Oppression of women around the world Essays - Sex Crimes

Oppression of women around the world The Oppression of Women around the World and its Effects on Society Robyn Johnson PWSCC-Sociology-Professor Ballou The Oppression of Women around the World and its effects on Society "Of all the things that people do in the name of God, killing a girl because she doesnt bleed on her wedding night is among the most cruel." (WuDunn ,Kristoff, 2009, Half the Sky) Virginity is one of the most prized possessions a female can have. Its literally a matter of life and death in the Muslim world. Under Sharia law, If a woman is raped and there's loss of virginity the female can actually be charged with adultery and stoned to death. Take the case of 14 year old Hena Begum from Bangladesh India. One day her 14 year old cousin brutally raped her. After her rape, her rapist and his family beat her until she was unconscious. The local Sharia courts ruled that Hena committed adultery - for the crime of being a child raped by an adult. She was to receive 101 lashes. She could still not stand on her own when the Muslim clerics took her to the public square and proceeded to lash her. 80 lashes in she collapsed. Her parents were allowed to take her to the hospital, where she later died. Her father was ordered to pay $700.00 (US). Completely outrageous in such a poor country. The rapist had been charged with rape before and was forced to marry his victim. She coincidently was one of the people who initially helped beat Hena.(fortliberty.org,2014) It's hard for us in the USA to understand crimes and punishments such as these. To help us better understand we first need to really delve into the world of Sharia/Islamic Law. This is a system set forth 14+ centuries ago. Sharia law is considered the infallible law of God/Allah. Sharia law deals with many different things including politics, economics, crime, moral codes, and marital codes. One main thing to know is that women are completely oppressed and uneducated in regards to Sharia law. Amnesty International has found that 1 in 5 women will be a victim of rape in her lifetime. They also found that women are more likely to be beaten, raped, or killed by a current partner than by any other person. Studies estimate that 20-50 percent of women experience partner violence sometime in her lifetime. One hundred and two countries have no legal provisions against domestic violence. The UN estimates that five thousand women in the Middle East and Asia die a year as a result of honor kill ings. As of 2011 child marriages are common in the Middle East, accounting for 1 in 6 marriages in Egypt and 1 in 3 in Yemen. Rape is considered a crime in ALL Middle Eastern countries. Yet in some cases the rapist is forced to marry the rape victim. If she complains she can also be charged with rape and adultery and stoned to death.(Wikipedia, 2014) Many scholars claim that Sharia law encourages domestic violence against women, when a husband suspects Nushiz ( disobedience, disloyalty, rebellion, ill conduct) in his wife. Other scholars that is not consistent with modern perspective of the Quran. (wikipedia,2014) If we are to question why men and women in Muslim countries allow for such atrocities we will find that a huge factor is that Sharia law has done a very good job of keeping women uneducated. Men are also not very well educated. As long as you keep the masses uneducated they will follow like sheep. Then when you add in the raping, torture, stoning, cutting off of limbs. You can better grasp why Muslim's are at such a vulnerable position. Some think that Muslim men are afraid of women. Women create life. Women deal with all sorts of oppressive things and yet they march on. Sexual Slavery, Trafficking, and Prostitution The next severe form of oppression is sexual slavery, trafficking, and prostitution. In India, prostitution is legal yet there are many related activities that are not legal. Such as; soliciting in a public places, kerb crawling, owning or managing a brothel, pimping and pandering. There's an estimated 1.2 million children in prostitution in India. Some as young as 6. (UN.org,2013) Indian migrants

Attitude Crocodile tourism is one of the most famo Essays

Attitude Crocodile tourism is one of the most famo Essays Attitude Crocodile tourism is one of the most famous adventure tourism in the world that attracts a lot of tourist to visit Australia. The huge amount of tourists brings a lot of benefits to Australia, which includes huge amount of revenue for local government and tourist operators and great reputation in tourist industry under the economic downturn environment. However, the most famous tourism also result in some negative impacts breaking the ecological equilibrium and threatening local people ' s life. Therefore, it is hard for people to judge weather crocodile tourism is positive or not. Some individuals and environmental organizations considered that the crocodile tourism should be prohibited. On the other side, tourism operators will try their best to protect their profit as well. In order to retain the substantial income but reduce the negative impacts, negotiating and cooperation is the essential part that both sides need to attempt and put effort Beginning The most obvious and significant problem is the crocodiles becomes more aggressive and fearless to human, which is threatening people ' s life significantly. Due to the crocodile tourism, crocodiles are more familiar with boats and attracted by boats, further more, more humans were killed by crocodiles than before. According to the news, in 2014, saltwater crocodiles killed four Territorians in the Northern Territory, which is almost one-fifth of the total 21 people killed since 1974(NT News, 2014). The crocodile tourism has pushed not only natives closer to the dangerous but also backpackers. There were 31 victims on the list since 1990, all of them were killed by crocodiles in Australia, and there were many tragedies have not been recorded(C. Garry, 2011). The reasons why crocodiles attack human are multiple, which include territory defense, nest defense, self defense, food and by accident(C. Garry, 2011). To be more specific, crocodile is ec tothermic which means cold blood. They always become active when temperature increase, because they depends on solar energy. However, people are also prefer to travel during the warm period. Thus, season also could be summed as a factor(H. Melissa, 2015). Indeed, there are many factors result in crocodile attack, but , the core reason is the development of crocodile tourism and human ' s improper behaviors, over feeding and habitat lose changed crocodile ' s habits. Meanwhile, with the development of crocodile tourism, there are more chance for human to approach crocodile which means the situation could be worse in the future. Consider again The crocodile attack is not only threatening human ' s life, but also itself. Revenge kills are common in any place. According to the news report, in 2014, a African man killed crocodile after one month when his family was killed by crocodiles (H. Melissa, 2015). Further more, crocodile attack makes crocodile get a really bad reputation, which makes many innocent crocodile have been killed. In Australia, after a boy was missing, the authority in Australia issued a shoot-to-kill order on any crocodile bigger than two meters(H. Melissa, 2015). Thus, the bad reputation have put crocodile into a really dangerous edge. Some of the crocodiles are endangers species such as Philippine and Orinoco crocodile. However, most people do not have the knowledge of the crocodile, they will not consider them as Philippine and Orinoco, the only name of them is crocodile ' , the dangerous killer in the world. Therefore, the crocodile tourism has aggravated contradictions between human and crocodile to some extent. Definition of Stakeholders In order to define the stakeholders, it is better to look through the whole benefit chain in the crocodile tourism. On one hand, as an profitable industry, tourist industry is playing an essential role in Australia economy. According to the rep ort, during the period from 2013 to 2014, tourism has bring $102 billion income to Australia(Annual Report, 2013-14), which is a such big number. In the case, it also mentioned that in the decline trend of tourism industry, crocodile tourism still can remain a successfully number of visitors. So, we can find how much important is crocodile tourism for the tourism operators and the related industries such as hotel and catering industry, even many sole traders rely on the crocodile tourism. On the other hand, tourists also earn

Muslims who assimilate are better off than those who maintain their Essay

Muslims who assimilate are better off than those who maintain their traditions and culture - Essay Example The general belief among Islamic scholars is that "all men are not created equal, and democracy eventually lets the fortunate over-run the less fortunate. The idea of a caliphate is only now beginning to take hold in the Arab world. Europe won't come around until our example is there to follow "(Chu). Democracy in its present form is unacceptable to Islam. Islamic world believe that the efforts of Europeans to spread democracy across Arab world is an attempt to destroy Islamic culture and traditions. The agitations happening at present in Libya, Yemen, Syria, Bahrain, Oman, etc like Arab countries are perceived as the deliberate attempt by the Western World to topple Islamic administrations prevailing in those countries. It should be noted that these agitations were originated not because of the interferences of any other Western country, but because of the self-realization of people about their present pathetic conditions compared that in other countries. In fact the recent successf ul revolution in Egypt and the marginalization of people by the patriarchal administrations motivated the people in other Arab countries to fight against the fundamental Islamic administrations. â€Å"In the social realm, Shari'a is more restrictive than Western norms and lifestyles.

An entrepreneurial viewpoint Essay Example | Topics and Well Written Essays - 1500 words - 1

An entrepreneurial viewpoint - Essay Example Discerning the module has taught me that there are certain traits that any successful entrepreneur needs to be endowed with. A careful scrutiny of my own personality against these entrepreneurial traits has confirmed to me what I have and areas that I need to improve on to call myself a happy and successful entrepreneur. My entrepreneurial spirit is hemmed in by several personality traits that the module has helped me identify about myself and most immediately is confidence. Like Michael Bloomberg puts it: â€Å"Don’t be afraid to assert yourself, have the confidence in your capabilities, never letting the bastards drag you down.† I believe that confidence comes in handy for startup success. I have the confidence to take the risks, and to lead teams in creating a vision and to confidently sell the product to investors and customers (Straut, 2008). The other trait is optimism, Bill Gates has made fortunes as an entrepreneur because he looked at what was ahead of him with optimism; and that is how I always look at things that are ahead of me. I have learnt from the module that an entrepreneur should be optimistic about a product, their teams and their future. A study done by Baron (2004) found out that entrepreneurs perceive risks as being smaller than they are in actual sense, and also smaller than many people would ordinarily perceive them to be. It is with such optimism that I look at the future that though things may look obscured at the beginning, soon they will clear up, all it takes is to hang on and be positive. Passion has taught me that anyone can succeed in anything they set out to do, as long as their endeavor is anchored on unlimited and incredible enthusiasm. I am very passionate about everything I believe in and speaking about the importance of passion in entrepreneurship, Baron (2004, pp. 230) wrote, â€Å"Passion is a key cog in the wheel of determination, it is both contagious and palpable, many consumers, businesses and investors will be swayed by an entrepreneur who exhibits great passion.† Going with passion is self-efficacy. Nolan Bushnell was quoted saying that â€Å"many people have great ideas, but it’s only a few of them who decide to do something about those ideas now; a true entrepreneur is the one who is a doer, rather than a dreamer† (Choo, 2004). The last characteristic that wraps up my entrepreneurial spirit is persistence. I made a promise to myself that I was going to be successful in anything that I put my all into. There is no amount of money, or any amount of labor neither any number of hours would deter me from giving the very best that is in me. The module is clear that a successful entrepreneur is one who is will overcome and persevere despite the many hurdles, setbacks or failures (Osborne, 1995). Entrepreneurs are able to learn from their failures to make improvements on themselves, their team and their products. I have an entrepreneurial mentality as opposed to an employee mentality and so it would more fulfilling for me to work in an entrepreneurial organization as an entrepreneurial manager as opposed to working in a conveniently structured organization with convenient management approaches. This is inspired by the fact that I have a vision to chart and create my own path, as opposed to that employee mentality

Anabolic Steroids Essay Example | Topics and Well Written Essays - 2000 words

Anabolic Steroids - Essay Example The core of anabolic steroids touches two overlapping effects; anabolic and androgenic. Anabolic effect encourages cell growth, for masculization and maintenance of muscle; androgenic affect is used. Its use is nothing new. There are various medical and non-medical reasons of their use. The intake of these artificial hormones results in enhanced remodeling of bone growth and stimulation of bone marrow. Increased red blood cells, bigger appetite and greater than before protein synthesis. Mass lean muscle and increased weight in surprisingly short period of time may result by combining high dose of anabolic steroids with intensive exercise and high protein diet. When we talk about muscle and weight gain, it is just unavoidable to talk about "anabolic steroids." It is a drug, which makes the human body bigger and stronger, in a short span. It became very popular and found an instant way into athletes, body builders and weight lifter's life during 1940's.Most of the people use it to reduce body fat and get enlarged muscle size. It amazingly boosts the stamina and suppresses the stress hormone, cortisol. Due to this, anabolic steroids help the user to train more frequently. It also reduces the chances of muscle damage, injury and quicker recovery after the injury by acting as an anti-inflammatory. It is also used to keep the body fat level lower due to increased metabolic rate. The circumstances for muscle production become greater due to the increasing amount of nitrogen in the body. Most of the body builders and athletes make use of them for professional benefits. It gives strength and endurance to the user by increasing their ability to cop e with more physical pressure in today's cut throat competition of athletic world. Now getting down to the main point, anabolic steroid is used for medical purposes and still is a prescription based drug. With a passage of time its uses are becoming wider. Its main domain deals with hormonal problems in males, where there is not enough natural production of male hormone. "Hypogonadal states, delay of growth and puberty in boys, bone marrow failure, selected anemia, edema and late stages of breast cancer," Karch (1997) are the most common conventional medical uses include the use of anabolic steroid. In severe mal-nutrition and burnt cases, Anabolic steroids were taken, obviously with safety measures, by "post-operative patients to hasten recovery." Karch (1997). It ensures patient's quick recovery but due to lack of research, (ibid) "they have not proved efficacious for this appreciation." Latest investigational uses consist of endorsement of Anabolism in patients with Aids and alcoholic hepatitis, management of certain hyperlipemias and back-up for fibrinolyris. Administered dosage help the post-operative patients for quick recovery. It helps out people who lose muscle mass and face weight loss caused by Osteoporosis. It is also prescribed while treating some kinds of cancers. Anabolic steroids improve the symptoms without serious side effects in short cycles. Naturally, humans have various types of steroids present in their bodies. This commonly used drug is an artificial chemical compound, which intends to have a similar effect to those of a natural hormone. In males, who are unable to produce sufficient amount of testosterone, this seems to be the only

Chromium Induced Toxicity Research

Chromium Induced Toxicity Research Abstract In the present study, we hypothesize that cytotoxicity, genotoxicity and oxidative stress play a key role in chromium induced toxicity in SISS, SISK, IEE, IEK, IEG, SICH and ICG cell lines when exposed for 24 h. Acute toxicity tests were conducted on three fish species namely L. calcarifer, E. suratensis and C. catla by exposing them to different concentration (0, 10, 20, 30, 40 and 50 mg/L) of chromium for 96 h under static conditions and the LC50 was calculated. The percentage cell survival was assessed by multiple endpoints such as MTT, NR, AB and CB assays were performed in seven fish cell lines exposed to different concentrations of chromium and EC50 values of all the four endpoints was calculated. Linear correlations between each in vitro cytotoxicity assay and the in vivo mortality data were highly significant. Microscopic examination of cell morphology indicated cell shrinkage, cell detachment, vacuolations and cell swelling at highest concentration of chromium (50mg/L). The DNA damage and nuclear fragmentation were assessed by comet assay and Hoechst staining, in seven fish lines exposed to different concentrations of chromium. The result of antioxidant parameter obtained show significantly decreased catalase (CAT), superoxide dismutase (SOD), glutathione S-transferase (GSH) and Glutathione peroxidase (GPx), and increased level of lipid peroxidation (LPO) in all the cell lines after exposure to increasing chromium in a concentration-dependent manner. This results proves that fish cell lines could be used as an alternative to whole fish using cytotoxicity, genotoxicity and oxidative stress assessment after exposure to chromium. Keywords: Fish cell lines, Chromium, Cytotoxicity, Genotoxicity, Oxidative stress 1. Introduction Heavy metal pollution of water is a serious environmental problem facing the modern world. At global level heavy metals pollution is increasing in the environment due to increase in number of industries (Chidambaram et al. 2009). Industrial effluents are discharged into the sewage canals, rivers and irrigation water, causing major pollution and health hazards (Baddesha and Rao 1986). Many industrial wastewaters contain heavy metals like cadmium, lead, zinc, cobalt and chromium. The toxic heavy metals are mostly absorbed and get accumulated in various plant parts as free metals which may adversely affect the plant growth and metabolism (Barman and Lal 1994). Human beings and cattle are badly affected when these metals are incorporated into food chain as it causes bronchitis and cancer (Khasim et al. 1989; McGrath and Smith 1990; Nath et al. 2005). Among heavy metals, chromium plays a major role in polluting our aquatic environment system. In nature chromium occurs predominately in two valances Cr (III) and Cr (VI). Hexavalent chromium [Cr (VI)] predominates over the Cr (III) form in natural waters. Hexavalent chromium [Cr (VI)] particulates enter the aquatic medium through effluents discharged from leather tanning, textiles, chrome electroplating, metal finishing, dyeing and printing industries and several other industries. The Cr (VI) penetrates biological membranes easily and causes cellular damage by oxidative stress (Irwin et al. 1997; Begum et al. 2006), its unselective exposure may pose serious effect on aquatic communities including fish. Toxic effects of Cr(VI) on enzymological/biochemical (Al-Akel and Shamsi 1996; Vutukuru et al. 2007; Oner et al. 2008), hematological (Gautam and Gupta 1989; Al-Akel and Shamsi 1996), immunological (Prabakaran et al. 2007) parameters, endocrine toxicity (Mishra and Mohanty 2009) and genotoxicity (Chen et al. 2011) have been reported in many teleosts fishes. In environmental risk assessment, much of the toxicity test on fish has involved the use of lethality as the endpoint. On the other hand, in vivo bioassay is expensive and requires huge quantity of toxicant. The exposure time is only 24 h as opposed 96 h in bioassay, which could reduce the cost of labor, lab facilities and test time but more importantly allow decisions to be made more rapidly. Nevertheless, toxicity testing with fish is an essential part of environmental risk assessment procedures (Castano et al. 2003). For all these considerations, the development and use of in vitro assays that could measure early stages of toxicity in vertebrates represent an approach that could be very useful to monitoring environmental risk assessment (Walker 1999). Over the last four decades, cell and tissue culture methods have been refined and have now become an essential tool in environmental research. There are a lot of ethical, scientific and economical reasons that support the development of in vitro methods for use in ecotoxicology (Castano and Gomez-Lechon 2005; Bols et al. 2005; Schirmer, 2006; Fent 2007; Taju et al. 2012, 2013, 2014). The use of fish cell lines in environmental toxicology has been reviewed and positively assessed mainly with regards to cytotoxicity (Babich and Borenfreund 1991; Castano et al. 2003; Fent 2001). Cytotoxicity assessments can be readily employed to examine multiple endpoints, including measurements of cell death (apoptosis), cell viability, cellular morphology, cell metabolism, cell attachment/detachment, cell membrane permeability, proliferation, growth kinetics, genotoxicity and oxidative stress (Maracine and Segner 1998; Li and Zhang 2002; Shuilleabhain et al. 2004; Taju et al. 2014). In the present study, three fish species from three different aquatic environments, Lates calcarifer (Marine), Etroplus suratensis (Brackishwater) and Catla catla (freshwater) were selected as representatives of their respective environments to study their suitability for acute toxicity test to evaluate the potential risk of chromium (Cr). They are excellent food fishes with a good market demand in India, Malaysia, Bangladesh and Pakistan. Some attempts were made to study in vivo acute toxicity in Sea bass, Etroplus and Catla using various toxicants (Chezhian et al. 2010; Azmat and Javed 2011, 2012; Bhat et al. 2012; Taju et al. 2012, 2013). The seven fish cell lines namely SISK and SISS cell lines derived from L. calcarifer (Sahul Hameed et al. 2006; Parameshwaran et al. 2006b), SICH and ICG cell lines derived from C. catla (Ishaq Ahmed et al. 2009b; Taju et al. 2014), and IEE, IEK and IEG cell lines derived from E. suratensis (Sarath Babu et al. 2012) were used as in vitro assays t o evaluate the cytotoxicity, genotoxicity and oxidative stress exposed to chromium. The results of in vitro cytotoxicity were compared with the results of in vivo acute toxicity test using fish. The use of these cell lines for toxicity assessment of chromium instead of living fish is recommended. 2. Material and methods 2.1. Chemicals and reagents Tissue culture media and chemicals were obtained from GIBCO (Invitrogen Corporation, USA). Potassium dichromate (K2Cr2O7), EDTA, Trichloroacetic acid, DTNB [5,5-dithio-bis-(2-nitrobenzoic acid)], Thiobarbituric acid, Hydrogen peroxide, Nitro blue tetrazolium (NBT), Riboflavin, Hydroxylamine-HCl, Triton X-100, Ethidium bromide, Methanol, Acetic acid, Sodium chloride, Sodium hydroxide and Coomassie Blue was purchased from SRL chemicals, India. 2.2. Collection of experimental animals Lates calcarifer and Etroplus suratensis were collected from Central Institute of Brackishwater Aquaculture (CIBA), Chennai. Catla catla was collected from a local pond in Walajapet, Vellore District, Tamil Nadu, India. The experimental fishes were 2 3 g in body weight. Specimens were transported live in oxygen bags or buckets to the laboratory, acclimatized and maintained for 20-30 days in a salinity range of 5-10 ppt for E. suratensis, 20-25 ppt for L. calcarifer and in freshwater in the case of C. catla (23-28oC) under an ambient photoperiod in the laboratory for 10 days prior to experiments. The fish were fed with commercial pellet feed twice a day and starved for 24 h before and during the experiments. 2.3. In vivo fish acute toxicity test Fish acute toxicity tests were conducted by exposing E. suratensis, L. calcarifer and C. catla (N = 10 per aquarium) for 96 h to chromium under static conditions (OECD 203, 1992). Five different concentrations chromium i.e., 0, 10, 20, 30, 40 and 50 mg/L diluted with seawater (5 ppt) and freshwater while control with sea water and freshwater alone were tested to determine the LC50 (concentration at which 50% of the fish population dies). The aquaria had a working volume of 30 lit based on the body weight of fishes (1 g fish/L). Dead fishes were counted and removed immediately every day. All the experiments were conducted in triplicates. Mortalities were recorded following the guideline for fish acute toxicity OECD 203 (1992). 2.4. Fish Cell lines A total of seven cell lines established from different organs of L. calcarifer (SISS-seabass spleen, SISK-kidney), E. suratensis (IEE Etroplus eye, IEG gill, IEK kidney) and C. catla (SICH Catla heart, ICG gill) were tested for their sensitivities to chromium. These fish cell lines were propagated at 28oC in Leibovitzs L-15 medium (pH 7.0 -7.4) with 2mM L-glutamine, 10% foetal bovine serum (FBS), penicillin 100 IU/ml and streptomycin 100 ÂÂ µg/ml. The osmolarity ranged from 300 to 360 mOsm kg-1. These cells were sub-cultured every 2-3 days using standard procedure. Cells at exponential growth phase were harvested and used for in vitro cytotoxicity tests. 2.5. In vitro cytotoxicity assay using fish-derived cell lines SISS, SISK, IEE, IEK, IEG, SICH and ICG cells at exponential growth phase were collected and diluted to a concentration of 105 cells/ml in Leibovitzs L-15 medium with 10% FBS. After agitation, the cells were added to each well of 96-well tissue culture plates at the concentration of 2 x 104/well and incubated overnight at 28oC. After incubation, the medium was removed and the cells were re-fed with medium containing 0 (control), 10, 20, 30, 40 and 50 mg/L of chromium for 24 h EC50 analysis. Then four endpoints for cytotoxicity, i.e., MTT [3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide] assay, Neutral red (NR) uptake assay, Alamar blue assay (AB) and protein concentration for Coomassie blue (CB) assay were determined after 24 h exposure as described by Borenfreund et al. (1988), Borenfreund and Puerner (1985), Taju et al. (2012) and Shopsis and Eng (1985), respectively. 2.5.1. Cell morphology SISS, SISSK, IEE, IEK, IEG, SICH and ICG cells were plated into a 24 well tissue culture plate at a density of 2ÃÆ'-105 cells (in 1 mL growth medium). After overnight growth, supernatants from the culture plates were removed and fresh aliquots of growth medium containing various concentrations of the chromium (0, 10, 20, 30, 40 and 50 mg/L) were exposed for 24 h. Upon incubation, cells were washed with phosphate-buffered saline (PBS, pH 7.4) and the morphological changes were observed under an inverted phase-contrast microscope (Carl Zeiss, Germany) at 100ÃÆ'- magnification. 2.6. Assessment of in vitro genotoxicity using fish-derived cell lines 2.6.1. Comet assay The Single Cell Gel Electrophoresis (comet assay) was performed on SISS, SISK, IEE, IEK, IEG, SICH and ICG cell lines according to the method of Singh et al. (1988) with slight modifications in accordance with the protocols of Taju et al. (2014). 5 x 104 cells on 500 ÃŽÂ ¼L of complete culture medium were seeded per well in a 24-well-plate. After 24 h incubation, cells were exposed to chromium using the following concentrations: 0 (control), 10, 20, 30, 40, 50 and 60 mg/L. At the end of the exposure period, cells were collected through trypsinization, followed by centrifugation at 1000 rpm for two minutes to obtain the pellet and avoid cell loss. After the centrifugations, the supernatant was discarded and the pellet resuspended in 100 ÃŽÂ ¼L of 0.9% agarose in milliQ water (low-melting point agarose, Sigma Aldrich chemicals, USA). The suspensions of cells in agarose were then applied dropwise to microscope slides containing an agarose layer (agarose electrophoresis grade, prep ared with a 1% concentration in milliQ water), and kept in a freezer for 10 min. The cells were lysed in freshly made lysing solution (2.5 M NaCl, 100 mM EDTA, 10 mM Tris-HCl, 10% DMSO, 1% Triton X-100, pH 10), for 1 h at 4 ÂÂ °C. After rinsing with redistilled water, the slides were placed on the horizontal gel box, covered with the cold alkaline buffer (0.3 M NaOH, 1 mM EDTA, pH >13) and left for 20 min. Electrophoresis was run in the same buffer at 25 V (0.83 V/cm) at 300 mA for 20 min at 4 ÂÂ °C. After electrophoresis the slides were neutralized in a cold neutralization buffer (0.4 M Tris-HCl, pH 7.5), for 2 to 5 min, fixed in methanol:acetic acid (3:1) for 5 min and stored in the dark at room temperature. Prior to examination, the slides were rehydrated and stained with 10 ÂÂ µg/mL ethidium bromide and examined using a Zeiss Axioplan epifluorescence microscope (Carl Zeiss, Germany). A positive control (5 ÂÂ µM H2O2) was also included in every batch of sample s. This strategy was chosen to compare the variation in the distance of migration. The positive control was not included in evaluation. Slides were examined at 100x magnifications using a fluorescence microscope. For each experimental condition 100 randomly chosen cells from two duplicate slides were examined (50 from each slide). In all 100 comets were scored visually according to the relative intensity of the fluorescence in the tail length. The extent of DNA migration was determined as a percentage of DNA in the tail (% tDNA) using an image analysis system comet 5, Kinetic Imaging Ltd. 2.6.2. Assessment Nuclear fragmentation by Hoechst 33258 Nuclear fragmentation of SISS, SISK, IEE, IEK, IEG, SICH and ICG cell lines was analyzed with Hoechst 33258. The cells were seeded in 12-well cell culture plates and incubated overnight. Then the cells were treated with different concentrations of chromium (0, 10, 20, 30, 40 and 50 mg/L). Cells were fixed in 4% paraformaldehyde in PBS for 30 min, washed with PBS, and stained with 1 ÃŽÂ ¼g/mL Hoechst 33258 in PBS for 30 min. Stained cells were washed twice with PBS. The changes in nuclei were observed with a fluorescent microscope through a UV filter. 2.7. Preparation of cell extract and Biochemical estimations The SISS, SISK, IEE, IEK, IEG, SICH and ICG cell lines were exposed to different concentrations of chromium (0, 10, 20, 30, 40, 50 and 60 mg/L) on 25 cm2 flasks for 24 h. After 24 h they were trypsinized and pelleted by centrifugation at 500ÃÆ'-g for 5 min. The cell pellet was washed with PBS (0.1M, pH7.4), resuspended in 500 ÂÂ µl chilled homogenizing buffer (250mM sucrose, 12mM Tris-HCl, 0.1mM DTT, pH 7.4) and lysed using Dounce homogenizer. The lysate was centrifuged (8000ÃÆ'-g, 10 min, 4 ÂÂ °C) and the supernatant (cell extract) was used for various biochemical assays. Protein concentration in the cell extract was estimated by the method of Lowry et al. (1951). The enzymatic antioxidant superoxide dismutase (SOD) activities were determined by following the procedures described by Kono (1978). Catalase (CAT) activity was determined by following the method described by Aebi (1974). The level of non-enzymatic antioxidant reduced glutathione (GSH) was estimated following the procedures described Saldak and Lindsay (1968). The activity of glutathione peroxidase (GPx) was assayed by the method of Flohe and Gunzler, (1984). The level of lipid peroxidation (LPO) was measured according to the method described Beuge and Aust (1978) based on the reaction with thiobarbituric acid. The results were recorded as ÂÂ µmol of TBA reactive substances/mg protein. The enzymatic and non-enzymatic parameters was expressed as ÂÂ µmol/mg protein. 2.8. Data analysis Experiments were performed in triplicate with eight replicates for each exposure concentration. Absolute values of each assay were transformed to control percentages. The results of LC50 and EC50 values were expressed as dilution in (mg/L) of the sample calculated using computerized (EPA, 2000) software. The individual data points of the concentration response cytotoxicity graph were presented as the arithmetic mean percent inhibition relative to the control standard error (SE). Cell viability and the concentration were fitted Scatter plots with the regressive equation (a linear regression model). The strength of the r2 value was used to determine whether a linear or quadratic relationship was assumed. Analysis of variance was used to determine whether groups of variables differed from each other (SPSS, Version 16). 3. Results The cumulative percentage mortality in L. calcarifer, E. suratensis and C. catla exposed to different concentrations of chromium was determined at 96 h and the results are presented in Fig 1. The toxic effect of chromium on the survival of fish was found to be concentration and time dependent. The chromium at the concentration of 50 mg/L caused 100%, 96.66% and 90% mortality, respectively, in L. calcarifer, E. suratensis and C. catla, whereas lower concentration of chromium at 10 mg/L caused 26.66%, 16.66% and 20% mortality of L. calcarifer, E. suratensis and C. catla respectively. No mortality was recorded in the control fish even after 96 h exposure. The LC50 values corresponding to 24, 48, 72 and 96 h of exposure of chromium were determined and results are presented in Table 1. Five different concentrations which ranged from 10 to 50 mg/L of chromium were used to carry out the in vitro toxicity assay in SISS, SISK, IEE, IEK, IEG, SICH and ICG cell lines using four cytotoxicity end points (MTT, NR, AB and CB assays) and the results are shown in Fig.2 A-D. The cytotoxicity of chromium to SISS, SISK, IEE, IEK, IEG, SICH and ICG cell lines was found to be similar in all the toxic endpoints employed. The lowest concentration of chromium tested (10 mg/L) was found to toxic in all the cell lines particularly SICH and IEK cell lines. The progressive increase in the concentration of chromium led to increase in toxicity when compared to control cells. The MTT, NR, AB and CB cytotoxicity endpoint assays revealed that a 24-h exposure of all the cell lines to different concentrations of chromium produced a dose-dependent reduction in the fraction of viability. The EC50 values and 95% confidence limit values obtained for chromium are summarized in Table 2. Correlations a mong the endpoints employed in the SISS, SISK, IEE, IEK, IEG, SICH and ICG cell lines to study cytotoxicity of chromium have been determined. A general tendency in the sensitivity among the four endpoints could be observed and statistical analysis revealed good correlation with R2 = 0.889-0.927 for all combinations between endpoints (Data not shown). The in vivo values of L. calcarifer vs. in vitro data of its two cell lines exposed to chromium were highly significant p2=0.956 (L. calcarifer vs. SISS) and 0.962 (L. calcarifer vs. SISK); R2=0.973 and 0.993; R2=0.980 and 0.975; R2=0.992 and 0.977 for MTT (Fig 3A), NR (Fig 3B), AB(Fig 3C) and CB (Fig 3D), respectively. The in vivo values of E. suratensis were compared with in vitro values of its three cell lines (IEE, IEG and IEK) exposed to chromium and were found to be highly significant p2=0.985 (E. suratensis vs. IEE), 0.987 (E. suratensis vs. IEK) and 0.968 (E. suratensis vs. IEG); R2=0.980, 0.936 and 0.956; R2=0.961, 0.955 and 0.904 and R2=0.955, 0.939 and 0.974 for MTT (Fig 3E), NR (Fig 3F), AB(Fig 3G) and CB (Fig 3H), respectively. Linear correlations between each in vitro vs. in vivo (C. catla)values of chromium were highly significant p2=0.991 (C. catla vs, SICH) and 0.993 (C. catla vs, ICG); R2=0.982 and 0.983; 0.974 and 0.990 and 0.987 and 0.984 for MTT (Fig 3I), NR (Fig 3J), AB(Fig 3K) and CB (Fig 3L), respectively. The prominent morphological changes of the cells exposed to high concentrations of chromium were observed. The changes observed include cell shrinkage, cell detachment, vacuolations and cell swelling in SISS (Fig 4H), SISK (Fig 4I), IEE (Fig 4J), IEK (Fig 4L), IEG (Fig 4L), SICH (Fig 4M) and ICG (Fig 4N) cell lines. In controls, no morphological alterations were observed in the SISS (Fig 4A), SISK (Fig 4B), IEE (Fig 4C), IEK (Fig 4D), IEG (Fig 4E), SICH (Fig 4F) and ICG (Fig 4G) cell lines. The percentage of DNA damage and the cumulative tail length from 100 cells per sample were measured in SISS, SISK, IEE, IEK, IEG, SICH and ICG cells exposed to different concentrations of chromium (0, 10, 20, 30, 40 and 50 mg/L) and the results are shown in Fig. 5. The length of tail DNA in SISS, SISK, IEE, IEK, IEG, SICH and ICG cells exposed to 10 mg/L of chromium was estimated to be about 1.7%, 2.0%, 1.3%, 1.5%, 2.1%, 1.4% and 1.5%, respectively at a 24-h exposure, and chromium at the concentration of 50 mg/L caused 8.9%, 11.0%, 9.4%, 8.8%, 11.1%, 6.4% and 7.2% of tail DNA migration in SISS, SISK, IEE, IEK, IEG, SICH and ICG cells, respectively (Fig. 5). Comet results of chromium exposed SISS, SISK, IEE, IEK, IEG, SICH and ICG cells showed a dose dependent increase in tail DNA (%) compared to the control cells, which gave the extent of DNA damage. The SISS, SISK, IEE, IEK, IEG, SICH and ICG cells were exposed to chromium for 24 h at different concentrations (0, 10, 20, 30, 40 and 50 mg/L) and the results are shown in Fig. 6A-N. Apoptotic cells were identified by Hoechst staining of condensation and fragmentation of the nuclei as shown in SISS cells (Fig. 6H), SISK cells (Fig. 6I), IEE cells (Fig. 6J), IEK cells (Fig. 6K), IEG cells (Fig. 6L), SICH cells (Fig. 6M) and ICG cells (Fig. 6N) at higher concentration i.e. 50 mg/L of chromium exposed for 24 h, while no nuclear changes were observed in control cells are shown in SISS cells (Fig. 6A), SISK cells (Fig. 6B), IEE cells (Fig. 6C), IEK cells (Fig. 6D), IEG cells (Fig. 6E), SICH cells (Fig. 6F)and ICG cells (Fig. 6G). The level of antioxidant parameters such as SOD, CAT, GPx, GSH and LPO was measured in SISS, SISK, IEE, IEK, IEG, SICH and ICG cells exposed to different concentrations of chromium and the results were presented in Fig 7A-E. Regarding oxidative stress biomarkers, no significant change was observed in SOD, CAT, GSH and LPO levels in the SISS, SISK, IEE, IEK, IEG, SICH and ICG cells exposed to lower concentrations i.e. 10 mg/L of chromium when compared to the control cells. However, when these cell lines were exposed to 50 mg/L of chromium, the activity of SOD (~2.1, ~2.3, ~1.5, ~1.3, ~2.3, ~1.2 and ~2.2 fold in SISS, SISK, IEE, IEK, IEG, SICH and ICG cells respectively in Fig 7A), CAT (~5.2, ~6.8, ~5.3, ~7.4, ~6.4, ~5.2 and ~4.6 fold in SISS, SISK, IEE, IEK, IEG, SICH and ICG cells respectively Fig 7B) and level GSH (~1.6, ~1.5, ~1.3, ~1.6, ~1.5, ~1.8 and ~1.3 fold in SISS, SISK, IEE, IEK, IEG, SICH and ICG cells respectively Fig 7C) and GPx (~1.2, ~1.1, ~1.0, ~1.2, ~1.1, ~0.9 and ~1. 3 fold in SISS, SISK, IEE, IEK, IEG, SICH and ICG cells respectively Fig 7D) decreased was found to be significantly (*P 4. Discussion Heavy metals constitute a main group of aquatic pollutants due to their bioacuumulative and non-biodegradable properties (Velma and Tchounwou 2010). Their excessive contamination of aquatic ecosystems has evoked major environmental and health concerns worldwide (Vutukuru et al. 2007). Chromium is the sixth most abundant heavy metal in the earth crust (U.S. EPA 1984). Fish and Fish cell lines constitute an excellent model to understand the mechanistic aspects of metal toxicity (Taju et al. 2014). In this study, we have examined the in vivo toxicity in three fish species in different environment i.e. L. calcarifer (Marine water), E. suratensis (brackish water) and C. catla (Fresh water), and in vitro cytotoxicity, oxidative stress and genotoxicity of the three same fish cell lines, SISS, SISK (Seabass spleen and kidney cell lines), IEE, IEK, IEG (Etroplus eye, kidney and gill cell lines), SICH and ICG (Catla heart and gill cell lines) an exposure to chromium. The results of this study clearly show that the fish cell lines experienced oxidative stress by modulating the antioxidant enzyme, exhibited DNA damage, nuclear fragmentation and microscopic morphological changes in the SISS, SISK, IEE, IEK, IEG, SICH and ICG cells. The LC50 values of chromium were determined as 30.22, 33.83 and 30.64 mg/L respectively in L. calcarifer, E. suratensis and C. catla, respectively at 96 h of exposure in this study. Recently, Mishra and Mohanty (2009) reported the LC50 values of chromium on Channa punctatus at 96 h of exposure as 41.75 mg/L. The LC50 values observed by Mishra and Mohanty (2009) were found to be higher when compared to L. calcarifer, E. suratensis and C. catla and this indicates that the L. calcarifer, E. suratensis and C. catla were found to more sensitive to chromium. Seven fish cell lines derived from L. calcarifer (SISS SISK), E. suratensis (IEE, IEK and IEG) and C. catla (SICH and ICG) were applied to evaluate the cytotoxicity of chromium using MTT, AB, NR and cell protein (CB) assays. The results of in vitro assays were compared with the results of in vivo test to determine the suitability of these fish cell lines for toxicological studies to replace the use of whole fish. The evaluation of cytotoxicity of chemical substances using animal cells has been carried out by many workers (Ekwall 1980a, 1983; Metcalfe 1971; Muir 1983a, 1983b; Paganuzzi et al. 1981; Benoit et al. 1987). Four commonly used endpoint assays (MTT, NR AB and cell protein assay CB) were employed in the present study using SISS, SISK, IEE, IEK, IEG, SICH and ICG cell lines of E. suratensi, C. catla and L. calcarifer to evaluate the in vitro cytotoxicity of chromium. The main observation was that the cytotoxicity was closely associated in all the seven cell lines independent of the toxic endpoints employed. This not only supports the observations of Ekwall (1995) and Li and Zhang (2002) that most cell lines have a similar results to toxicants when toxicity is measured by different endpoints, corresponding to inhibition or destruction of basal functions and structures, and also suggests that endpoints employed in the present study can also be used to predict acute cytotoxicity. Tan et al. (2008) have used six fish cell lines to study the toxicity of four heavy metals: cadmium, chromium, zinc, and copper by using two cytotoxicity endpoints MTT and CB assays. The results revealed that carp epithelioma cells are least tolerant to chromium. The NR uptake assay is a useful method for comparing the relative acute cytotoxicity of metals in vitro with metal and chemicals toxicity studies in whole fish in vivo (Brandao et al. 1992; Ryan and Hightower 1994; Taju et al. 2013). In the present study, we employed that SISS, SISK, IEE, IEK, IEG, SICH and ICG cell lines for cytotoxicity assessment of chromium by using four endpoints. Our results show that there is no significant difference between all the four endpoints. Segner (1994) reported that the relationship of the in vitro cytotoxicity values to in vivo fish toxicity data is less satisfying and that this might be due to the inconsistency of the in vivo values. As observed in the present study, a positive relationship of acute lethal potency in fish with in vitro cytotoxicity has been found by Fry et al. (1990). Castano et al. (1996) found good correlations between in vivo and in vitro for each endpoint and for the cytotoxicity index and suggested the applicability of the RTG-2 cell line as an alternative protocol to estimate the acute toxicity of chemicals on fish without using live animals. The correlation of in vitro cytotoxicity of metals with in vivo toxicity data was evaluated by comparing the 24 h NR50 results of R1 cells to 96 h LC50 data of different fish species. The rvalues (R1 cell line) were 0.64 for the relation between LC50, data of golden ide and bluegill sunfish, 0.58 for golden ide and rainbow trout in soft water, and 0.68 for golden ide and rainbow trout in hard water (Segner et al. 1994). In the present study, in vitro cytotoxicity of chromium with in vivo results was evaluated by comparing the 24 h MTT, NR, AB and CB data of seven Indian fish cell lines to 96 h data of three fish species (L. calcarifer, E. suratensis and C. catla). A good correlation was found between in vitro of seven fish cell lines compared with in vivo values of whole fish exposed to chromium for 24 h and 96 h respectively, with r=0.902 to 0.99. The results revealed that the four endpointsvalues were closely correlated to whole fish in vivo values and that the linear correlation b etween each in vitro parameter and the in vivo data were found to be highly significant. The results of in vitro assays using SISS, SISK, IEE, IEK, IEG, SICH and ICG cell lines of E. suratensis, C. catla and L. calcarifer were correlated with those obtained from in vivo assay using the same species of fish (L. calcarifer, E. suratensis and C. catla). Based on the results of the present study we recommend the use of these seven cell lines instead of living fish for toxicity assessment of metal salts and environmental contaminants. The present study showed that chromium induced genotoxicity in SISS, SISK, IEE, IEK, IEG, SICH and ICG cell lines by comet assay. DNA damage was observed in SISS, SISK, IEE, IEK, IEG, SICH and ICG cell lines exposed to chromium in a concentration dependent manner. The DNA damage at higher test concentrations in SISS, SISK, IEE, IEK, IEG, SICH and ICG cell lines could be due to the elevated levels of tail DNA in all cell lines compared to their controls cells. Induction of ROS under metallic stress could attack the DNA and damage its integrity. Our present results are similar to the previous reports (Iqbal Ahmad et al. 2006; Velma and Tchounwou 2010; 2013) DNA damage in gill and kidney of Anguilla anguilla L. exposed to chromium with or without pre-exposure to ÃŽÂ ²-naphthoflavone. In another study, medaka fin cell lines exposed to Cr (VI) to examine the genotoxic potentials, have observed DNA double strand breaks a

Analysis of The Revolt of Mother Essay -- Analysis of The Revolt of Mo

Analysis of The Revolt of Mother â€Å"The Revolt of ‘Mother’† by Mary Wilkins Freeman, was a story of a woman who lived in New England around or before the author’s time. The mother, Sarah Penn, was kept out of the families decisions by the father, Adoniram Penn, until one event that lead to her taking drastic actions while her husband was gone. There are many religious symbols and actions taken by â€Å"Mother† within the story. Through the story Sarah moved from a feeling of servitude to her husband, to a feeling that she was in servitude to the Lords will and this led her, in the end, to hold power over her husband. The religious overtones start with the title of the story, â€Å"The Revolt of ‘Mother.’† The name ‘Mother’ in many stories is used to relate to a divine or spiritual woman. It could be a direct reference to Mother Mary, but in the context of this story it is just meant to signify her clarity with what the Lord wants her to do. The word ‘Revolt’ also has religious significance when related with that use of ‘Mother’. The revolt that ‘Mother’ takes is a religious one because it is going against her husband and town’s beliefs, which are both the same. This becomes clear in the part of the story when the minister comes to talk to Sarah about what she has done. When the minister came to see Sarah on Friday after she had moved into the barn she was described as having a â€Å"saintly expression of her face†(529). With that, and the fact that she acts so rude to him, especially since he’s a minister, shows that she does believe she is right under the Lords will and he is not. The author also implies this by the name she gives to the minister, Mr. Hersey. His name sounds just like the word heresy and is spelled very similar. This is another indication that, in fact, the minister is going against the Lords own will and Sarah is not. The narrator has also described the minister as being â€Å" a sickly man† and that, â€Å"he had scourge himself up to some of his pastoral duties as relentlessly as a Catholic ascetic†(530). This seems harsh at first but it is just conveying the ignorance of the minister. In the times of this story the town would have to be Protestant or maybe even Puritan, but definitely not Catholic because it is set in early New England times and for the simple fact that he is a minister with a wife. The fact that he is referred to as a â€Å"Cathol... ...other was able to start the revolt. By the end of the story Adoniram is not in power. This aspect is shown through how Father lacked the power to even remove his jacket, even though he is described as having a â€Å"sturdily healthy†(531) frame. He is also not referred to as father, but as an old man. Because old people are usually represented as being weaker and more needing of help, Father takes on the position of lesser power in the family. So because Sarah moved from Adoniram’s servitude to the Lords, father falls to a lesser power. It is in this position that he gives into his wife and builds windows and partitions as she asks. In the end Sarah has moved from servitude to a position of power in the family. And Adoniram has fallen from his position of not listening to what Sarah wants to one of submission to her needs. But there hasn’t been sufficient foundation laid by Sarah to foreshadow that it will stay this way forever. First because the majority of the revolt took place when Adoniram was absent, so it was easier for her to make this transition. But mainly because as the name implies this was only a revolt. Just because you won a revolt doesn’t mean you’ve won the war.