phase [27]. However, in our study, little inhibition in G1/S were observed while higher inhibition in G2/M cell cycle were detected, suggesting that gramicidin might affect a different signaling to regulate the expression of cyclin D1 since cyclin D1 was also involved in signal transcription like Wnt/GSK-3 beta pathway in which cyclin D1 functions as a downstream factor to regulate cell survival [28]. Similarly, sweroside and lanatoside C could arrest leukemia cells or gastric cancer cells at G2/M while lowering the expression of cyclin D1 [29]. Our study further confirmed that gramicidin regulated cyclin D1 expression with G2/M inhibition and apoptosis, implying that modulating cyclin D1 might serve as a potential anti-tumor target.
FoxO1 is a transcriptional factor that plays a key role in the development and progression of gastric cancer. More studies have confirmed that FoxO1 is involved in a variety of cellular pathways, such as proliferation, stress resistance, differentiation and apoptosis. Many research focused on its regulatory effect on cell cycle. For example, polo-like kinase 1 (PLK1) interacts and phosphorylates FoxO1, abrogating its inhibitory effects on cell survival, thus promoting G2/M cycle progression [30]. In proliferating cells, FoxO1 could be activated by cyclin-dependent kinase 1 (Cdk1) to affect the expression of PLK [31]. In light of these findings, numerous research has proposed that regulating FoxO1 could be applied in cancer therapy. Knock down of FoxO1 inhibited tamoxifen sensitivity in breast cancer cells [32]. Our study revealed the effect of gramicidin on FoxO1 phosphorylation. After gramicidin treatment, the phosphorylation of FoxO1 decreased with the increase of cell apoptosis, indicating that increasing FoxO1 activity might contribute to cellular apoptotic fate.
Previous studies have shown that systemic intravenous or intraperitoneal injection of gramicidin is fatal to mice. However, repeated injection of gramicidin into the tumor can block tumor growth without obvious toxicity [15]. Toxicity limits the clinical application of gramicidin and many other pre-clinical drugs [33]. Systemic administration is generally considered to be a necessary means of treatment for invasive tumors. Local administration sometimes is an effective treatment for non-resectable or metastatic tumors and can be given by special means [34, 35]. With the development of science and technology, we believe that the toxicity of gramicidin can be solved via structure modification or combination usage with other clinical anticancer drugs, and this chemical can be applied as a novel drug or leading chemical for the treatment of GC in the future.
In summary, the antitumor effect of gramicidin on gastric cancer cells was preliminarily explored, in which gramicidin may inhibit cell proliferation and induce G2/M cell cycle block by down-regulating the FoxO1 and cyclinD1 while down-regulating tumor suppressor genes Bcl-2 to induce apoptosis. Combined with previous studies, gramicidin may have potential value as a clinical treatment drug for gastric cancer. The toxicity of drugs is one of the key problems to be solved. Meanwhile, the possible mechanism of its modulation of cell apoptosis merits further investigation.
This work was supported by grants from the The National Key Research and Development Program of China (2017YFC0908500), National Natural Science Foundation of China (81573438 and 81773741), Three-year Action Plan for Promoting Clinical Skills and Innovation in Municipal Hospitals (16CR4025A).
All data analyzed during this study are included in this article.
Forkhead box protein O transcription factor family
hypoxia-inducible factor
Von Hippel-Lindau
phosphate buffer solution
propidium iodide
Cdk1
We would like to thank the financial support of the grants from The National Key Research and Development Program of China (2017YFC0908500), National Natural Science Foundation of China (81573438 and 81773741), Three-year Action Plan for Promoting Clinical Skills and Innovation in Municipal Hospitals (16CR4025A).
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Recibido: 10 de Abril de 2019; Aprobado: 01 de Noviembre de 2019; : 25 de Noviembre de 2019
*Correspondence: ydhfighting@alumni.sjtu.edu.cn
fengqian@sjtu.edu.cn
wzshahbb@163.com
Tingting Chen and Yong Wang contributed equally to this paper
TTC, YW and YY performed the experiment. KKY, XLC, FC, HBX and YDP carried out data analysis and wrote the manuscript. YDH and FQ participated in preparing the manuscript. ZSW reviewed the manuscript. All authors read and approved the final manuscript.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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text:
Ann E. Ladd,
��������� Plaintiff-Appellant,
Robert G. Uecker and Milwaukee Brewers Baseball Club, L.P.,
��������� Defendants-Respondents.
On behalf of the plaintiff-appellant, the cause was submitted on the briefs of Ann Ladd, pro se.�
On behalf of the defendants-respondents, the cause was submitted on the brief of Katherine M. Longley of Foley & Lardner, LLP, Milwaukee.�
Cir. Ct. No.� 2008CV3169
Robert G. Uecker and Milwaukee Brewers Baseball Club, L. P.,
����������� APPEAL from an order of the circuit court for Waukesha County:� michael o. bohren, Judge.� Affirmed.�
�1������� SNYDER, J. Ann E. Ladd appeals the dismissal of her complaint and amended complaint against Robert G. Uecker and the Milwaukee Brewers Baseball Club, L.P.� The court granted Uecker�s and the Brewers� motion to dismiss because the court concluded that, as to most of the defamation and invasion of privacy claims, the complaint was filed beyond the statute of limitations, and the remaining allegations failed to state a claim because they involved statements protected by various privileges.� We agree and affirm.
�2������� Uecker is the radio broadcaster for the Brewers.� In June 2006, Uecker petitioned the Milwaukee County Circuit Court for an injunction against Ladd pursuant to Wis. Stat. � 813.125 (2007-08),[1] alleging a six- or seven-year pattern of harassment.� Around the same time, Ladd�a self-described �devoted fan��was charged with felony stalking.� The injunction petition hearing was held on July 3 and September 7, 2006.� The court commissioner found probable cause and issued an injunction.� Ladd did not seek de novo review or file a notice of appeal. �Soon after the injunction issued, the prosecutor dismissed the criminal charge.�
�3������� Without first giving written notice, see Wis. Stat. � 895.05(2), on September 8, 2008,[2] Ladd filed a sprawling pro se complaint alleging that between June 1 and September 7, 2006, Uecker defamed her in the affidavit supporting the injunction petition; he and/or the Brewers published the allegedly defamatory affidavit to a website called thesmokinggun.com; the Brewers posted on their website a defamatory article regarding her removal from a spring training game in Maryvale, Arizona; and a claim for �false light invasion of privacy� for, among other things, making and republishing false, defamatory statements and photographing her in the stands at various baseball stadiums.�
�4������� Uecker and the Brewers moved to dismiss Ladd�s complaint.� They asserted that: (1) the claims relating to Uecker�s affidavit are barred because they fall outside the Wis. Stat. � 893.57 two-year statute of limitations; (2) those claims are further barred by the absolute privilege for statements made in the course of judicial proceedings; (3) the sole defamation claim not barred by the statute of limitations, the March 2007 Associated Press (AP) news story appearing on the Brewers� website, is protected by the �wire-service� privilege; and (4) Wisconsin does not recognize �false light invasion of privacy� and Ladd pled no facts showing an invasion of any privacy rights Wisconsin does recognize.
�5������� In her response, Ladd acknowledged her �oversight� that Wisconsin does not recognize �false light invasion of privacy,� but contended that her complaint nonetheless stated an invasion of privacy cause of action under Wis. Stat. � 995.50.� Four days before the hearing on the motion, Ladd filed an amended complaint materially the same as the original complaint except that she renamed the prior �False Light Invasion of Privacy�Distress� to �Emotional Distress and Invasion of Privacy.�
�6������� After a hearing, the circuit court concluded that the statute of limitations bars Ladd�s claims for all matters occurring before September 7, 2006; that statements Uecker made during the September 7, 2006 hearing enjoy absolute judicial immunity; that the AP news story is protected by the wire-service privilege; and that Ladd�s invasion of privacy claims lack legal support.� Based on Ladd�s assertion that the amended complaint essentially mirrored the initial one, the court dismissed them with prejudice.� Ladd filed this appeal.
�7������� A motion to dismiss tests the legal sufficiency of the complaint.� Doe v. Archdiocese of Milwaukee, 211 Wis. 2d 312, 331, 565 N.W.2d 94 (1997).� On review, we accept as true the alleged facts and their reasonable inferences, but we draw all legal conclusions independently.� Walberg v. St. Francis Home, Inc., 2005 WI 64, �6, 281 Wis. 2d 99, 697 N.W.2d 36.� When a plaintiff clearly cannot recover under any conditions, a motion to dismiss should be granted.� Meyers v. Bayer AG, 2006 WI App 102, �7, 293 Wis. 2d 770, 718 N.W.2d 251, aff�d, 2007 WI 99, 303 Wis. 2d 295, 735 N.W.2d 448.
�8������� The elements of a common law action for defamation are: (1) a false statement; (2) communicated by speech, conduct or in writing to a person other than the one defamed; and (3) the communication is unprivileged and tends to harm one�s reputation, lowering him or her in the estimation of the community or deterring third persons from associating or dealing with him or her.� Torgerson v. Journal/Sentinel, Inc., 210 Wis. 2d 524, 534, 563 N.W.2d 472 (1997).� If we determine the matter complained of is not defamatory, that generally ends the matter.� See Lathan v. Journal Co., 30 Wis. 2d 146, 151, 140 N.W.2d 417 (1966).[3]� If it is defamatory, we must consider the defenses alleged.� Id.� Truth is a complete defense.� Id.� Even if false, however, it still may have the benefit of either absolute or conditional privilege.� See id.� An absolute privilege gives complete protection while a conditional privilege may be forfeited if abused.� Vultaggio v. Yasko, 215 Wis. 2d 326, 331, 572 N.W.2d 450 (1998).
�9������� Ladd�s September 8, 2008 complaint alleges that Uecker defamed her:� (1) in the affidavit in support of his petition for the harassment injunction;
(2) by publishing the affidavit to thesmokinggun.com; (3) during the two-day injunction hearing; and (4) in a media interview after the first day of the hearing.� Distilled to its essence, Ladd�s claim is that the false depiction of her as a stalker has damaged her personal and professional reputations.� Except for the continued injunction hearing on September 7, 2006, however, all of these incidents occurred more than two years before Ladd filed her complaint.� An action to recover damages for a defamatory communication is barred if not commenced within two years after the cause of action accrues.� Wis. Stat. � 893.57.
�10����� Ladd disagrees.� She argues that the statute of limitations does not bar her claim because, under Hansen v. A.H. Robins, Inc., 113 Wis. 2d 550, 560, 335 N.W.2d 578 (1983), her defamation claim did not accrue until she discovered or reasonably could have discovered her injury. �Neither Ladd�s original nor her amended complaint contains any factual allegations implicating the discovery rule, however. �This assertion has no merit.
�11����� Ladd also argues that, although Uecker and/or the Brewers allegedly posted his affidavit to thesmokinggun.com on June 2, 2006, the purportedly defamatory statements still can be accessed on the Internet today.� She contends that the information therefore is republished each time someone visits that website or others to which the material has found its way, thus renewing her cause of action.� We disagree.� �Any one edition of a book or newspaper, or any one radio or television broadcast, exhibition of a motion picture or similar aggregate communication is a single publication.�� Restatement (Second) of Torts
� 577A(3) (1977).� Wisconsin has not yet addressed the multiple- or single-publication rule head-on.� See Voit v. Madison Newspapers, Inc., 116
Wis. 2d 217, 227, 341 N.W.2d 693 (1984) (Abrahamson, J., concurring); see also id. at 223.� The great majority of courts have, however, followed the single-publication rule. �See Keeton v. Hustler Magazine, Inc., 465 U.S. 770, 777 n. 8 (1984); see also Restatement (Second) of Torts � 577A, Reporter�s Note.
�12����� Some courts have applied the single-publication rule specifically to publication on the Internet.� See, e.g., Firth v. State, 775 N.E.2d 463, 465-66 (N.Y. 2002).� We do so here.� We hold that �republishing� the allegedly defamatory information about Ladd on the Internet is not actionable.� Accepting as we must on this review that Uecker or the Brewers were responsible for the initial publication to thesmokinggun.com on June 2, 2006, that act is outside the statute of limitations.[4]� Uecker and the Brewers have no control over other websites� use or dissemination of the same information on the World Wide Web. �We reject the notion that each �hit� or viewing of the information should be considered a new publication that retriggers the statute of limitations.
�13����� As to Uecker�s September 7, 2006 injunction hearing testimony, even if it conceivably could be construed as defamatory, it is not actionable.� Statements �pertinent or relevant to the case� that are made in the course of judicial proceedings are absolutely privileged and insulate the speaker from liability.� Bergman v. Hupy, 64 Wis. 2d 747, 750, 221 N.W.2d 898 (1974).� Uecker�s written affidavits and courtroom testimony are absolutely privileged.
�14����� Ladd asserts, however, that Uecker�s statements lost their absolute privilege through �excessive publication� on the Internet, because the �stalker label� �defame[ed] [her] as a criminal� and because Uecker defamed her to law enforcement officials.� See Restatement (Second) of Torts � 604 (1977) (addressing loss of privilege through excessive publication of defamatory material); State v. Gilles, 173 Wis. 2d 101, 111, 496 N.W.2d 133 (Ct. App. 1992) (stating that a conditional privilege applies to criminal defamation); and Heggy v. Grutzner, 156 Wis. 2d 186, 192, 456 N.W.2d 845 (Ct. App. 1990) (stating that defamatory statements to law enforcement officers are conditionally privileged).� Ladd is mistaken in several respects.
�15����� First, we are unpersuaded that Uecker�s factual statements amount to defamation in the first instance, despite negative fallout to Ladd.� Second, even if untrue, statements made in judicial proceedings are absolutely privileged.� Lathan, 30 Wis. 2d at 151.� An absolute privilege gives �complete protection.�� Vultaggio, 215 Wis. 2d at 331.� Third, these were civil, not criminal, proceedings; even so, the absolute privilege for statements made in judicial proceedings applies equally to criminal and civil defamation.� See State v. Cardenas-Hernandez, 219 Wis. 2d 516, 541, 579 N.W.2d 678 (1998).� Fourth, Uecker�s June 2006 statements to the police in connection with his attempt to secure an injunction against her are beyond the statute of limitations.� Finally, the statute of limitations also defeats Ladd�s �excessive publication� claim regarding Uecker�s June 2006 affidavit.
�16����� Ladd�s complaints that the Brewers defamed her likewise fail.� The Brewers advised Ladd in December 2006 that, in light of the harassment injunction, they would deny her entrance to the spring training facility in March 2007 should she purchase a ticket.� Upon finding her in the stands, they were entitled to have her removed.� As Ladd�s ticket indicates,[5] a ticket of admission to a place of amusement is simply a license to view a performance that the owner or proprietor may revoke at will.� See 27A Am.Jur. 2d Entertainment and Sports Law � 42 (2008); see also Soderholm v. Chicago Nat�l League Ball Club, Inc., 587 N.E.2d 517, 520-21 (Ill. App. Ct. 1992).� Any statements the Brewers may have made to the Phoenix police in connection with the incident enjoy a conditional privilege, see Heggy, 156 Wis. 2d at 192, which Ladd has not rebutted by showing actual malice.� See Otten v. Schutt, 15 Wis. 2d 497, 504, 113 N.W.2d 152 (1962).�
�17����� Ladd then directs us to an allegedly defamatory March 20, 2007 article in the Brewers� online news archive about the Maryvale incident.� Assuming, as Ladd contends, that the Brewers posted the story there, and accepting simply for argument�s sake that the article is defamatory, this claim also fails.� Before filing suit, Ladd did not give written notice to the Brewers providing them �a reasonable opportunity to correct the libelous matter.�� See Wis. Stat. � 895.05(2).� Her failure to give notice is fatal to her claim.� See Hucko v. Jos. Schlitz Brewing Co., 100 Wis. 2d 372, 374, 302 N.W.2d 68 (Ct. App. 1981).
�18����� Furthermore, the report bears an AP copyright.� The Brewers are protected by the �wire-service� privilege, under which news disseminators �that rely on the accuracy of a wire service release are not negligent as a matter of law.�� See Van Straten v. Milwaukee Journal Newspaper-Publisher, 151 Wis. 2d 905, 920, 447 N.W.2d 105 (Ct. App. 1989) (describing the privilege); see also Cole v. Star Tribune, 581 N.W.2d 364, 368 (Minn. Ct. App. 1998) (stating that Wisconsin has adopted it).� The AP story also bears a disclaimer that it �was not subject to the approval of Major League Baseball or its clubs.��
�19����� Lastly, Ladd acknowledges that Wisconsin does not recognize false light invasion of privacy, to which she made an �errant� reference in her initial complaint.� She argues, however, that Uecker and the Brewers nonetheless invaded her privacy by recklessly or unreasonably disclosing her private information, which caused her to suffer �massive repercussions on [her] livelihood and well-being.�
�20����� In Wisconsin, �invasion of privacy� means a highly offensive intrusion upon another�s privacy in a place a reasonable person would consider private or in a manner actionable for trespass; using, without written consent, a person�s name, portrait or picture for advertising or trade; giving publicity to a person�s private matters �of a kind highly offensive to a reasonable person,� unless the information is available to the public as a matter of public record; or conduct involving depictions of nudity.� Wis. Stat. � 995.50(2).� Ladd alleges that the Brewers took photographs of her in the stands at baseball parks and disseminated her �mug shot� and information about the injunction and the spring training incident.� None of these involved private places, using her likeness for advertising or trade, or depictions of nudity.� Further, they are matters of public record.� She did not plead facts that satisfy an invasion of privacy claim.
[1] All references to the Wisconsin Statutes are to the 2007-08 version unless noted.
[2] September 7, 2008, fell on a Sunday, giving Ladd until September 8.� See Wis. Stat. � 990.001(4)(b).
[3] We opt to address all of Ladd�s claims on the merits.
[4] Even apart from the single-publication rule analysis, the thesmokinggun.com posting updated in October 2006 is not actionable.� It contains only verifiable, objective facts regarding the issuance of the injunction and dismissal of the felony stalking charge against her.� Truth is a complete defense.� Lathan v. Journal Co., 30 Wis. 2d 146, 151, 140 N.W.2d 417 (1966).
[5] Ladd included a photocopy of her ticket as an exhibit, evidently to show she had a right to be at the game.� The ticket reads:� �The license granted by this ticket to enter the Club baseball game is revocable.��


text:
by Kevin Helms
Korean Regulators Widen Investigation of Cryptocurrency Exchanges
South Korea's government is widening its probe on cryptocurrency exchanges, particularly the use of corporate accounts which the regulators say can lead to money laundering. This announcement follows the prosecutors launching an investigation on the country's largest crypto exchange, Upbit.
Also read: Yahoo! Japan Confirms Entrance Into the Crypto Space
Widening the Crypto Probe
South Korea's top financial regulators are teaming up with prosecutors to widen their investigation of domestic cryptocurrency exchange operators. An official of the Financial Service Commission (FSC) was quoted by the Korea Times on Sunday:
Following a request by the Financial Supervisory Service (FSS) and the prosecution to address growing anti-money laundering compliance concerns and possible abuse of cryptocurrencies in money laundering and fraud, the FSC is looking into exchanges' corporate accounts opened in local banks.
The use of corporate accounts for crypto transactions should have been discontinued when the government introduced the real-name system at the end of January. However, only 30% of all crypto accounts have been converted into real-name ones so far.
The six banks that have the ability to issue real-name accounts have chosen to only service the country's largest crypto exchanges: Upbit, Bithumb, Coinone, and Korbit. Nonetheless, not all accounts at these exchanges have been converted into real-name ones. In addition, all small and medium-sized exchanges continue to use corporate accounts for crypto transactions.
The regulators say that the use of corporate accounts can lead to fraud such as recently seen with Coinnest whose CEO was charged with embezzlement. The largest Korean crypto exchange by volume, Upbit, is also currently under investigation even though banks have been converting its accounts to real-name ones.
Collaborating with Other Countries
South Korea has been discussing a collaboration with other countries on cryptocurrency regulations. At a recent International Organization of Securities Commissions (IOSCO) Board of Directors and Annual General Meeting held in Hungary, FSC Vice Chairman Kim Yong-beom discussed cryptocurrency issues with major national supervisory bodies. The need for IOSCO to cooperate on cryptocurrency and ICO regulations was stressed at the meeting.
An official of the FSC was quoted by the Korea Times saying:
The FSC is collaborating with authorities in other countries. Our latest findings show that the domestic exchange faked its balance sheets and deceived investors. The FSC is checking Upbit's computer system with prosecutors and the FSS to audit the exchange's virtual currency holdings.
Meanwhile, the new FSS governor Yoon Suk-heun recently indicated that he will look into easing regulations on domestic cryptocurrency trading, citing that "there are some positive aspects to cryptocurrencies."
What do you think of the Korean regulators widening probe on crypto exchanges? Let us know in the comments section below.
Images courtesy of Shutterstock and the Korean government.
banks, Bitcoin, Bithumb, BTC, Coinone, Cryptocurrency, Digital Currency, Exchanges, FSC, FSS, Korbit, korea, korean, Money Laundering, N-Economy, South Korea, upbit, virtual accounts, Virtual Currency
Need to calculate your bitcoin holdings? Check our tools section.
Kevin Helms
A student of Austrian Economics, Kevin found Bitcoin in 2011 and has been an evangelist ever since. His interests lie in Bitcoin security, open-source systems, network effects and the intersection between economics and cryptography.
Pakistan Telecom Authority Asked to Block Over 1,600 Crypto Sites — Minister Says Government Intends to 'Regularize' Crypto
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Here's What Happens When You Click on a Spammy Cryptocurrency Ad
Ethereum Futures in US One Step Closer as CME Deal is Struck
A group of U.S.-based banks is launching their own stablecoin, USDF. The stablecoin will be issued by the USDF Consortium, which will allow its members (financial and banking institutions) to issue USDF. The proposed stablecoin will be the first currency ... read more.


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June 9, 2017 by zekezielinski
Legal beagle says POTUS has the right to decide who to investigate and who to stop investigating
James Comey says President Trump is a liar.
President Trump says James Comey is a liar.
Liar, liar, pants on fire!
Lawdy, I'm so glad George Washington never told a lie.
Now here's something worth chewing on like gristle: Perhaps Trump had every right to fire Comey at will and to directly tell him to lay off Michael Flynn and stop investigating whether there was any collusion between the Trump campaign and our dear Russian friends.
Who says so?
Not me. Legal mumbo jumbo is all Latin to me even when it is in some English dialect.
Alan Dershowitz says so.
Dershowitz, Felix Frankfurter Professor of Law Emeritus at Harvard Law School who is a prominent scholar on United States constitutional law and criminal law and a leading defender of civil liberties, says the president has the authority to direct the FBI to stop investigating any individual and can, in theory, decide who to investigate, who to stop investigating, who to prosecute and who not to prosecute. The president is the head of the unified executive branch of government, and the Justice Department and the FBI work under him and he may order them to do what he wishes.
According to Dershowitz, who was a member of the famous (or infamous) O.J. Simpson legal dream team, the president had the right to order Comey to pogo stick down to Mar-a-Lago or have the FBI investigate why the wind messes with Trump's comb-over.
Trump is more of a provocateur than a president
President Trump reignited feuds with the mayor of London, the media, Democrats and his own Justice Department with a new series of tweets Monday that begins a critical week for his administration, one that will include former FBI director James Comey testifying to the Senate Intelligence Committee.
Who's next? Mickey Mouse? Elsa from Frozen?
The man obviously loves to pick a fight. His nervous system mimes rage so well that it becomes real, much like an actor's tears.
Twitter, as well as the Russians, may unravel the Trump presidency like cheap rayon.
Trump, by the at way, has opted not to invoke executive privilege and won't try to block Comey's testimony.
Which means he is not always as dumb as an ash tray (remember them?).


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Territory Stories
Submission to the Australian Government Review of Remote Employment Services
Central Land Council annual report; Central Land Council reports; Reports; PublicationNT
Made available via the Publications (Legal Deposit) Act 2004 (NT).
Central Land Council (Australia) -- Periodicals; Aboriginal Australians -- Northern Territory -- Land tenure -- Periodicals
Central Land Council
Copyright owner
Check within Publication or with content Publisher.
Parent handle
https://hdl.handle.net/10070/260449
Citation address
4 work history. Often basic literacy and numeracy skills are identified by new Rangers as their number one priority. The CLC has found that it is also important to understand and recognise the extent of variation from one region to another where Ranger groups operate. There are strong historical differences between regions. For example in Docker River there has never been any sort of structured work, which appears to have lead to a situation where there is no strong culture of mainstream work. This contrasts, for example, Hermannsburg and Santa Teresa where there is a mission history and the legacy of work building up the pastoral industry. Evaluating Ranger performance in Docker River and Hermannsburg requires different factors to be taken in to account. This is not about having lower expectations but rather understanding the context and how it comes to bear on the individuals work readiness. Then there are also variables within Ranger groups. Individual work histories, or a lack thereof; levels of literacy and numeracy; impacts of substance abuse and physical health are examples. Without knowing people personally and understanding all of that background and having a relationship the development doesnt come. A good example where there is a tailored approach which combines capacity with opportunity is the Tjuwanpa Outstation Resource Agency. Tjuwanpas operations are based on an high degree of understanding of the local context and there is strong community support. In this example the Tjuwanpa training centre is located next to the Tjuwanpa Ranger Office; where there are real jobs. There is a strong relationship between the CLC Ranger Coordinator and the training centre coordinator on the ground. They communicate with one another such that the training coordinator understands the direct training that people would need to move in to the Ranger program. Some people have moved across to work as Rangers. Through this level of collaboration the relationship promotes success. Community support and alignment of shared values The communities in which Ranger programs operate support the programs and view the positions as real and legitimate jobs. This is a crucial element in their success. Much of the appeal of work as a Ranger is that people can see it is success on their own terms. They see from other Ranger groups that Rangers arent just passive participants in a program. The programs are also community driven. Crucially, there is input and support from Aboriginal traditional landowners in developing work plans and incorporating traditional knowledge. This is an example of where values are shared and aligned. In this example, the mainstream value of gainful waged employment aligns with a relationship to country, the imperative of traditional knowledge transfer and a custodial relationship with the land. There is an intergenerational aspect to work that aligns shared values and enjoys strong community support. Young kids see their parents and members of their community turning up to work week in week out. This has a strong psychological effect and creates role models.
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Our landscaper put in four pieris plants in a terraced planter on the west side of our new home two years ago. The first year they were beautiful but they have been declining since then. New leaves come out but turn brown starting at the tips. My husband says they just need more water although I have given them copious amounts. I think they receive too much sun to survive. Is there anything I can do for them?
Pieris is a wonderful shrub for Arkansas if it is growing in the shade--not the west side of your home. I too think they are a bit unhappy with the amount of sunlight they are getting, but I would also look at the drainage. Pieris plants won't tolerate wet feet. If the soil is heavy, they may be retaining too much moisture. Plants can wilt from a lack of moisture or too much. Dig a hole next to the plants--away from the root system, and see how wet or dry the site is. Consider moving them this fall to a location with morning sun and afternoon shade, and plant them on the high side to encourage drainage.


Upon opening the email, I was greeted with a series of comparative photographs — some color photos showing Democratic presidential contender and U.S. Sen. Elizabeth Warren of Massachusetts visiting with Mississippi Delta residents in Cleveland in 2019 alongside black-and-white photographs of the late U.S. Sen. Bobby Kennedy visiting Delta residents in 1967.
The attempt to compare Warren's 2019 visit to the Delta with Kennedy's 1967 visit seems exceedingly shallow and contrived — in a way almost politically and morally blasphemous. First and foremost, comparing the state and nation's political and social climate in 1967 with that of today is telling.
There was true danger confronting all concerned when Kennedy made his pilgrimage to Mississippi in 1967 to see the poverty he'd heard Senate committee testimony about from activist Marian Wright. Kennedy and those traveling with him risked being targeted for violence in the Mississippi that existed in that era.
Likewise, the impoverished African-American Delta families who allowed their homes to be used as backdrops for Kennedy's visit faced perhaps even more raw danger from drawing attention to Mississippi's inequalities from white supremacists. When RFK visited the Mississippi Delta, it was considered by many Mississippians as a political provocation rather than a humanitarian mission.
But the fact is that federal government efforts to deal with poverty and hunger in America were still in their practical infancy. Food stamps began as a pilot project in 1961 during the administration of President John F. Kennedy with a dual purpose — to feed the hungry, yes, but also to provide a destination for surplus farm commodities.
RFK's visit indeed did bring a greatly heightened level of national awareness to poverty and hunger in the Mississippi Delta and in the Appalachian South, but by October 1967 national food stamp use was only at the 2 million mark. Still led in that day primarily by segregationist elected officials, Mississippi was a very reluctant participant in the food stamp program. But by fiscal year 2015 in Mississippi alone, food stamps, or the Supplemental Nutrition Assistance Program (SNAP), provided $920 million in benefits to a monthly average of 636,322 people.
Federal dollars make up about 42 percent of all state revenue in Mississippi, and federal transfer payment account for 26.3 percent of all personal income in the state. Does Sen. Warren seriously think that in a nation where Social Security and Medicare are already in mortal fiscal danger, the Mississippi Delta's poverty can be cured by more social programs?
And how to pay for those promised new programs? Warren pitched her "wealth tax" plan — the so-called "net worth" tax. Perhaps nothing about her 2019 visit to Mississippi was more discordant with the Kennedy's philosophies than that.


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More than 30 edible gift ideas.
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Last minute gift idea: Movie Tickets.
Easy DIY wrapping paper with Bic Mark-It markers.


The European Central Bank was once known for its focus on price stability. Since the global economic crisis, however, its role has extended to saving banks and sovereign countries. This column argues that such a move has badly harmed the institution's legitimacy – something that will damage both its policy effectiveness and confidence in the governing bodies of the EU as a whole.
The ECB's role has evolved in its decade-long existence. In this note I describe how the choices of the ECB have damaged the institution's legitimacy. This matters because decreased legitimacy lowers the ECB's future policy effectiveness and weakens the legitimacy of all other institutions of governance in the EU, including the European Commission, the European Court of Justice, and the European Parliament.
The period after when the ECB was called upon to help contain the liquidity crisis and, within its powers as a central bank, to mitigate the banking sector insolvency and sovereign debt crises.
Prior to August 2007 the ECB produced stable and only slightly above-target inflation. However, it lacked procedural transparency. The Treaty makes it clear that it was intended that members of the Governing Council should vote. An astonishing 834 words are devoted to describing exactly how the voting mechanism is to work. Unfortunately, in comparison to other central banks that are supposed to make decisions by majority rule, such as the central banks of Japan, Sweden, the UK, and the US, the ECB is opaque about how its policy rate decisions are reached. It does not reveal the vote or publish minutes. When questioned by the press or the European Parliament about the lack of procedural transparency, ECB President Trichet's stock response has been to view transparency as being equivalent to explaining one's decisions ex post and to extoll the virtue of the ECB in this regard.
It turns out that votes on policy rates are never taken. Apparently, some small subset of the Governing Council decides, prior to the meeting, what the policy rate will be and this is then presented to the entire Governing Council, which we are to believe always or almost always unanimously approves. This explains how the Governing Council is able to produce the lengthy post-policy meeting statement that Mr Trichet views as the ECB's major contribution to transparency. With the decision made before the meeting, there is ample time to prepare it.
That this extraordinary decision-making mechanism has gone on for so long with no formal explanation beggars belief. Who gets to make the decision? Why is it that no Governing Council member has ever insisted upon their legal right to a vote on monetary policy? Is there really never any dissent? How can that be? That we are able to ask these questions about an institution that is supposed to be one of the world's two most important central banks is not good for its legitimacy. Moreover, while this arrangement has functioned well enough so far, what would happen if some future president were a little less like Mr Trichet and a little more like, say, Davíð Oddsson?
In Mr Trichet and the Governing Council's defence, the ECB is woefully badly designed. The Governing Council has 23 members – a ludicrous size for a decision-making body. If each member gets a ten-minute opening statement, the rate-setting meeting would have gone on for almost four hours before any actual debate begins. With no formal way of reducing the size of the decision-making body, Mr Trichet may have had little choice but to make monetary policy informally.
In the period after the crisis the ECB has done a reasonable job relative to other central banks. It initially responded to the August 2007 liquidity crisis by dousing markets with liquidity in an attempt to drive down the interest rate. This was bad as the problem was not that financial institutions could not borrow at a reasonable interest rate using the good-quality collateral demanded by the Eurosystem. Instead, it was that they could not sell their asset-backed securities or use them as collateral because the markets for these assets had dried up. However, as the liquidity crisis continued and widened into a solvency crisis, the ECB's policies evolved. When – for example – the spread between the three-month LIBOR and the overnight indexed swap rate widened, the ECB responded sensibly by undertaking liquidity-providing longer-term refinancing operations with a maturity of three months and offsetting these with their main (short-term) refinancing operations. In mid-October 2008 it made the necessary and massive changes in what was to constitute eligible collateral.
Unfortunately, the ECB has not been a model of openness in its financial stability role. It does not tell us how it values illiquid marketable securities or how it decides its haircuts. When a member of the ECON committee of the European Parliament asked about how asset-backed securities are valued for collateral, Trichet said, "This is done by the system in ways which I considered appropriate but that we can improve at any time if we judge that they should be improved…". In other words, I'm not going to tell you and only our opinion matters.
The ECB's haircut policies are hard to fathom. Why do haircuts increase sharply with an asset's maturity even when the potential illiquidity does not necessarily do so? This policy only encourages the issuance of short-term debt. No wonder banks are facing massive short-term debt refinancing requirements during a period that they are also trying to access the markets for additional capital. With no explanation, one might wonder if the ECB even has a coherent theory of how to determine haircuts.
This opacity about collateral policies is potentially much more damaging to legitimacy than the lack of transparency in monetary policy. In deciding what securities to accept as collateral and how to value and haircut them, the Eurosystem is redistributing wealth.
In its attempt to maintain financial stability the ECB and Eurosystem have had to walk a fine line between providing just enough liquidity to keep potentially solvent institutions afloat and subsidising the financial sector. Given the lack of transparency it is not easy to judge how well they have done at this, but a couple of examples show that sometimes – through design or otherwise – they have strayed into the subsidisation territory.
One example was the policy of allowing Icelandic banks to borrow from the Eurosystem using each other's debt as collateral.
Another was the unlimited one-year fixed rate liquidity provision of June 2009 which may have been a transfer of about €1 billion from taxpayers to delighted banks (see Buiter 2009).
Playing Santa Claus to banks – Icelandic or otherwise – is not part of the ECB's mandate and does not enhance its legitimacy.
So far, the ECB appears to have been less competent in managing the sovereign debt crisis than it was in managing the liquidity and banking crises. That ECB policymakers did not speak out about the state of Greek finances, although they must have realised by 2004 that Greece was verging on insolvency, and that they continued to accept Greek sovereign debt on the same terms as they accepted German sovereign debt must have suggested to the market that there were two possibilities. Either, the ECB had superior information suggesting Greece was unlikely to be insolvent or the ECB knew that Greece – or at least its creditors – would be bailed out if Greece were to become insolvent. This may have led the market to be complacent for far too long.
The introduction of the Securities Market Programme threatens the ECB's legitimacy as the potential fiscal role is an inappropriate activity for an independent central bank. One can imagine that it must have been distasteful to Mr Trichet to undertake such a task, but – understandably – he may have viewed it as a less unpalatable option then allowing Greece to default in May 2010.
Not all uses of the Securities Market Programme are inappropriate, however.
In the event of a wholesale creditor run based solely on self-fulfilling expectations, it is reasonable for a central bank to intervene and act as a lender of last resort to financial institutions that would otherwise be solvent.
Likewise, in the event of a run on sovereign debt that is based solely on self-fulfilling expectations, it is reasonable for a central bank to make it clear that it stands ready to purchase as much of the debt as it takes to halt the run.
As the recent rise in the yields on Spanish and Italian debt was likely a result of fear-based self-fulfilling expectations, the ECB was justified in its purchases of this debt.
The failing of the institution is that it is not credible that it is willing to purchase enough of the debt to contain the run.
Unfortunately, the ECB's insistence on secrecy with respect to the programme is particularly damaging. It is widely believed that the average discount to face value paid for the Greek debt acquired (prior to August at least) was no more than 20%. The national central banks appear to have sought out the lucky counterparties. And, the ECB won't say who they are or how much they paid. The possibilities for corruption under such a system are endless. And one does not have to be a conspiracy theorist to imagine that taxpayers throughout the Eurozone were called upon to subsidise German and French banks after their own governments failed to exercise proper regulation and supervision.
The incoming ECB president does not face an enviable situation. The sovereign debt crisis is gaining strength, dampening the economic recovery that might have otherwise been expected to follow the liquidity and solvency crises. His predecessor set a reasonably high standard for competency, but his opacity damaged the legitimacy of the organisation.
Buiter, Willem (2009), "Recapitalising the Banks through Enhanced Credit Support: Quasi-Fiscal Shenanigans in Frankfurt", Maverecon, Financial Times, 28 June.


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P ortuguese Heritage Publications of California will make available, free of charge, the book "America" to any organization or individual preparing Portuguese speaking immigrants for their citizenship test.
This hard cover book, in the Portuguese language, not only contains the history of the United States but also a complete translation of many of its most important documents. In addition, it includes a sample of the citizenship test.
"America" is a valuable tool for anyone wishing to acquire her/his American citizenship. PHPC is happy to make the book available to organizations or individuals preparing others for citizenship. To request a book for your organization, please leave a comment on this blog with the organization's name and shipping address.


Meditation isn't about sitting still and doing nothing. It's about becoming a more authentic person, a new person, or even a better person. It's about going inward, increasing your awareness, and getting a healthy sense of perspective. You're not trying to turn off your thoughts or feelings. You're learning to observe them without judgment. And eventually, you may start to better understand them as well.


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Bill Clinton Hints That Hillary Could Run Again - The Political Insider
Bill Clinton Hints That Hillary Could Run Again
Bill Clinton hinted that his wife Hillary could run for office again in the future, further fueling speculation that she may jump into the 2020 Democrat primary.
The former President made the comments as he joined his wife and Supreme Court Justice Ruth Bader Ginsburg for a public 'conversation' at Georgetown University in Washington on Wednesday.
Rumors have been swirling amongst top aides for the former First Couple and pundits alike for weeks, and Clinton's comments did nothing to quell speculation.
"She may or not ever run for anything," Clinton said pointing at two-time failed presidential candidate, Hillary. "But I can't legally run for President again."
RELATED: Former Clinton Adviser: Hillary Thinks God Put Her On Earth to be President
"She may or not ever run for anything, but I can't legally run for President again." – Bill Clinton leaving the door open on @HillaryClinton running for President in 2020. #StillWithHer #Hillary2020 pic.twitter.com/gDrmcR0ax1
— Joel McAuliffe (@JdMcAuliffe) October 31, 2019
May or May Not Run
A Washington Post article about the conversation notes that, as per usual, Hillary made the discussion session, which was geared toward reminiscing about Ginsburg's nomination and tenure on the court, political and all about her.
She complained about the number of federal judges President Trump is appointing "being pushed through despite having no relevant experience."
She claimed it was her voice that helped Bill choose Ginsburg, stating "I may have expressed an opinion or two about people he should move up."
She insisted she completed the "RBG workout," a famed strenuous exercise routine for the 86-year-old Justice, but chose not to have cameras present.
She wouldn't commit to choosing sides in the World Series because, as Clinton said according to the Post, "she might still run for something someday."
RELATED: Former Top Hillary Aide: She Hasn't Ruled Out 2020 Run
A former Bill Clinton adviser says Hillary believes "God put her on the Earth" to be President. pic.twitter.com/epfg3xkAQV
— Fox & Friends First (@FoxFriendsFirst) October 28, 2019
Will She or Won't She?
Bill Clinton's non-committal statement in closing the door on his wife's future political prospects is another in a long list of examples of people associated with the couple suggesting she may run – if the conditions are right.
Just last week, Philippe Reines, a former top aide to Hillary Clinton, says his old boss had not ruled out running in 2020.
"If she thought she had the best odds of beating Donald Trump, I think she would think about it long and hard," Reines assessed.
RELATED: Hillary Can't Help But Attack Trump at Elijah Cummings Funeral
.@HillaryClinton on Rep. Elijah Cummings: "Like that Old Testament prophet, he stood against corrupt leadership of King Ahab and Queen Jezebel."
Full video here: https://t.co/3Nt7BSmMFq pic.twitter.com/lnHZvVENKC
— CSPAN (@cspan) October 25, 2019
Dick Morris, a former adviser to President Bill Clinton in the 1990s, said earlier this week that Hillary believes "God put her on the Earth" to be president.
Morris also indicated that she'd most likely enter the fray if former Vice President Joe Biden faltered.
"My feeling is that she wants to. She feels entitled to do it. She feels compelled to do it. She feels that God put her on the Earth to do it. But she's hesitant because she realizes the timing is bad," the former Clinton ally explained. "She's got to wait until Biden drops out because he's obviously next in line for it, and if he goes away, there's an opening for her."
And Hillary herself has done nothing to tamp down the rumors. After President Trump taunted her with getting into the 2020 race, she responded, "Don't tempt me."
Hillary later added, "I can beat him again."
President Trump Shows His Support for Nick Sandmann's Lawsuit Against Washington Post


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Friedrich IV. von Sachsen-Gotha-Altenburg (* 28. November 1774 in Gotha; † 11. Februar 1825 ebenda) war der letzte Herzog von Sachsen-Gotha-Altenburg.

Leben

Friedrich IV. war der dritte Sohn des Herzogs Ernst II. von Sachsen-Gotha-Altenburg (1745–1804) aus dessen Ehe mit Charlotte (1751–1827), Tochter des Herzogs Anton Ulrich von Sachsen-Meiningen. Zwischen 1788 und 1790 wurde er gemeinsam mit seinem älteren Bruder August in Genf ausgebildet. Friedrich studierte Philosophie, Staatsrecht und Geschichte. Im Gegensatz zu seinem Bruder galt er als bescheiden, gutmütig und volksnah. Der Prinz war ein bewunderter Sänger.

Von seinem Vater war Friedrich als jüngerer Sohn für den Militärdienst bestimmt und 1785 als Oberst in die niederländische Armee gegeben worden. Er kämpfte in den Feldzügen gegen das revolutionäre Frankreich und wurde 1793 durch einen umstürzenden Pulverwagen schwer verwundet. Infolge dieser Verwundung zog er sich ein Leiden zu, das ihn zwang, sich zeit seines Lebens viel in Heilbädern aufzuhalten und an dem er schließlich später starb.

Von 1804 bis 1810 lebte Friedrich in Rom. Von Papst Pius VII. wurde ihm ein Miniatur-Obelisk geschenkt; 1814 konvertierte er als einziger in der Geschichte seines Hauses in Italien zur römisch-katholischen Kirche. Der Aufenthalt in Rom hatte seinen Gesundheitszustand deutlich verbessert.

Friedrich hatte ein starkes Interesse an Ägypten und dem Orient; er war Hauptförderer der Orientreisen von Ulrich Jasper Seetzen. Seine in Italien gesammelten Kunstwerke und Altertümer sind heute Bestandteil der Sammlungen von Schloss Friedenstein.

Da sein älterer Bruder, Herzog August, 1822 ohne männliche Nachkommen starb, übernahm er als Friedrich IV. die Regierung des Herzogtums. Wegen seiner Krankheit hielt er sich lange zu Kuraufenthalten außerhalb seines Landes auf und regierte kaum selbst. Die Regierung führte für ihn sein Geheimrat Bernhard August von Lindenau. In seinen letzten Lebensjahren hatte er sein Sprachvermögen fast gänzlich verloren und war nur noch durch Gebärden in der Lage, sich seiner Umwelt zu vermitteln. Alle religiösen Fragen des Landes oblagen, wegen seines Glaubenswechsels, Friedrichs Ministerium. Während der Regierung Friedrichs IV. wurden die Hofhaltung, das Pageninstitut und die Leibgarde aufgelöst.

Nur drei Jahre nach seinem Regierungsantritt starb Herzog Friedrich IV. kinderlos. Weil ihm eine entsprechende Apanage verweigert worden war, hatte er sich nie vermählt. Mit ihm erlosch das Haus Sachsen-Gotha-Altenburg. Das Territorium fiel an die ernestinische Verwandtschaft und ging zum Teil im neu eingerichteten Herzogtum Sachsen-Coburg und Gotha auf.

Friedrich IV. wurde – wie zuvor schon sein Vater Ernst II. und sein Bruder August – auf der Insel im Großen Parkteich des Gothaer Schlossparks beigesetzt. Er ruht in der 1779 vom Architekten Carl Christoph Besser errichteten unterirdischen Gruft neben seinen im Kindesalter verstorbenen Brüdern Ludwig (gest. 1777) und Ernst (gest. 1779). Die einst die Gruft zierende Granitsäule mit einer Urne von carrarischem Marmor und Sockel von Serpentino antico des Bildhauers Friedrich Wilhelm Doell existiert seit Jahrzehnten nicht mehr, sodass der exakte Ort der Grablege Friedrichs IV. auf der Insel heute unbekannt ist.

Einzelnachweise

Literatur

Herzog (Sachsen-Gotha-Altenburg)
Wettiner (Linie Sachsen-Gotha-Altenburg, ab 1680)
Geboren 1774
Gestorben 1825
Mann

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