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In 2010 India launched the National Mission for Empowerment of Women (NMEW). Its mandate includes facilitating the co-ordination of all programmes connected to women's welfare and their socio-economic development across all Ministries and Departments. In addition, a National Resource Centre for Women has been set up and acts as a central repository for available information and research on all gender related issues. India ratified the Convention on the Elimination of All Forms of Discrimination against Women (CEDAW) in 1993.
India is a pluralistic society and its constitution allows personal laws. Personal laws vary according to religion, with India's plural legal system accommodating a combination of civil, religious and customary laws. The 1855 Hindu Marriage Act and the 1956 Hindu Succession Act govern the Hindu population. The 1937 Muslim Personal Law Sharia Application Act and the 1986 Muslim Women's Protection of Rights on Divorce Act govern the Muslim population. Christians and Parsis are governed by the Christian Marriage Act and the Parsi Marriage and Divorce Act.
Under civil law, the minimum age for marriage is 18 years for women and 21 years for men.[1] However, it is also governed in practice by customary law, which can provide for a lower age. For example, the Muslim Personal Law[2] (though not codified) gives Muslims the authority to determine when marriage is acceptable, which if typically understood to be the age of puberty.[3]Notwithstanding the legal marriage age, India continues to account for the highest number of early marriages in the world,[4] despite legal and policy commitments to eliminate the practice. According to United Nations data from 2006, 27.6% of girls between 15 and 19 years of age are married, divorced or widowed.[5] India's most recent national health and demographic survey found that 46% of all marriages in India are early marriages;[6] with rates varying from region from 11% to 60%.[7]
India pledged to eliminate early marriage by 2010 in its 2005 National Plan of Action for the Girl Child and adopted the Prohibition of Child Marriage Act (PCMA) in 2006, which establishes penalties for the marriage of girls below 18 and boys below 21 and renders such marriages voidable.[8]However, NGOs working in the area of early marriage note that implementation of the PCMA has been weak.[9]For example, the PCMA makes underage marriages voidable, (not automatically void), only by judicial authority; which may serve as a barrier for girls who lack the autonomy to access and pay for legal services.[10] Further, passage of the PCMA does not clarify whether it supersedes personal laws, which has led to ambiguity concerning whether the minimum ages of marriage and the status of early marriages as voidable should be universally applied, or if the ages of marriage and legal statuses of early marriage established under personal laws should prevail.[11]
Indian law requires the consent of both parties for a valid marriage to take place, however public opinion suggests that women may not always exercise this right in practice. Only 26% of those polled in 2007 Pew survey believed that a woman should be free to choose her spouse; 24 percent believed that a woman's family should choose for her, and 49% responded that they should share the responsibility.[12]
Legally, both spouses have equal rights within marriage. However in practice, fathers are considered to be the 'natural' head of the family and to exercise parental authority.[13]In cases of divorce, courts are instructed to take decisions regarding child custody in the best interests of the child. However, the 2005 CEDAW report notes a trend whereby in most cases, Family Courts award custody of small children and of girls to mothers, while fathers are more often awarded custody of boys and of older children.[14]
Rights to divorce differ for those who chose to marry under religious law. While Muslim men have the right repudiate (i.e. divorce unilaterally) their wives[15]; Hindu divorce may be equally predicated on wrongdoing (such as adultery)[16]; and Christian marriages can be dissolved by a man for any reason, while Christian woman can only initiate divorce on the grounds of adultery.[17] The civil law provides that women are entitled to maintenance as part of divorce. However, the Muslim Women (Protection of Rights on Divorce) Act 1986 restricts maintenance to Muslim women, by requiring the husband to give 'a reasonable and fair provision, and maintenance' to his ex-wife only within the Iddat period (before she remarries). [18]
The Hindu Succession Act of 2005 granted Hindu, Sikh, Buddhist and Jain women equal inheritance rights to ancestral and jointly owned property, although enforcement of this law is weak.[19] In 2011 the government reported that the section of the law that denied rights of a widow to inherit her husband's property upon her remarriage has been repealed.[20] In practice, many women, particularly in northern India, are still deprived of inheritance. A recent survey by UN Women conducted in two states found that only 22% of families surveyed knew about the 2005 amendment that gave women equal inheritance rights.[21] However, even if women are aware of this amendment, they tend to give this right away, for example to their brothers, to avoid any family tensions.[22] In addition, the Muslim population of India may follow inheritance guidelines set out in Sharia law which discriminate against women and girls; daughters, for example, inherit half as much as sons.[23]
The practice of dowry is still in use in many areas, notwithstanding the Dowry Prohibition Act of 1961, amended in 1986, which prohibits dowry and penalises the giver, taker and the persons demanding and encouraging it.[24]
Polygamy is legal for the Muslim population, but is illegal for all other groups.[25]The 2005-2006 Demographic Health Survey (DHS) found that 1.7% of married women aged 15-49 were in a marriage with one or more co-wives.[26]
Adultery is a crime under India's 1860 Penal Code, although it treats adulterous acts by men and women differently. Section 497 criminalizes adultery by a man with the wife of another man 'without this consent'.
[1] CEDAW (2005), p.93 [2] The Muslim Personal Law (Shariat) Application Act, No. 26 of 1937, India Code (1937) [3] CRR (2013), footnote 108. [4] WHO (2013) [5] UN (2012) [6] Government of India (2006), p.166 [7] UNFPA (2012), p.29 [8] Government of India, Ministry of Human Resource Development, Department of Women and Child Development, National Plan of Action For Children 2005, 18 (2005); Prohibition of Child Marriage Act, No. 6 of 2007, arts. 2(a), 3. I [9] CRR (2013), p.6 [10] Prohibition of Child Marriage Act, Art. 3 [11] Prohibition of Child Marriage Act, Arts. 2(a), 3 [12] Pew Research Center (2007), Question Q.44.[13] CEDAW (2005), p.45 [14] CEDAW (2005), p. 45 [15] CEDAW (2005), p.4 [16] The Hindu Marriage Act of 1955, Section 13 [17] UNDP (2010), p.127 [18] Muslim Women (Protection of Rights on Divorce) Act 1986, Art. 3 (1)a [19] Oxfam (2013), p.1 [20] CEDAW (2012), p.5 [21] UN Women (2012), p.5 [22] Basu, Srimati (2001) [23] The Muslim Personal Law (Shariat) Application Act, 1937, Art. 2 [24] FAO (n.d) [25] CEDAW (2005), p. 93 [26] Measure DHS Statcompiler (n.d.)
The Protection of Women from Domestic Violence Act 2005 prohibits domestic violence, and specialized court or procedure for cases of domestic violence.[27] Regardless of marital status or the application of different communal family laws, women can secure protective orders and file cases against defendants in the same residence.[28] 2013 amendments to the Penal Code also strengthened prohibitions against certain forms of domestic violence. For example, the punishment for an acid attack that causes harm to the victim will be a minimum term of 10 years' imprisonment, extendable to a life term, while conviction on voluntarily throwing or attempting to throw acid with the intention of causing damage will incur a penalty of five to seven years.[29] In practice however, the implementation of the law on domestic violence remains weak, due to lack of capacity and resources of police and other agencies, and widespread corruption in law enforcement.[30]
Reports of spousal violence have increased in recent years, although this may reflect increasing awareness of legal avenues for redress.[31] In 2012 the NCRB recorded 106,527 cases of 'cruelty by husbands or relatives', only 15 percent of which resulted in convictions.[32] In the 2006 Demographic and Health Survey, nearly 40% of married women reported having ever experienced physical, sexual, or emotional violence at the hands of their husband or partner, while 24% reported such violence occurred in the previous year.[33] Attempts to combat the problem are frustrated by the fact that, a majority of married people in India view domestic violence as acceptable under certain circumstances. When presented with a choice of seven reasons why a man may be justified in beating his wife, 54% of women and 51% of men agreed with at least one of the reasons, the most common of which was arguing with her husband (about 38%).[34]
In 2013, the Ministry of Health and Family Welfare has promulgated Guidelines and Protocols for dealing with Medico-legal treatment of survivors and victims of sexual violence. Sensitivity, timely intervention and coordination with the police are the defining principles that back these efforts.[35] Also, Lawyers Collective has released in 2014 a publication entitled "Locating the Survivor in the Indian Criminal Justice System: Decoding the Law" to guide and sensitize all stakeholders on legal issues involved in such cases, and to better understand the nuances of the criminal justice system. [36][37] In addition, the National Legal Services Authority (NALSA)[38]hold sessions for sensitising stakeholders to deal with cases of violence against women, and there is a toll-free 24x7 Women helpline (1091) available for reporting cases of violence against women[39]. There is also an Umbrella Scheme for Protection and Empowerment of Women
In response to the public outcry following the death of a 23 year-old woman gang raped in New Delhi, the Government of India passed specific 'anti-rape' provisions into the criminal law, via the Criminal Law (Amendment) Act 2013. Under the Amendment, the Penal Code now provides for a sentence of seven years to life imprisonment and a fine for rape; ten years to life if the crime is committed by a person in a position of trust and authority such as police officers, a relative, guardian, or teacher[40]; and provides for capital punishment if the act causes the victim's death or leaves her in a permanent vegetative state.[41] Gang rape is made punishable by a prison term of from 20 years to life and a fine.[42] The Act also seeks to strengthen access to justice by making public servants who improperly conduct investigations in gender-based violence crimes, subject to a punishment of from six months to two years of imprisonment and a fine[43]; and hospital staff who fail to treat victims punishable by a prison term of up to one year and/or a fine.[44]
The 2012 Protection of Children from Sexual Offences (POCSO) Act defines and describes sexual offences including sexual assault and provides for strong punitive measures against offenders. It offers protection for all children including girls who are victims of offences defined under the Act and lays down child friendly procedures for investigation and handling of such cases.[45]
Although these provisions are far reaching and improve access to justice for rape victims, one UN Expert has stated that they do not go far enough to address the systemic inequality that fuels sexual violence.[46] In addition the amendments still fail to criminalize rape within marriage, which is not covered by the Indian Penal Code unless it involves a girl below the age of 15.[47] There is also a question of whether the courts are resourced and able to implement the new provisions; given findings by the US Department of State that legal avenues for rape victims are inadequate, and overburdened.[48] In addition, families frequently discourage reporting of rape: their attitudes tend to be about 'family honour' rather than the plight of the rape victim.[49]
According to figures from the federal Home Ministry's National Crime Records Bureau (NCRB) 24,923 cases of rape in 2012 were reported, up from 22172 cases in 2010 (a 12.4% increase),[50]with a 24% conviction rate. Yet, it is difficult to establish whether incidences of rape have increased or if there is more reporting. [51] The police also reported 8,233 cases of dowry death in 2012, and 38,262 cases of kidnapping and abduction.[52] Notwithstanding these high figures, reported cases still represent only a fraction of actual cases, as under-reporting of sexual crimes remains a serious problem, especially for India's most vulnerable women. For example, extremely high rates of sexual assault and abuse of female sex workers in India have been recorded.[53] Also, the lack of sanitation facilities in India means that many women find themselves vulnerable to sexual assault.[54][55]
Sexual harassment, often called 'eve teasing', remains a significant problem in India. Following the high profile case of sexual harassment decided by the Supreme Court in 2009, India developed sexual harassment guidelines for workplaces, including the establishment of commissions that are at least 50% female to receive and act on complaints of sexual harassment.[56]The Sexual Harassment of Women at Workplace (Prevention, Prohibition and Redressal) Act was enforced on 9 December 2013.[57] In addition to civil penalties for sexual harassment in the workplace[58], the Criminal Law (Amendment) Act of 2013 created a new offense of sexual harassment inserted in the Penal Code,[59]and created voyeurism and stalking offenses.[60]
There is no evidence to suggest that female genital mutilation (FGM) is practised in India.
According to the US Department of State, so-called 'honour' killings remain a serious problem in India, particularly in the states of Punjab, Haryana, and Uttar Pradesh, with one report estimating that more than 1000 women and girls are killed this way each year.[61] Women and girls were killed for marrying or being in relationships without their families' or village elders' consent, or for marrying outside their caste.[62]
Abortion is legal in India in cases of foetal impairment, where the woman's mental or physical health is in danger, in cases of rape and incest, and for economic or social reasons.[63] Sex-selective abortions are a criminal offence.[64] However, significant obstacles to obtaining safe and legal abortion in India still exist, including: prohibitive costs; shortage of trained providers and adequate equipment; lack of confidentiality and informal demands for spousal consent; poor access to facilities; and lack of knowledge about the legal status among women, lawyers, and medical professionals concerning abortion and where to access safe services. [65] As a result, one study has found that of the 6.4 million abortions performed in India annually, 3.6 million, or 56%, were unsafe.[66]
Supreme Court rulings in cases concerning coercive and unsafe sterilization in India have mandated extensive guidelines for sterilizations, with an emphasis on counseling and informed consent.[67] Yet, implementation of these guidelines has been insufficient, and there have been several media reports of sterilization abuses, some resulting in death, throughout the country.[68]
Although India's Supreme Court has consistently held that maternal health is a constitutional and human right[69], in practicethese rights are often unfulfilled. Reproductive rights advocates have claimed that India continues to have the highest number of maternal deaths in the world due to poor quality of maternal health care, the prevalence of unsafe abortions, and barriers to accessing contraception.[70] The World Health Organization (WHO) estimates that 56,000 women and girls in India died from maternal causes in 2010.[71]
In some areas, access to reproductive health services is limited by lack of provision, although there are regional differences[72]Current use of contraception lags behind the level of knowledge, as roughly half of married women reported using a modern method in 2006.[73] Among women who did not intend to use contraception in the future, 42% did so because they had difficulty or an inability to become pregnant.[74] Overall, 12.8% of women had an unmet need for family planning, either because they desired to space the births of the children farther apart or because they wanted no more children.[75]
India is a source, destination, and transit country for women, and children subjected to forced labour and sex trafficking, with some NGOs observing that the level of human trafficking is increasing, especially from India's most disadvantaged social strata, including the lowest castes, which are most vulnerable.[76] According the US State Department, the Government of India does not fully comply with the minimum standards for the elimination of trafficking, and complicity of some government officials in human trafficking remained a serious and unaddressed problem that impeded efforts to adequately fight the crime.[77]In April 2013, the Government amended the Penal Code in a manner that broadened the types of crimes considered to be trafficking and establishing punishment ranging from 10 years to life.[78] The Ministry of Home Affairs continues to establish Anti-Human Trafficking Units, which are responsible for combining law enforcement and rehabilitation efforts. However, some of these local-level units were criticized as being ineffective or only established on paper.[79]The UJJAWALA programme offers support to victims of trafficking and commercial, sexual exploitation, with a focus on prevention, rescue and rehabilitation.[80]
[27] Arts. 18, 23, 24 [28] The Protection of Women From Domestic Violence Act 2005, Art. 5 [29] Amendment Act Art. 5, Penal Code Arts. 362A & 362B [30] US State Department (2012) [31] CEDAW (2005), p.7. [32] NCRB (2012) [33] IDHS (2006) [34] IIPS and Macro International (2007), Tables 14.15.1 and 14.15.2 [35] Ministry of Health and Family Welfare (2013). [36]Ministry of Women and Child Development (2013). [37] Lawyers Collective (2014). [38] http://nalsa.gov.in/ [39] http://www.itmd-india.org/index.php?option=com_content&view=article&id=42&Itemid=163 [40] Amendment Act Art. 9, Penal Code Art. 376[41] Amendment Act Art. 9, Penal Code Art. 376A [42] Amendment Act Art. 9, Penal Code Art. 376D [43] Amendment Act Art. 3; Penal Code Art. 166A [44] Amendment Act Art. 3, Penal Code Art.166B [45] Ministry of Law and Justice (2012), The Protection of Children from Sexual Offences Act, The Gazette of India, http://wcd.nic.in/childact /childprotection31072012.pdf [accessed 23/05/2014] [46] OHCHR (2013) [47] The Criminal Law (Amendment) Ordinance, art. 8, No. 13 of 2013, India Code (2013) [48] US State Department (2012) [49] Agnes, Flavia (2013). [50] NCRB (2012) [51] Sen, Samita (2013)[52] NCRB (2012) [53] Beattie (2010). p. 476 [54] DNA India (2014). [55] Akram, M. (2013). [56] UN Women (2011), p.17 [57] Lawyers Collective (2013). [58] The Sexual Harassment of Women at Workplace (Prevention, Prohibition and Redressal) Act, 2013 [59] Amendment Act Art. 7), Penal Code section 354A [60] Amendment Act Art. 7, Penal Code Arts. 354C & 354D [61] US State Department (2012) [62] US State Department (2012) [63] UN (2011) [64] The Pre-Conception and Pre-Natal Diagnostic Techniques (Prohibition of Sex Selection) Act, 1994 (PNDT), amended in 2003, cited in CEDAW (2005), p.17 [65] CRR (2012), p.3 [66] CRR (2012), p.3 [67] Ministry Of Health & Family Welfare, Manual For Family Planning Insurance Scheme 3-4 (2009), [68] CRR (2012), p.5 [69] See, e.g., Consolidated Decision, Laxmi Mandal v. Deen Dayal Harinagar Hospital & Others, W.P. (C) No. 8853/2008 & Jaitun v. Maternal Home MCD, Jangpura & Others, W.P. (C) Nos. 8853/2008 & 10700/2009 Delhi High Court (2010) (India) [70] WHO (2013) [71] WHO (2012) [72] CEDAW (2005), pp.76, 80; National Family Health Survey 3, p. 127 http://www.rchiips.org/nfhs/NFHS-3%20Data/VOL-1 /Chapter%2005%20-%20Family%20Planning%20(555K).pdf [73] UN (2012b); IIPS and Macro International (2007), Table 5.5 [74] IIPS and Macro International (2007), Table 5.23 [75] IIPS and Macro International (2007), Table 5.31 [76] US State Department (2013) [77] US State Department (2013) [78] Criminal Law (Amendment) Act 2013, Art. 370 [79] US State Department (2013) [80]Ministry of Women and Child Development (2007).
The male/female sex ratio for the working age population (15-64) in 2014 is 1.07 male(s)/female while the sex ratio at birth is 1.12.[81] There is strong evidence to suggest that India is a country of high concern in relation to missing women. UNDP reports that, as at 2008, India had a total of 42.6 million missing women, with numbers increasing in absolute terms.[82] 250,000 girls in India were thought to be missing at birth in 2008 alone.[83] The 2011 Census found a worrying trend in child sex ratios with only 914 females for 1,000 males, a drop from 927 in 2001. Using data from the 2011 Census in India, after adjusting for excess mortality rates in girls, the estimates of number of selective abortions of girls rose from 0 to 2.0 million in the 1980s, to 1.2 to 4.1 million in the 1990s, to 3.1 to 6 million in the 2000s. The study shows that the problem is in fact growing amongst the middle class which suggests that missing women cannot be attributed to poor socio-economic status.[84]
According to data from the 2006-2007 Demographic and Health Survey for India, 41.5% of girls and 45.3% of boys under the age of two had received all their vaccinations.[85] Under-five mortality rates were higher for girls than for boys (79.2 per 1000 live births for girls, 69.7% for boys), while malnutrition rates were equal or slightly higher for girls.[86] Given that in most contexts, rates of under-five mortality and malnutrition are higher for boys than for girls (due to physiological differences between male and female children), this would indicate bias towards sons in regard to early childhood care.
Primary and secondary school enrolment and attendance rates are lower for girls than for boys (according to UNICEF),[87] indicating some son preference in regard to access to education. The lack of provision for facilities for girls, such as toilets, reinforces this issue.[88] This disparity become more pronounced with increasing levels of poverty, with the World Bank finding that while boys and girls from the top income brackets participate in school at similar rates, there is a gender gap of almost five years in the bottom income quantile.[89]
[81] CIA (2014) [82] UNDP (2010), p.34 [83] World Bank (2011), p.15 [84] Jha et al (2011) [85] IIPS and Macro International (2007), Table 9.4 [86] IIPS and Macro International (2007), Table 7.3, 10.1 [87] UNICEF (2013) [88] (2008) Right to Education and Total Abolition of Child Labour. New Delhi: National Commission for the Protection of Child Rights. [89] World Bank, Figure 7
Women's equal access to land and other property are guaranteed by law and have been upheld in several court cases.[90] The Married Women's Property Act, 1974 (article 4) provides for married women's earnings to be their separate property. To encourage equal property registration, states have introduced policies that lower property tax rates for women.[91] The Hindu Succession Act of 2005 granted Hindu, Sikh, Buddhist and Jain women (who make up the vast majority of women in India) co-equal inheritance rights to ancestral and jointly owned property[92], and the same rights as sons, whether they are married or not, to reside in or seek partition of the family dwelling house.[93]The National Gender Resource Centre in Agriculture has been set up in the Department of Agriculture and Cooperation (DAC), Ministry of Agriculture to promote women's land access.
Nevertheless, NGOs report that weak implementation of the law has undermined the real impact of the reforms. District court rules have not been revised; officers are not held accountable for seeking female heirs; the law's implementation is not monitored.[94] In practice, a number of obstacles prevent women from accessing land and non-land assets, including discriminatory inheritance customary practices, limited legal literacy, discriminatory attitudes towards women's ownership and control of land in the family and community and discrimination in the allocation of land through state transfer programmes.[95] There is limited national data available on women's ownership of land and property, however one 2012 UN survey of women in two States found: only 12 percent of women respondents reported that they have or believe that they will inherit land; eight percent currently owned land; only four percent of formally titled plots in Muslim families named the woman; and women felt their plot access was most vulnerable if they were to divorce their husbands, had a falling out with their family, or their family incurred debt.[96] The National Mission for Empowerment of Women also promotes a model intervention project launched in many districts in India, to ensure the socio-economic development of women in rural areas: A Poorna Shakti Kendra (PSK) is established in villages and offers services to women at the grassroots level, including information on legal rights and entitlements, as well as training and capacity building on leadership.[97]
There are no legal restrictions regarding women's access to finance and credit, and women have equal rights to register a business.[98] In 2012, the Government of India reported that to improve women's access to the formal banking system, public sector banks were directed by the Reserve Bank to earmark 5% of their Net Bank Credit for lending to women. Credit to women from Net Bank Credit increased from 2.36% in 2001 to 6.29% in 2009.[99] However, in practice, access to finance in India is restricted by lack of access to bank accounts. For example according to the World Bank in 2011, only 26.5% of women held an account at a formal institution, compared with 43.7% of men; and only two percent of women used that account for business purposes, as compared with 6.1 percent of men. Unbanked women respondents cited a family member having an account as one of the reasons they did not possess an account.[100] In 2011 the World Bank also found that 6.7 percent of women obtained an institutional loan in 2011, contrasted with 8.6 percent of men. [101]Women's access to credit in India is also restricted in practice due to restricted control over collateral such as land.[102] The Government reported in 2012 that a number of micro-credit initiatives have been established to increase women's access to credit, several of which focus on minority women.[103] However, UNDP has reported that in parts of southern India, where competitive pressures have started pushing multiple micro-loans onto women regardless of the stability of their businesses. This has resulted in indebtedness and backlash.[104]
[90] CEDAW (2005), pp. 45, 82-83, 92, 94 [91] UNDP (2010), p.70 [92] Oxfam (2013), p1 [93] UNDP (2010) [94] Oxfam (2013), p.2 [95] Agarwal (2002) [96] UN Women (2012), p.5 [97] National Mission for Empowerment of Women (2014) [98] Constitution of India, art 19(1)(g) [99] CEDAW (2012), p.16 [100] World Bank (2011) [101] World Bank (2011) [102] Agarwal, B. (2002) [103] CEDAW (2012), p.8 [104] UNDP (2010), p.53
There are no legal restrictions on women's access to public space. Previous restrictions requiring unmarried women to provide their father's signature on passport application forms have been removed.[105]
Freedom of speech is generally respected in India, although there are some restrictions on freedom of assembly and association.[106]There is a very large and well-developed NGO sector in India, including a large number of vocal women's rights advocates and organisations. According to the 2005 CEDAW report, women's rights NGOs are active in empowering, mobilizing and organising women, challenging discriminatory and harmful practices such as sex-selective abortion and early marriage, raising awareness and advocating for policy change and for the repeal of laws that continue to discriminate against women, as well as in service delivery, particularly in regard to support to victims of gender-based violence, reproductive health services, and water and sanitation.[107] However, NGOs have reported that women's human rights defenders have been targeted for their work, particularly in relation to the rights of marginalized women.[108] Recently under the Nirbhaya Fund", transport specific programmes to ensure women's security and prevent violence against women are being promoted.[109]
Women and men in India have the same rights to vote and stand for election.[110] Until 2012, two of the top political positions, the Presidency and the Speaker of the House, were held by women. 62% of respondents to a 2007 poll believe that men and women are equally capable as politicians.[111] Concerning women's political voice, as of 2014, women held 11% of seats in the Lok Sabha (lower house) of Parliament, and 26% of seats in the Rajya Sabha (upper chamber).[112]
As per the 73rd and 74th Amendments to the Constitution, 33% of seats for panchayats in rural areas and urban local bodies are reserved for women.[113] In addition, in August 2009 Parliament approved an amendment to the national constitution to increase reservations in the panchayats (village councils) to 50%.[114] In March 2010, the upper house of Parliament passed another Constitutional amendment that would reserve one-third of the seats in the national and state legislatures for women, but the measure has drawn strong opposition in the Lower House.[115] However, UNDP has noted that women panchayat members in India have faced some huge obstacles to participation—some of which have resulted in torture and even death.[116]
India extends maternity leave benefits to all employed women for twelve weeks at 100% of their pay, paid for by their employer.[117] However the large number of women employed informally and in the agricultural sector (and not in the organized, public, or governmental sectors) means that many pregnant women are not covered by this benefit.[118] In addition, during the last two decades, the greatest increase in women's employment has been in those sectors where maternity leave provisions do not apply.[119]The 2011 Indira Gandhi MatrritvaSahyagYojna (IGMSY) is a Conditional Maternity Benefit Scheme for pregnant and lactating women offering conditional cash transfers for improved health and nutrition behaviour change. The scheme is being implemented in 52 selected districts on a pilot basis. Pregnant women who are 19 years old and above are entitled to a cash incentive of Rs. 4000 in three instalments to cover the first two live births.[120] Also, Janani Suraksha Yojana is a scheme aimed at reducing maternal and infant mortality rates and increasing institutional deliveries in below poverty line families. The scheme provides financial assistance of Rs. 500 per birth up to two live births to the pregnant women who are at least 19 years old and belong to the below poverty line households.[121]
A woman has equal rights to confer citizenship to her children in the same way as a man.[122]
[105] CEDAW (2005), p.92 [106] Freedom House (2013) [107] CEDAW (2005), pp.3, 4, 16, 78 [108] Asian Human Rights Commission (2013) [109] Ministry of Women and Child Development (2013), Nirbhaya Fund, http://pib.nic.in/newsite/erelease.aspx?relid=101674, [accessed 03/06/2014] [110] CEDAW (2005), p.53 [111] Pew Research Center (2007), Question Q.43 [112] IPU (2014) [113] UN (2012), p. 10 [114] US Department of State (2012) [115] Polgreen (2010), p. A4 [116] UNDP (2010), p.86 [117] ILO (n.d) [118] CEDAW (2005), pp. 7, 30 [119] Chandrasekhar, C.P. and Jayati Ghosh (2007) 'Women Workers in Urban India', www.macroscan.org [120] Ministry of Women and Child Development, (2011) [121]http://nrhm.gov.in/nrhm-components/rmnch-a/maternal-health/janani-suraksha-yojana/background.html [accessed 27/05/2014], [122] Citizenship Amendment Act, 2003, Arts. 3 and 4
Agarwal, B. (2002) 'Are We Not Peasants Too? Land Rights and Women's Claims in India', SEEDS, Population Council, http://www.popcouncil.org/pdfs/seeds/seeds21.pdf, (accessed 10 February 2014)
Agnes, Flavia (2013) 'No Short Cuts on Rape. Make the Legal System Work'. Economic and Political Weekly. Vol. XLVIII, Number 2.
Akram, M. (2013), Sanitation, Health and Development Deficit in India: A Sociological Perspective (), http://www.sociologyofsanitation.com/honble-guests/sessionspeakers/sanitation-health-and-development-deficit-in-india-a-sociological-perspective/ [accessed 10/06/2014]
Asian Human Rights Commission (2013), 'India: Punish the perpetrators of assault on women Human Rights Defenders in Lalitpur, Uttar Pradesh', 16 May 2013, http://www.humanrights.asia/news/urgent-appeals/AHRC-UAC-069-2013, (accessed 10 February 2014)
Basu, Srimati (2001) She Comes to Take Her Rights. Indian Women, Property, and Propriety. New Delhi: Kali for Women.
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subset: commoncrawl


text:
Sun, 19 Apr 2009 /
2 Attending
Crate diggers extraordinaire Dan Tyler and Conrad McDonnell sit down for a chat with RA's Dave Stenton.
RA's Peter Chambers catches up with Hotflush head Paul Rose, AKA Scuba, to chat about Iron Maiden, Guns N' Roses and even a little dubstep.
Burning Man sculptures, steam room breaks and a fake hotel: It's all part of what has made Wolf + Lamb one of the most exciting labels of 2009. RA's William Rauscher sweats it out to find out more.
RA picks the May festival highlights from across the globe.
RA catches up with the talented newcomer as he chases the dream of being able to make a living from the thing he loves the most: music.


text:
I had a mastectomy after being diagnosed with breast cancer 9 years ago at the age of 45.
In October 2011, after much consideration I decided to have a breast reconstruction using an abdominal flap. I preferred this procedure as it meant using my own body tissue instead of a foreign implant, looking and feeling more natural (I also liked the idea of a bonus tummy tuck). I found that the tummy area took a little longer than the breast to heal but I was pretty much back to normal after 6 weeks.
Since then I have had my other breast lifted and recently had a nipple reconstruction. After the surgery I feel normal again and a lot more confident. I can honestly say I am very happy with the results, I only wish I had it done sooner.


text:
"The conditions were very close to how it could be on the (Sept.) 3rd, it was very hot and humid at kickoff, but then it got cloudy and we got a bit of breeze, and you could tell a different energy level," said Richt.
The end of the scrimmage featured goal line situations which brings out the best of the players and coaches.
"It's high energy and comical at the end, everyone is having good, clean fun," said Richt. It's a mad celebration for one side depending on if the offense scores or the defense stops them. It's like a boxing match with everyone real close together watching to see which side wins."
The players and coaches were not the only ones watching closely Saturday as the afternoon scrimmage featured a crew of replay officials. The Southeastern Conference will implement the use of video replay this season, and they had their own scrimmage of sorts. Richt said he knew of a couple of plays that they reviewed, and the calls on the field were correct.


text:
Cameron Hayley, who has raced across the Pacific Northwest and Western Canada before joining NASCAR's K&N Pro Series full-time in 2012, won his biggest main event of his young career Tuesday night at Daytona International Speedway.
Courtesy: NASCAR Media Relations
DAYTONA BEACH, Fla. – Cameron Hayley came from behind during a green-white-checkered finish to win the inaugural UNOH Battle At The Beach NASCAR K&N Pro Series race Tuesday night at Daytona International Speedway.
It was only fitting that the third and final race of the two-day event concluded with cars trading paint, given that the first two races ended with last-lap contact and late lead changes.
Hayley took advantage when the race leaders tangled in Turn 2 to drive to the victory.
Fifteen-year-old Gray Gaulding finished second in the Krispy Kreme Toyota, while Puerto Rican Bryan Ortiz took the third spot in the Putinka Vodka Toyota.
"It was a tough race overall," said Hayley, who hails from Calgary, Alberta, Canada. "When I was sitting there on that last restart, I was thinking they were going to wreck in front of me. There's going to be something happening in front of me. My crew chief and my spotter coming over the radio telling me to be patient and when you see a hole - go."
Tuesday's nightcap, the feature of the UNOH Battle At The Beach, showcased 28 drivers from 11 different U.S. states, Canada, Mexico, Puerto Rico and Spain.
Other notables included Ben Kennedy, son of International Speedway Corporation CEO Lesa France Kennedy, who finished fifth. Kennedy is the first France family member to race at Daytona International Speedway in a stock car.
"It was a great experience to run at DIS, really cool what the track has put together for tonight," Kennedy said. "Unfortunately we got knocked around and messed up our rear end for the rest of the night. Really should've been in Victory Lane tonight but proud of the guys. We'll go on to Bristol and set our sights on the championship."


text:
Medical Definition of Dermis
Dermis: The lower or inner layer of the two main layers of cells that make up the skin.The dermis contains blood vessels, lymph vessels, hair follicles, and glands that produce sweat, which helps regulate body temperature, and sebum, an oily substance that helps keep the skin from drying out. Sweat and sebum reach the skin's surface through tiny openings in the skin that act as pores.
Last Editorial Review: 5/13/2016
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The American concussion expert who is conducting post-mortem analysis on the brain of an elite Australian rugby union player says a test to detect a head trauma-related disease on living athletes is ''very close''.
Chris Nowinski, the co-director of Boston University's Centre for the Study of Traumatic Encephalopathy, believes it would be ''naive'' to think that Australian footballers had not suffered from the same degenerative brain condition - chronic traumatic encephalopathy associated with memory loss, depression and early-onset dementia - that has been proved in US athletes and directly related to repeated head knocks.
America's National Football League agreed two weeks ago to pay $US765 million ($822 million) to settle a historic lawsuit over head injuries in the sport, and Nowinski has been a key figure in related research.
At present assessing the first brain of an elite Australian footballer at Boston University's ''brain bank'', Nowinski has told Fairfax Media of his plans to meet for a second time with Australian rules representatives on an imminent trip here. After a meeting with AFL Players Association general manager Ian Prendergast in the US last year, Nowinski described AFL as a sport with a ''very high risk for brain trauma''.
On the eve of the 2011 season the AFL introduced more conservative rules about diagnosing and managing concussion, rules that were bolstered this year with the introduction of a ''concussion substitute''.
The most alarming known concussion-related AFL cases have been outlined publicly by ex-Melbourne player Daniel Bell, who revealed in 2011 he'd been diagnosed with a brain injury after being concussed on as many as 25 occasions, and Brownlow medallist Greg Williams who this year detailed his concussion-related battles.
While there is no known proof of Australian footballers from any code suffering CTE, Boston University received the brain of ex-Manly player and coach Barry ''Tizza'' Taylor earlier this year. Nowinski is a member of the team assessing Taylor's brain for CTE and said the post-mortem analysis, which takes a minimum of two months, was almost complete.
Taylor's family would have to approve the release of the findings.
While tests for CTE can only be conducted post-mortem, Nowinski has forecast the emergence of a test for living humans ''in the next year or two''.
''There's been more funding allocated to CTE research in the last year than in the history of the world, I think. It's exciting to see that more researchers are working on this because it's a big problem,'' he said.
''I think we're very close to testing for it [CTE] while someone's alive.
''We're collaborating with a number of people on ways to diagnose it in living people, so it would not surprise me if in the next year or two athletes can know if they have CTE while they're alive. It will be a huge breakthrough.''
There are more than 50 brains of former NFL players in the Boston brain bank and about 172 brains of former athletes - including Taylor's - in total.
''There's no reason to believe that we wouldn't find CTE in former Australian football players if we looked, because we're adding sport after sport where we're finding the disease,'' Nowinski said.
AFL Medical Officers Association boss Dr Hugh Seward said on Thursday there would be a review of the league's concussion guidelines at a conference in November.
Seward said he would be interested in meeting Nowinski again - the pair spoke at an international concussion summit in Zurich last year - during his trip to Australia.


text:
What the Nobody looks like in the lower realm
|Anonymous Coward
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Quoting: Anonymous Coward 17544037
Quoting: Anonymous Coward 26175223
sodom and gomorrah, it's coming, yesQuoting: Anonymous Coward 26096364
this OP is giving you a preview of that which will follow as the whole world falls into abomination with the entrance of the deceiver known as antichrist, who will be arriving at the sixth trumpet in "ships" from the skies, hence termed "aliens" by those who are unaware of the prophecies in this regard. it will be satan and the fallen angels, disguised as emissaries of "the light" who will be cast out and down onto the earth to tempt and deceive you for a five month period known as the "tribulation of the saints" and/or "time of the locust"
all those who wish to "overcome" and put on immortality at the actual second advent, which occurs FIVE months after their arrival, exactly five months, yes...
must refuse to worship them, must stand against them accordingly
they will offer you bribes, new technologies, prosperity, you name it. they will tell you that you ARE all GODS and that there is no such thing as "sin" and you will be sorely tempted accordingly, but it's all lies, and once they arrive the clocks ticking, only five months left before the end and all are changed back into spiritual bodies as the flesh falls away. (think galactic federation channeled entities, if this isn't these they will be identical in form and format)
either way it won't be an easy row to hoe. be in tribulation with the supernatural antichrist aka satan and the fallen angels...
or be in tribulation from the Father who created all the aforementioned entities, and who owns all of Creation, including your mortal souls.
five months with antichrist - 1000 years in the Millennium, cooling your heels on the sad and sorry side of the gulf that will separate you from overcoming, from the fully resurrected souls, from Christ, from the Temple, and from the Father...
choice is your own. nobody can rue your choices for you but yourselves, and/or take pride in them accordingly, after the fact.
by the way, at the actual second advent the fallen angels will be destroyed, permanently, poof
at that time satan will be tossed back into the pit and you will be able to see him there during the Millennium Age. gaze in upon him and wonder how in hell that which lies before you could have tempted you in any way, shape, or form.
he won't look so high and mighty from that angle, trust me
tho neither will you from the wrong side of the gulf.
god is so strange. Why create a being who you know will deceive other beings you created only to throw them in a hell you created as well? Why bother creating them in the first place?
Why bake a cake? It's going to make a mess and you're going to have to wash dishes and take out the trash afterwards...Why even do it at all?
So god is not perfect? Can he not create a cake without making a mess?
|Anonymous Coward
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The nobody, being a shapeshifter, can assume one of any of the characters/actors from Diff'rent Strokes, the television show of the 1980s era.Quoting: facial 25676746
When he wants to symbolize the plight of the second son, or the inner city feelings, he will look like Willis Drummond.
When he wants to merge with the elites and visit penn state and other places where you have to look a certain way, he appears as the Dad, Mr. Drummond.
When he wants to invoke Goddess power, he appears as Kimberly Drummond.
And finally, when he wants to be himself and have a form that is magically acceptable to all advanced civilizations, he assumes the form of Arnold, the loveable pint size wise cracker.
There is some other TV show he can assume forms from, it could be Battlestar Galactica but I'm not sure. Rumor has it he once formed into a 10 foot tall Kermit the Frog, but since none of those present had birth certificates or drivers licenses, the authorities said the was all drunk on moonshine.
The 'Nobody' may be a shapeshifter at the level of his consciousness; but that's about it while physiologically embodied.
|ChemicalEngineer (OP)
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|Anonymous Coward
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|What is the lower astral realm?
|CONFIRMED: The Kingdom of Heaven is a REALM that works like a COMPUTER!
|An interesting spiritual trip i had. Reincarnation Universal Timeline Lower Realm
|Has anyone ever seen a mind? What does a mind look like? Can you see the immaterial realm where the soul resides?
|Putin in turkey to stabilize the caspian energy realm...imin zawahiri does not like this


text: 9/05/08 3/07/08 POPSUGAR Fashion POPSUGAR Celebrity Lorick Fall 2008 Look Book March 7, 2008 by coutorture 0 Shares Chat with us on Facebook Messenger. Learn what's trending across POPSUGAR. View on One Page Photo 0 of 33 Start Slideshow ADVERTISEMENT () Share This Link Copy What's Your Reaction? Thanks for your reaction Don't forget to share this with your friends! ← Use Arrows Keys → 0 Lorick Fall 2008 Look Book Archetype Showroom Previous Next


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ISRN Biochemistry
Volume 2013 (2013), Article ID 910308, 10 pages
Evaluation of the In Vitro Efficacy of Artemisia annua, Rumex abyssinicus, and Catha edulis Forsk Extracts in Cancer and Trypanosoma brucei Cells
1Institute of Biochemistry, Faculty of Medicine, University of Leipzig, Johannisallee 30, 04103 Leipzig, Germany
2Institute of Public Health, College of Medicine and Health Sciences, University of Gondar, P.O. Box 198, Gondar, Ethiopia
3Physiology Department, Medical Faculty, Jimma University, P.O. Box 378, Jimma, Ethiopia
4Department of Tropical Medicine, Medical Mission Institute, Salvatorstrasse 7, 97067 Würzburg, Germany
5Department of Veterinary Parasitology, Faculty of Veterinary Medicine, University of Leipzig, An den Tierkliniken 35, 04103 Leipzig, Germany
6Department of Zoology, Faculty of Science, University of Alexandria, Maharram Bey, Alexandria 21511, Egypt
Received 29 May 2013; Accepted 13 August 2013
Academic Editors: S.-P. Scott and N. Sergeant
Copyright © 2013 Netsanet Worku et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
The current drugs against sleeping sickness are derived from cancer chemotherapeutic approaches. Herein, we aimed at evaluating the in vitro effect of alcoholic extracts of Artemisia annua (AMR), Rumex abyssinicus (RMA), and Catha edulis Forsk (CEF) on proliferation/viability of 1321N1 astrocytoma, MCF-7 breast cancer, THP-1 leukemia, and LNCaP, Du-145, and PC-3 prostate cancer cells and on Trypanosoma brucei cells. Proliferation of tumor cells was evaluated by WST-1 assay and viability/behaviour of T. brucei by cell counting and light microscopy. CEF was the most efficient growth inhibitor in comparison to AMR and RMA. Nevertheless, in LNCaP and THP-1 cells, all extracts significantly inhibited tumor growth at 3 μg/mL. All extracts inhibited proliferation of T. brucei cells in a concentration-dependent manner. Microscopic analysis revealed that 95% of the T. brucei cells died when exposed to 33 μg/mL CEF for 3 hrs. Similar results were obtained using 33 μg/mL AMR for 6 hrs. In case of RMA, however, higher concentrations were necessary to obtain similar effects on T. brucei. This demonstrates the antitumor efficacy of these extracts as well as their ability to dampen viability and proliferation of T. brucei, suggesting a common mechanism of action on highly proliferative cells, most probably by targeting cell metabolism.
1. Introduction
Many anticancer reagents, including nucleotide analogues and other DNA synthesis inhibitors (e.g., methotrexate), aim at exerting a specific activity against rapidly proliferating cell types. Because of the capacity of trypanosomes for rapid growth within mammals, they have been linked to some types of cancer cells. D,L-α-Difluoromethylornithine (DFMO), a polyamine synthesis inhibitor, was originally developed as a potential anticancer reagent before it has been proved useful in trypanosomiasis therapy. Furthermore, suramin, melarsoprol, and pentamidine which are drugs licensed for the treatment of human African trypanosomiasis (HAT) are known to arise from cancer research studies [1–3]. If left untreated, sleeping sickness patients die within months when infected with T. brucei rhodesiense (acute form of the disease in East and Southern Africa) or within years when infected with T. brucei gambiense (chronic form of the disease in West and Central Africa) [4]. It is estimated that at least people are presently infected [5]. Above this burden of the disease, all the available drugs have a number of shortcomings during treatment because of resistance development against them, allergic reactions, undesirable effects in the urinary tract, and reactive encephalopathy (with 3–10% fatality), and some regimens are strict and difficult to apply [6]. This raises an urgent need for novel, safe, rapidly-acting, and inexpensive agents for the treatment of HAT [7].
Our project selects three plants called Catha edulis Forsk (CEF), Artemisia annua (AMR), and Rumex abyssinicus (RMA), which are locally endemic to Ethiopia and widely used in the community for various purposes. The pharmacological effect of the selected plants on different cell lines and the known similarity of chemotherapeutics against trypanosomes and cancer initiated us to continue investigation of drugs to be commonly effective against these two important diseases.
CEF leaves were commonly used as a stimulant; that was formerly believed to be due to its norpseudoephedrine content. However, it might be cathinone, a newly discovered potent alkaloid with a pharmacological profile closely resembling that of amphetamine, which is responsible to this effect [8]. It can be said that CEF is an amphetamine-like plant material, the effects of which can be accounted to the "khatamines" including phenethylamines cathinone and, to a lesser extent, norpseudoephedrine. In addition to these two "khatamines," the leaves also contain the cathedulins [8],an interesting group of substances awaiting pharmacological elucidation.
AMR contains artemisinin, an endoperoxide consisting of sesquiterpene lactone [9]. Unlike other peroxide compounds, artemisinin can be stored at room temperature for many years without noticeable decomposition. Semisynthetic derivatives include artemether, arteether, and artesunate [10]. Interestingly, artemisinin is essentially nontoxic to normal cells [11]. Zheng [12] reviewed the cytotoxic activity of artemisinin, and recently it has been shown to be effective in killing cancer cells by inducing apoptosis and inhibition of nuclear factor κ-B, an important activator in cancer development and progression [13, 14]. With regard to its antiprotozoal effect, artemisinin has shown a promising antileishmanial activity [15], and currently an artemisinin-based combination therapy (ACT) is recommended for the treatment of Plasmodium falciparum malaria [16, 17].
RMA is widely used in folklore medicine for treatment of headache, hemorrhoid, ascaris, scabies, leprosy, fungal skin infection, wounds, eczema, and sore-throat and to control mild forms of diabetes. The decoction of the root or leaf powder of the plant is used as vermifuge [18, 19]. Studies have also shown its effect against leukaemia cells by initiating cell death through apoptosis [20]. Unlike the other species of this plant, studies done on the effect of RMA against cancer cells, however, are preliminary.
The objective of this study was, therefore, to evaluate the in vitro effect of the three aforementioned plant extracts against cancer cells and Trypanosoma brucei cells and to determine the prospect of their common application against these two respective diseases.
2. Materials and Methods
2.1. Mammalian Cell Culture
Six different tumor cell lines from prostate cancer (PC-3 (CRL-1435, ATCC), Du-145 (ACC 261, DSMZ), and LNCaP (ACC 256, DSMZ)), brain astrocytoma 1321N1 cells (86030102, ECACC); breast cancer MCF-7 cells (ACC 150, DSMZ), and leukemia THP-1 cells (LH-1, ATCC) were cultured in RPMI medium in the absence of fetal calf serum (RPMI-SF) or the presence of 10% fetal calf serum (RPMI-FCS). PC-3 cells, however, were cultured in Dulbecco's modified Eagle's medium in the presence of 10% fetal calf serum (DMEM-FCS). All growing media contain penicillin/streptomycin (100 U penicillin/mL; 100 μg streptomycin/mL), and cells were incubated at 37°C/5% CO2. The overall viability of cells used was in the range of 90% to 95%.
2.2. Trypanosoma Cell Culture
Blood stream forms (BSF) of T. brucei brucei laboratory adapted strain (TC-221) cells were cultured in complete Baltz medium, containing 82% Baltz medium basic solution, 0.8% β-mercaptoethanol stock solution (20 mM), 0.8% penicillin/streptomycin (10,000 U/mL), and 16.4% heat inactivated FCS all from Biochrom, Berlin, Germany. The 500 mL Baltz medium basic solution contained 3 gm HEPES, 500 mg glucose-monohydrate, 110 mg sodium pyruvate, 7 mg hypoxanthine, 2 mg thymidin, 10.7 mg adenosine, 14.1 mg bathocuproine sulfonate, and 146 mg of L-glutamine, all from Roth (Karlsruhe, Germany) except bathocuproine sulfonate that was purchased from Sigma Aldrich (Seelze, Germany). These compounds were dissolved in 500 mL modified Eagle's medium to which was added 5 mL of nonessential amino acids (Biochrom, Berlin, Germany).
2.3. Plant Extracts
Three different medicinal plants (Catha edulis Forsk, Artemisia annua, and Rumex abyssinicus) were deposited in and identified by the National Herbarium, Faculty of Science, Addis Ababa University in Ethiopia. The plants were selected mainly on the basis of their extensive usage in the community and/or on their local medicinal knowledge. The air-dried and powdered plant materials were extracted with 95% methanol at 60°C for 8 hrs using Soxhlet apparatus. The obtained methanol extract was vacuum-filtered and evaporated by using a rotary evaporator and freeze-dried (D. Piatkowski, Munich, Germany). The dried extracts were stored at −20°C until used. Stock solutions were prepared in DMSO just prior to the start of experiments so that its final concentrations did not exceed 1% during the experiment.
2.4. Tumor Cells Vitality Test
Proliferation/vitality of tumor cells was assessed using the WST-1 assay according to manufacturer's instructions. This colorimetric assay was used to determine cell proliferation/cytotoxicity in 96 well plates (5000 cells/well). Increasing concentrations of plant extracts (3 to 333 μg/mL) were added after preincubation of cells for 24 hrs (start value). A volume of 12 μL stock solution of WST-1 was added to each well plate, and was incubated for 4 hrs at 37°C. As a control, DMSO was added instead of the plant extracts. The absorbance was directly recorded on an ELISA reader/96-well multiscanner at a wavelength of 450 nm.
2.5. Impact of Exposure of Plant Extracts on Survival of Trypanosoma brucei Cells
Time dependency test was done in 24 well plates where 1 mL of cell suspension ( cells) plus 100 μL of plant extracts giving a total volume of 1.1 mL/well were incubated for 24 hrs at 37°C in humidified environment containing 5% CO2. To count the cells first the plate was slowly shaken in circular motion and well mixed using a 100 μL pipette to help the cells float well in the media just before pipetting for counting on a Neubauer hemacytometer. Ten microliters of the cell suspension has been taken, and the number of moving cells was then recorded after 1 hr, 3 hrs, 6 hrs, and 24 hrs of incubation. Tests have been performed in triplicates, and the average values were obtained for two repeated experiments (). The controls were seeded in a 1.1 mL of fresh medium containing 1% DMSO and handled in exactly the same way as the exposed cells.
2.6. Microscopic Observation of Trypanosoma brucei Cells' Viability
This test was done using 24 well plates containing 1 mL of cell suspension ( cells) plus 100 μL of plant extract. Cells were exposed to three different concentrations of the respective plant extracts, and the activity of cells was evaluated by monitoring (i) movement of live cells, (ii) morphology of the moving and nonmoving cells, and (iii) the number of live moving cells recorded at the 2nd, 3rd, 6th, and 24th hr of incubation at 37°C in humidified environment containing 5% CO2 and compared with the controls.
2.7. Statistical Analysis
Count and percentage of dead/viable cells has been estimated to compare effect of plant extracts on exposed cells and the corresponding controls. The optical density reading was used to determine the relative rate of cell proliferation to determine the IC50 of plant extracts using a linear interpolation. The statistical significance of difference of two means in cancer cells experiment was calculated using unpaired -test, and values of were considered to be statistically significant. The mean and standard deviation values are also employed for comparison. The data analysis, all figures, and tables presented were done using GraphPad Prism (version 4) statistical software.
3. Results
3.1. In Vitro Evaluation of Anticancer Effect
CEF showed concentration-dependent inhibitory effect in all cancer cells tested, and a statistically significant inhibition of proliferation () was observed at 3 μg/mL concentration in LNCaP, astrocytoma 1321N1, MCF-7, and THP-1 cells. When compared with the other cancer cell types tested, the lowest recorded IC50 was found in case of LNCaP prostate cancer cells (2.4 μg/mL). In addition to LNCaP prostate cancer cells, CEF best inhibits THP-1 leukemia and MCF-7 cells when applied at concentrations ≥33 μg/mL. In all cases, the maximum concentration applied resulted in nearly 100% inhibition. In PC-3 cells, however, maximum constant inhibition has been reached at 66 μg/mL (Figure 1).
AMR exerts a concentration-dependent effect on all cancer cells undertested except in PC-3 cells in which case it looks to have a provital effect at lower concentrations ≤33 μg/mL (Figure 2). In LNCaP, 1321N1, and THP-1 cells it revealed a statistically significant inhibition () at a concentration starting from 3 μg/mL. In the remaining three cancer cell types, however, a significant inhibition has reached concentrations greater than or equal to 100 μg/mL. The lowest IC50 (3 μg/mL) has been recorded in LNCaP cells when compared with the remaining cancer cells employed in our experimental study (Figure 2).
In case of RMA, the concentration-dependent inhibitory effect has also been well documented against growth of all cancer cell lines. A statistically significant growth inhibition () was observed at 3 μg/mL in LNCaP, 1321N1, and THP-1 cells. Unlike the other two plant extracts, however, the degree of growth inhibition by Rumex abyssinicus extract was low as it has not resulted in a complete inhibition of multiplication even at the maximum concentration (333 μg/mL) in all cancer cells tested. The lowest IC50 (29 μg/mL) was recorded in THP-1 cells. In these cells the maximum dilution had resulted in only 70% inhibition (Figure 3).
3.2. In Vitro Evaluation of Antitrypanosomal Effect
The proliferation of T. brucei cells has been assayed using Alamar blue, an indicator for metabolic cell function, evaluated as a colorimetric dye. However, when the Alamar blue was mixed with the growth media, the deep red or yellow coloration of the extracts at higher concentrations interferes with the absorbance measurement. For this reason, we have employed the time dependency test by counting the number of actively moving cells after exposing the cells to the plant extracts for 24 hrs at 37°C in 24 well plates. Cell counting has been done at the start of seeding, and at 1 hr, 3 hrs, 6 hrs and 24 hrs incubation periods. Microscopic observation on the relative speed of cellular motility and their morphological changes has also been recorded.
Results demonstrated that CEF inhibited proliferation of T. brucei cells at the last two higher dilutions (166 and 333 μg/mL), and its initial effect was observed within the first hour of incubation, and its maximum effect was seen at the 6th hr (Figure 4(a) and Table 1). At concentrations of 3 and 8 μg/mL, no inhibitory effect was observed. At 33 μg/mL, however, cytostatic effect was observed where the number of cells remains almost constant within the following 24 hrs of incubation (Figure 4(a)).
AMR similarly inhibits growth of cells at concentrations of 33 μg/mL, 166 μg/mL, and 333 μg/mL. The inhibitory effect was detected following the first hour of incubation and continues until it kills all the cells within the next 24 hrs of incubation (Figure 4(b) and Table 1). At 3 μg/mL and 8 μg/mL concentrations, however, no inhibition had been observed whereby it resulted in a cytostatic effect at 8 μg/mL, as the number of cells kept constant over a time elapse of 24 hrs (Figure 4(b)).
RMA also results in 100% inhibition at ≥166 μg/mL after 24 hrs of incubation. At 33 μg/mL, the cell number had decreased to a value of about 30%, but no inhibitory effect at 3 or 8 μg/mL was observed (Figure 4(c)). Table 1 lists the phenotypic changes recorded upon monitoring cells exposed to the various concentrations of CEF, AMR, and RMA extracts.
4. Discussion
Although cancer is not a single disease, and Trypanosoma are eukaryotic parasites presenting a high level of genotypic variability, the growth of the parasites in their host and that of cancer cells share at least one common feature, that is, their mutual capacity for rapid cell division [21]. Such cells require high polyamine concentration, and the inhibition of polyamine metabolism highly affects their survival. For example, the drug eflornithine (DFMO) is known to affect the polyamine synthesis in both Trypanosoma and cancer cells.
Another area of interest linking these two cell types is the glycolytic pathway. Tumor cells gain their energy mainly from aerobic glycolysis, and trypanosomes solely depend on glycolysis which is critical to their survival. For example, silencing the hexokinase 1 (TbHK1) gene in T. brucei induced parasite death, and a similar effect has been observed using the hexokinase inhibitor lonidamine, both in cancer and trypanosome cells [22].
For the treatment of sleeping sickness only four drugs, of which three were developed 50 years earlier, are available. Eflornithine (DFMO), a selective inhibitor of ornithine DL-decarboxylase, is the only new drug for chemotherapy of sleeping sickness. It was first used in 1990 and is only effective against T. b. gambiense [23]. With the exception of the prodrug pafuramidine maleate (DB289), an orally bioavailable diamidine about to enter Phase II clinical trials [24], no other compounds are in clinical development, indicating that no new antisleeping sickness drug will become available in the next few years [25]. In the light of declining investment in drug development for parasitic diseases, new strategies for the chemotherapy of HAT are urgently required [26]. Among these strategies it is promising to employ drugs developed as potential antitumour agents in the fight against HAT. The current antitrypanosoma drugs including suramin, pentamidine, melarsoprol, and eflornithine are good examples supporting this strategy [23, 25–27]. Similarly, (−)-epigallocatechin-3-gallate, a constituent of green tea, has antioxidant, anti-inflammatory, antimicrobial, anticancer, and antitrypanocidal activities [28]. Investigation of other plants with antitrypanosoma and anticancer activities is the easiest way to evade the long and costly process of development of new antisleeping sickness drugs. Here we, therefore, present the pharmacological activity of plants Artemisia annua, Rumex abyssinicus, and Catha edulis Forsk against T. brucei and different cancer cells.
By comparing the efficacy of these three plant extracts in inhibition of proliferation of cancer cell lines, we have observed that CEF was more effective than AMR and RMA, showing a statistically significant inhibition in four of the cancer cell types at the lowest concentration used (3 μg/mL). On the other hand, when the complete inhibitory effect of the maximum dilution was considered, RMA was not as effective as AMR and CEF. Among all cell lines tested, LNCaP prostate cancer and THP-1 leukemia cells were found to be most susceptible to the inhibitory effects of all the three plant extracts, where their IC50s lay below 33 μg/mL. In addition, it was observed that extracts of CEF, AMR, and RMA displayed antiproliferative effects on T. brucei cells. Best inhibitory effects were observed at concentrations ranging between 33 μg/mL and 333 μg/mL. Similarly, T. brucei cells proved to be most susceptible to CEF and AMR (Figure 4 and Table 1). As it was observed in a microscope, the effect of the latter two extracts was detected after 2 hours of incubation at concentrations ≥33 μg/mL (Table 1). Overall, all the three plant extracts exerted their inhibitory effect, even though with variable efficacies, both in cancer and trypanosome cells. This result is in accordance with previous findings using various cancer cell lines and trypanosome species [8, 29–31].
Our study on T. brucei cells has demonstrated that 98% of cells died after 6 hrs at 33 μg/mL of the CEF extract (Table 1). A study done by Dimba et al. [32] described a novel mechanism of CEF-induced cell death by apoptosis involving induction of caspases in HL-60 leukemia cell lines. They had also shown that its effect is rapid resulting in cell death within 2 hrs. A similar study done by Rosenkranz and Wink [33] has also shown that alkaloids induce apoptosis in blood stream form T. brucei. Alkaloids are known to affect protein biosynthesis, interact with DNA, and disturb membrane fluidity and/or microtubule formation. This might be the reason for the rapid cell death observed in T. brucei cells in our study. However, the type of cell death process in our experiments must be clearly demonstrated in future studies.
The current experimental study similarly had shown a good inhibitory effect of CEF on THP-1 leukemia cells, starting from 33 μg/mL, which is in agreement with the study done by Dimba et al. [32]. In the other cancer cell lines used in the current study, CEF extract efficiently inhibited cell proliferation even at the lower concentration, supporting its activity as an anticancer and antitrypanosomal drug. This result, therefore, is in agreement with in vitro antimicrobial property studies against periodontal pathogens done by Al-Hebshi et al. [34].
As reviewed by Sinclair et al. [35] on the pharmacological effect of artemisinin on cancer cells, it is indicated that its mechanism of action is done by affecting the transferrin receptor in iron metabolism that is pertinent for survival in almost all cancer cells. Transferrin receptors are highly expressed in cell surfaces of breast cancer, brain tumour glioblastoma and meningo-glioma as well as in myelogenous leukaemia (CML) cells compared to their corresponding normal cells. Leukaemia cells were found to be most sensitive to artesunate. Accordingly, our result shown in Figure 2 was partly in support of these findings whereby a significant cell growth inhibition was observed both in 1321N1 and THP-1 cells at the lowest applied concentration. The MCF-7 cells were also affected significantly, but at a higher concentration than in case of the previous two cell lines. This warrants further investigation to identify the reason for their lower susceptibility albeit the high expression of transferrin receptors on their surfaces.
Artemisinin contains an endoperoxide bridge (R–O–O–R′) that interacts with Fe (II) to form toxic free radicals and is required for its antimalarial as well as anticancer activity. It is believed to exert its cytotoxic effect via Fe (II)-mediated cleavage of the endoperoxide bridge [11, 36]. An intact endoperoxide is crucial since artemisinin derivatives lacking an endoperoxide bridge are devoid of antimalarial activity [17]. In the case of bloodstream forms of T. brucei, iron is delivered by host transferrin to a heterodimeric, glycosylphosphatidylinositol-anchored transferrin receptor [37]. The trypanosomal transferrin receptor is homologous to the N-terminal domain of the variant surface glycoprotein (VSG) and bears no structural similarity with the human transferrin receptor [37]. This structural difference to the human transferrin receptor makes this pathway a good target to develop chemotherapy against human trypanosomiasis. A study done by Mishina et al. [38] also confirmed the inhibitory effect of artemisinin in the in vitro growth of T. cruzi and T. b. rhodesiense in low micromolar range. It targets the calcium dependent-ATPase activity in T. cruzi cell membrane. With respect to this fact, the strong efficacy of AMR to inhibit T. brucei cells' proliferation revealed during the current study is quite appreciated, where nearly 50% of cells died within 3 hrs of incubation (Table 1) and 100% after 24 hrs of incubation at 33 μg/mL (Figure 4(b)).
An in vitro experiment done by Maksimović et al. [39] on the fruit of the yellow dock Rumex crispus indicated that it has a specific antioxidant effect. It has also been identified that ethyl acetate extracts from the roots of Rumex dentatus contained four spectroscopically separable compounds and two of which were novel. From these four identified compounds chrysophanol exerted a better inhibition of growth in MCF-7 cell lines [40]. In the current study, Rumex abyssinicus selectively showed better inhibition in THP-1 and LNCaP cells that corresponds with the study done by Wagiera et al. [20] using Rumex dentatus. However, our findings did not agree with regard to the effect of this plant extract on MCF-7 cells when compared with the study done by Zhang et al. [40]. Otherwise, our study revealed that methanol extract of Rumex abyssinicus was not a good candidate of therapy mainly against MCF-7 and PC-3 cells, where by a statistically significant inhibition () has been seen only at the maximum concentration applied (333 μg/mL).
5. Conclusions
Our present study demonstrates the efficacy of CEF, AMR, and RMA in inhibiting growth of different cancer cell lines as well as T. brucei cells. The CEF extract was relatively the best effective extract in comparison to AMR and RMA against proliferation of all cell lines. In all cases, LNCaP and THP-1 cells were found to be the most susceptible cell lines among others. It has been recalled that almost all of the drugs against human African trypanosomiasis are derived from drugs for cancer. Our current results will also strengthen this consent and can be applied towards developing drugs to be commonly used in sleeping sickness and cancer. This area of integrated "cancer and neglected tropical diseases (NTDs)" research should further be promoted so that NTDs management suffering from various problems like drug resistance, insecticide resistance of transmitting vectors, and financial shortage of production of new drugs can probably be solved.
The authors greatly acknowledge the technical support of Ms Angelika Schäfer from the Institute of Biochemistry, Faculty of Medicine, University of Leipzig, and Ms Antje Fuss from the Department of Tropical Medicine at Medical Mission Institute in Würzburg, Germany. Netsanet Worku and Andualem Mossie highly acknowledge the scholarship supported by the German Academic Exchange Service (DAAD).
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text:
Blues coach Pat Lam will join up with his players a day after they arrive in Canberra for Saturday's Super Rugby match against the Brumbies, so he can attend an interview to remain in the job.
Lam is looking to extend his four-year tenure, despite a tough season in which the Blues are languishing near the foot of the table with one round to go.
He says the timetable will not be a distraction from preparations for a match the Blues need to win to ensure they avoid finishing with the wooden spoon.
He says most of the work has been done, with the final training run yesterday and the team leaving today. Lam will fly over after his interview.
Blues chairman Gary Whetton declined to say how many candidates had been shortlisted, saying that was confidential.
The New Zealand Herald reported that it understood there were four names on the list - Lam, former All Blacks John Kirwan and Kieran Crowley, who have both coached national teams overseas, and Sharks coach John Plumtree.
Meanwhile, the Blues have made three changes to their starting line-up from their 32-9 win over the Western Force last weekend. Tighthead prop Charlie Faumuina is back from two months out after tearing a calf muscle just days before being named in the All Blacks wider training group.
Chris Lowrey comes in as blindside flanker, with Steven Luatua moving to lock, while yet another injury means Hadleigh Parkes fills in at left wing.
Parkes replaces Rene Ranger, who is out with a groin strain, making him the 16th player from the Blues' original squad to be unavailable.
Side to take on ACT Brumbies on Saturday: Charles Piutau, Lachie Munro, Francis Saili, Ma'a Nonu, Hadleigh Parkes, Michael Hobbs, Piri Weepu, Luke Braid, Daniel Braid, Chris Lowrey, Steven Luatua, Liaki Moli, Charlie Faumuina, Keven Mealamu, Tony Woodcock. Res: Tom McCartney, Tevita Mailua, Andrew Van der Heijden, Peter Saili, Alby Mathewson, Orene Ai'i, Sherwin Stowers.


text:
AMHERST, Massachusetts -- Todd Nichols (Skaneateles, NY) was named the Defensive Player of the Week by both the New England Small College Athletic Conference (NESCAC) and the Eastern College Athletic Conference (ECAC) for his performance in a 22-17 victory over Middlebury yesterday. The Lord Jeffs are currently 3-0 for the 1997 season.
Nichols posted his second three-intereception game against the Panthers, picking passes deep in Middlebury territory. The free safety registered a five-yard return on his first interception to the Middlebury 13, from which the Jeffs scored for a 15-0 lead late in the second quarter. Shortly after halftime, Nichols downed his second pick in the endzone to shut out the Panthers in the third quarter. Two possessions later, the Political Science major spoiled another Brian Coates (Brookfield, CT) attempt at the Panther 45.
The junior now has eight career interceptions as a three-year starter. The last time Nichols posted a three-interception game was in a 24-7 victory at Trinity College in Hartford, Connecticut last November. His efforts had helped Amherst end the seven-year reign by the Bantams over Lord Jeff football.
In addition to disrupting the passing game, Nichols recovered a fumble with a 15-yard recovery early just 10:12 after the opening whistle. He also posted five tackes (three solo) and broke up a Panther pass. As a unit, the defense allowed Middlebury only 11-yards rushing as Amherst leads the NESCAC in both yards allowed (481) and point allowed (17).
Teammate Darin Byrne (Winchester, MA) also received ECAC recognition on the New England Division III Honor Roll. The running back catalyzed the Amherst offense with 29 carries for 129 yards and a pair of catches for 48 yards. The junior scored his first two touchdowns of the season with a 32-yard reception and a 1-yard run in the second quarter. Byrne is currently Amherst's leading rusher, averaging 74-yards per game and 4-yards per carry. The Jeffs compiled 152 yards rushing and 372 total against the Panthers, upping Amherst's three-game total to 1,157.
The Jeffs travel to Waterville, Maine on Saturday, October 11 for a meeting with Colby College. Last year Amherst blanked the White Mules 34-0 in Massachusetts. Amherst leads the series 15-4-1.
# # # 10/5/97


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