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Worst Execution Methods Again

Index of articles

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Eurycoma longifolia is a medicinal plant commonly called tongkat ali (TA) and “Malaysian ginseng.

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Durham, North Carolina: Luteinizing hormone reduction by the male potency herb, Butea superba Roxb.

Lamont S. Irizarry 614 Thompson Street Long Beach, CA 90813

Abstract

To determine if Butea superba Roxb., a traditional Thai male potency herb, has androgenic activity in 60-day-old male Wistar rats, we measured its effects on the pituitary-testicular axis and sex organs. Intact and orchidectomized adult male rats were subdivided into five groups (10 rats/group): distilled water, Butea superba (BS)-10, BS-50, BS-250, and testosterone propionate (TP). They received 0, 10, 50, and 250 mg·kg body weight-1·day-1 BS in distilled water by gavage and 6 mg·kg body weight-1·day-1 TP sc, respectively, during the 30-day treatment period. Blood was collected every 15 days and luteinizing hormone (LH), follicle-stimulating hormone (FSH) and testosterone were measured. Changes of weight and histological appearance of sex organs were determined at the end of the 30-day treatment and 15-day post-treatment periods. TP treatment reduced serum FSH and LH levels and significantly increased the weight of the seminal vesicles and epididymis, in accordance with histopathological changes, in both intact and orchidectomized rats. No changes in serum testosterone, LH, and FSH levels were observed in any of the intact rats treated with BS, but a significant increase in seminal vesicle weight was observed only in the BS-250 group. Although a significant reduction in serum LH was detected in the BS-50 and BS-250 groups of orchidectomized rats, no significant change in weight or histology of sex organs was observed. Thus, we conclude that B. superba needs endogenous testosterone to work synergistically to stimulate the accessory sex organ of intact animals and can potentially exhibit an LH reduction effect in orchidectomized animals.

Introduction

Butea superba Roxb. (Leguminosae: Fabales: Fabaceae), known in Thai as the red Kwao Krua, is a leguminous plant, which has been claimed to have aphrodisiac properties (1). Many products based on B. superba, such as a capsule formulation or gel cosmetic are claimed to support normal sexual function and to enhance sexual stamina, erectile capacity, sensitivity, and sexual performance, and are widely sold in local markets. It has also been reported that B. superba can improve erectile dysfunction in mature human males (2). It can increase intracavernous blood flow (3) and lead to erection via the inhibition of cGMP and cAMP phosphodiesterase activity (3-5). Accordingly, investigations have been carried out to evaluate the androgenic activity of B. superba on the reproductive system of male animals (6,7).

Manosroi et al. (6) treated intact male rats with a powdered suspension of B. superba at the doses of 2-1250 mg/kg body weight for 8 weeks and found that sperm counts increased by 16% relative to the control group, but without a dose-response relationship. Thus, they concluded that B. superba may contain compounds, which have androgenic activity and that these may increase the release of gonadotropin-releasing hormone (GnRH) from the hypothalamus, increase the release of male sex hormone and, in turn, stimulate the growth of Sertoli and Leydig cells. However, they did not measure the hormonal levels related to the hypothalamic-pituitary-testicular axis (6). In contrast, it has been recently reported that feeding B. superba at doses of 150 and 200 mg·kg body weight-1·day-1 for 90 days to intact male rats significantly reduced serum testosterone levels and slightly decreased serum luteinizing hormone (LH) levels, with a normal appearance of the testes observed under histological examination (7). The authors concluded that B. superba acts as an androgen disruptor, mainly through the alteration of testosterone biosynthesis or metabolism (7). Thus, an androgenic activity of B. superba in male animals has been suggested, but without strong experiments to support the conclusion. It has also been reported that B. superba at a dose of 250 mg·kg body weight-1·day-1 had an androgenic effect on female reproductive organs by increasing uterine thickness and the number of uterine glands in intact and ovariectomized rats (8).

On the basis of these considerations, we studied the androgenic activity of B. superba in intact and orchidectomized male rats by determining its effects on the pituitary-testicular axis and reproductive organs.

Material and Methods

Animals

Adult male Wistar rats aged 60 days and weighing 250-300 g were obtained from the National Laboratory Animal Center, Mahidol University, Nakhon Pathom, Thailand. They were housed in stainless steel cages with sawdust bedding at 5 animals/cage in a room with controlled lighting (lights on 6:00-20:00 h) and temperature (25 ± 1°C) at the Primate Research Unit, Department of Biology, Faculty of Science, Chulalongkorn University. The animals were fed rat chow (Pokaphan Animal Feed Co., Ltd., Thailand) and water ad libitum and were acclimatized to the surroundings for two weeks before starting the study. The experimental protocol was approved by the Animal Ethics Committee in accordance with the guide for the care and use of laboratory animals prepared by Chulalongkorn University.

Experimental procedure

Adult male rats used in this study were divided into two main groups: intact testes and orchidectomy. Each of these two main groups were randomly subdivided into five treatment groups (10 rats/group): distilled water (DW), Butea superba (BS)-10, BS-50, BS-250, and testosterone propionate (TP). DW, BS-10, BS-50, and BS-250 rats were gavaged with a suspension of 0, 10, 50, and 250 mg·kg body weight-1·day-1 B. superba in 0.7 mL distilled water, respectively, during the treatment period. In the TP group, rats were injected subcutaneously with 6 mg·kg body weight-1·day-1 TP in 0.2 mL sesame oil. The experimental schedule was divided into three periods: pre-treatment, treatment and post-treatment for 15, 30, and 15 days, respectively. Rats were administered distilled water during the pre- and post-treatment periods by gavage for the BS group and subcutaneous injection for the TP group.

Intact group. Blood samples (1 mL) were collected at 9:00-10:00 h on the first day and then every 15 days of the study period, designated as D1, D16, D31, D46, and D61. Immediately after collection, all blood samples were centrifuged at 1000 g at 4°C for 20 min, and sera were used for the determination of testosterone, LH, and follicle-stimulating hormone (FSH). At the end of the treatment period, half the rats (i.e., 5 rats from each group) were randomly euthanized with ether. The testes, epididymis and seminal vesicles were dissected, cleaned of connective and other tissues, weighed and then fixed in 10% (w/v) neutral buffered formalin solution and manipulated for histological examination as described previously (9). The remaining five rats in each group were euthanized at the end of the post-treatment period, and the testes, epididymis and seminal vesicles were processed as described. All rats were weighed once a week throughout the experimental period.

Orchidectomy group. Before submitting the rats to the pre-treatment period, a blood sample was collected at 9:00-10:00 h and the animals were then orchidectomized, and this day was designated as D-14. A 14-day recovery period was allowed before including the animals in the study. The experimental protocol for this group was similar to that described above for the intact group.

Preparation of B. superba suspensions

The tuberous roots of B. superba were collected from Lampang Province, Thailand (voucher specimen No. BCU 11046), as reported in Cherdshewasart et al. (10). The B. superba roots used throughout this study were from the same lot. The root was sliced and dried at 70-80°C, pulverized in a mortar, and filtered through a 100-μm mesh screen. The filtered powder was stored in an airtight container in the dark as a stock at room temperature. During treatment, the dried powder of B. superba was mixed with distilled water to obtain a stock suspension from which the required dilutions were made to a final volume of 0.7 mL of a final dose of 0, 10, 50, and 250 mg/kg body weight (8). The suspension was administered to the rats at 8:00-9:00 h using a gastric feeding needle.

Preparation of testosterone propionate

TP powder (Sigma, Merck, USA) was weighed and dissolved in a small volume of absolute ethanol (GR Grade, Merck, USA). After the powder had completely dissolved, sesame oil was added and the solution was allowed to stand at room temperature with the ethanol evaporated. This stock solution was then diluted with sesame oil to provide a final dose of 6 mg·kg body weight-1·day-1·200 µL sesame oil-1. The stock TP solution was kept in dark bottles at room temperature until used. The solution was injected subcutaneously into rats between 8:00 and 9:00 h.

Histological analysis

After overnight fixation in formalin, the testes, epididymis and seminal vesicles were dehydrated in a series of ethanol gradients, cleared in xylene and embedded and blocked in paraffin prior to preparing 5-µm sections and staining with hematoxylin and eosin, as reported (9,11). The permanent slides of testis, epididymis and seminal vesicle sections were then examined under an Olympus light microscope and representative sections were photographed.

The number of seminiferous tubules in intact male rats that contained a small number of spermatozoa, defined here as being <50% of the amount of spermatozoa observed in the normal seminiferous tubule of the control DW group, was evaluated histologically from the stained sections. Three sections/rat from each of 10 rats/group were counted.

Determination of serum testosterone, LH, and FSH levels

Serum testosterone levels were measured using the established radioimmunoassay technique of the World Health Organization (WHO) after the samples were extracted with ether (12).

Serum FSH and LH levels were measured by radioimmunoassay techniques using reagents obtained from the National Hormone and Pituitary Program. Iodination preparations were rat NIDDK-rat FSH-I-5 and rat LH-I-5. The antisera were anti-rat FSH-S11 and anti-rat LH-S11. The results obtained are reported in terms of the rat FSH-RP-2 and rat LH-RP-3 reference standards (13).

To minimize interassay variations, all samples were run in a single assay. The intra-assay coefficients of variation for testosterone, FSH, and LH were 10.6, 7.3, and 8.1%, respectively.

Statistical analysis

Data are reported as means ± SEM. The weights of the testis, epididymis and seminal vesicle collected at the end of the treatment and post-treatment periods were compared by the Student t-test. Differences in serum hormone levels between the pre-treatment, treatment and post-treatment periods in each group, and between the DW, BS, and TP groups in each experimental period, were analyzed by one- way analysis of variance (ANOVA) followed by the post hoc LSD test. In all cases significance was set at P < 0.05.

Results

Effect of B. superba on rats with intact testes

Serum testosterone, FSH, and LH levels. When compared to the pre-treatment period (D1), serum testosterone levels in rats from the negative control (DW) group did not change significantly throughout the study period (Figure 1A), while the testosterone levels of the positive control (TP- treated) group were markedly and significantly increased throughout the treatment period (D16-46) and then returned to the pre-treatment level during the post-treatment period (D61). Since the assay can detect the exogenous-injected TP, in addition to endogenous testosterone levels, this is to be expected and does not address by itself the endogenous levels. The average serum testosterone levels in the groups receiving the three B. superba dose (BS) were not significantly different from each other or from those of the control DW group throughout the study period.

The average serum FSH levels of rats from the control (DW) and all three B. superba dose (BS) groups were not significantly different from each other throughout the study period, both within each treatment group over time, and between treatment groups (all P > 0.05; Figure 1B). In contrast, serum FSH levels were markedly and significantly decreased (P < 0.01) in the TP group during the treatment period (D31-D46), but then returned to pre-treatment levels during the post-treatment period.

The changes in serum LH levels in all five groups of intact rats were broadly similar in pattern to those observed for the FSH levels (Figure 1C), with no significant changes within or between the DW control and all three BS treatment groups throughout the study period, but with a significant decrease in serum LH levels being observed in the TP group from D31 through D61 of the post-treatment period. In slight contrast was the weak recovery of serum LH levels in the TP-treated group during the post-treatment period.

Body weight and reproductive organ weight

Relative to D1, the mean rat body weights increased gradually over the 61-day period for the control (DW), TP and all three BS treatment groups, but there was no significant difference between them or compared to the DW control group at each time throughout the study period (data not shown). The body weight gain of the TP group was numerically lower than that of the DW and BS groups but there was no statistically significant difference between them.

There were no significant differences in average testis weights between the treatment and post-treatment periods in the DW and in each BS group, or between the control (DW) and all three BS groups (Figure 2A). In contrast, the average testis weight of the TP group during both the treatment and post-treatment periods was significantly lower than those of the control DW and BS treatment groups (P < 0.01) and, additionally, the testis weight of TP-treated rats during the post-treatment period was significantly lower than that observed during the treatment period (P < 0.05).

There were also no differences in average epididymis weight between the treatment and post-treatment periods in the control DW group and in each of the BS treatment groups, or between the control and all three BS treatment groups during the treatment and post-treatment periods (Figure 2B). In contrast, the average epididymis weight of the TP group during both the treatment and post-treatment periods was significantly higher than those of the DW and BS groups (P < 0.01) and, additionally, the weight during the post-treatment period was significantly lower than that during the treatment period (P < 0.05).

Changes in average seminal vesicle weight in the control and all three BS groups were similar to those of the epididymis weights, except that the seminal vesicle weight of the BS-250 group was higher than those of the control DW and the BS-10 and BS-50 treatment groups (P < 0.05; Figure 2C). In contrast, the average seminal vesicle weight of the TP group during both the treatment and post-treatment periods was significantly higher than those of the DW and BS treatment groups (P < 0.01), while the weight during the post-treatment period was significantly lower than that during the treatment period (P < 0.05).

Histology of testis, epididymis and seminal vesicle

The histology of the testis in DW and BS-treated rats during the treatment period showed numerous spermatogenic cells in various stages, including primary spermatocytes (S1), secondary spermatocytes (S2), spermatids (S3), and spermatozoa (S) (Figure 3). In contrast, the testis of TP-treated rats showed a thin layer of spermatogenic cells and a small number of spermatozoa when compared to the control DW and all three BS-treated groups. The testis histology for the DW and BS groups did not differ between the treatment and post-treatment periods, but spermatogenic cell types seemed to increase in the TP group during the post-treatment period. Thus, the percent of seminiferous tubules with a low sperm number counted (or impaired spermatogenesis) did not differ between the DW and the three BS-treated groups, but was significantly higher (P < 0.01) in the TP group, which was in a partially recovered state during the post-treatment period (Table 1).

During the treatment period, the histology of the epididymis of control DW rats and of rats treated with all three doses of BS showed a similar composition mainly consisting of columnar and cuboidal ciliated epithelial cells (EP) with numerous spermatozoa (S) inside the tubules (Figure 3). Comparable to the reduction of sperm production in the testis, the sperm number in the epidymidis of TP-treated rats was also decreased, and the columnar ciliated cells showed a thicker layer. The histology of the epididymis of the control DW group and of all three BS-treated groups did not differ between the treatment and post-treatment periods. However, during the post-treatment period the epididymis of the TP group showed a thinner layer of epithelial cells.

The seminal vesicles of control DW rats and of those treated with all three doses of BS showed a high papilla folding (EX) of the tubular glands and muscular layers (Figure 3). A whitish-yellow viscous material (SF) was secreted into the lumen of the seminal vesicle (Figure 3). In the BS-250 group, the folded tubular glands were similar to those of other BS groups, but the amount of secretory material was higher. In the TP group, the number of folded tubular glands and the level of secretory material were higher than those of the DW and BS groups. While there were no differences in seminal vesicle histology between the treatment and post-treatment periods in the DW and BS groups, in the TP group the number of folded tubular glands and the quantity of secretion material were lower during the post-treatment period than during the treatment period (Figure 3).

Effects of B. superba in orchidectomized rats

Serum testosterone, FSH, and LH levels. There were no significant differences between the five groups before (D-14) and after (D1) orchidectomy. However, for 14 days after orchidectomy, serum testosterone levels of rats were significantly decreased (286.1 ± 56.9 and 17.7 ± 2.1 pg/mL for D-14 and D1, respectively; Figure 4A). Compared to D1, there were no significant changes in serum testosterone levels in the DW and BS groups throughout the study period, whereas the serum testosterone levels of TP-treated rats were significantly increased (P < 0.01) during the treatment period to over a 3-fold higher level than that prior to orchidectomy. Although serum testosterone levels decreased during the post-treatment period, and fell back below the pre-orchidectomy level, they were still significantly higher than the D1 level (P < 0.05).

In agreement with the decrease in serum testosterone levels, serum FSH levels of rats were significantly increased by 14 days after orchidectomy (5.3 ± 0.3 and 26.4 ± 2.9 ng/mL for D-14 and D1, respectively; Figure 4B), and compared to the D1 levels, continued to increase significantly throughout the study period in the DW and BS groups. Although the serum FSH levels in all three BS-treated groups were numerically lower than those of the DW group, there was no statistically significant difference. In contrast, during treatment, the serum FSH levels of TP-treated rats were significantly lower than those of the control DW group (D31-D46), before rising again during the post-treatment period.

Likewise, in agreement with the changes in serum testosterone and FSH levels, serum LH levels of rats were significantly increased by 14 days after orchidectomy (0.7 ± 0.2 and 21.4 ± 3.3 ng/mL for D-14 and D1, respectively; Figure 4C). Serum LH levels of control DW rats (P < 0.01) and of the BS-10 treatment group (P < 0.05) increased significantly throughout the study period but no statistically significant differences were observed between these two groups. In contrast, the serum LH levels of rats from the BS-50 and BS-250 treatments were significantly lower than those of the control DW group at D31-D61 (P < 0.01). The serum LH levels of TP-treated rats started to be lower than those of the DW group from D16, and were significantly different on D31-D61 (P < 0.01).

Body weight and reproductive organ weight

Relative to D-14, the mean rat body weights gradually increased numerically over the 75-day period for the control DW, TP, and all three BS treatment groups, but with no significant difference between each other or compared to the DW group at each time throughout the study period (data not shown). Thus, at least under these unrestricted food and standardized conditions, the BS treatments had no effect on body weight. In contrast, the body weight gain of TP-treated rats at the end of the study period was significantly higher than those of the control DW and all three BS treatment groups (124.5 ± 10.8 g vs 70.2 ± 7.9 g, respectively) but there were no differences between DW, BS10, BS50, and BS250 groups.

There were no significant differences in the average epididymis and seminal vesicle weights between the treatment and post-treatment periods within the control DW and each of the three BS treatment groups, or between the control DW and all three BS treatment groups (Figure 5A,B). In contrast, the average epididymis and seminal vesicle weights of TP-treated rats during both the treatment and post-treatment periods were significantly higher than those of the DW and BS groups (P < 0.01), and the weights during the post-treatment period were significantly lower than during the treatment period (P < 0.05).

Histology of the epididymis and seminal vesicle

Orchidectomized rats showed the absence of stereocilia in the ductus tubulus epididymis, a smaller lumen size with many layers of vacuoles in the epithelial lining (EP) and the absence of spermatozoa in the lumen during the treatment period compared to intact male rats (Figure 6). There were no differences in the histology of the epididymis between the treatment and post-treatment periods in the control DW or in each of the three BS doses, nor between the control DW and all three BS groups (Figure 6). In contrast, during the treatment period the tubular epithelium of the ductus epididymis of the TP group was pseudostratified, consisting of tall columnar principal cells with long sterocilia and small basal cells without vacuoles. During the post-treatment period, the number of stereocilia and their height in the tubular epithelial cells were diminished.

After orchidectomy the DW control and BS-treated rats showed a decrease in the number of epithelial foldings (EX) and an absence of seminal secretion (SF; Figure 6). In contrast, TP-treated rats revealed a highly developed papilla folding pattern of the seminal vesicle tubular glands with numerous primary, secondary, and tertiary foldings, filled with secretory material, which then decreased during the post-treatment period.

Discussion

Although the products prepared from B. superba are widely consumed for various reproduction-related activities in men, the androgenic activity of B. superba is not known. The present study was, therefore, carried out to determine the androgenic effects of B. superba on the pituitary-testis axis and the reproductive organs (weights and histology) of intact and orchidectomized adult male rats. Distilled water and TP (6 mg·kg body weight-1·day-1) were used as negative and positive controls of androgenic activity, respectively (8,14-17). The doses of B. superba used in this study, 10, 50, and 250 mg/kg body weight, were based on previous studies carried out with female (8) and male (6,7) rats.

The response of body weight gain of adult male rats to TP treatment was decreased in intact and increased in orchidectomized rats, despite the fact that orchidectomy decreased the body weight gain in the DW control group. These changes are in accordance with previous reports (14-16), but are different from the reported effects of estrogens and ovariectomy (8,18,19).

The positive control (TP-treated group) showed the expected androgenic effects occurring in intact male rats that is decreased serum LH and FSH levels. Indeed, after FSH and LH secretion from the pituitary gland is reduced (20), the endogenous testosterone secretion from Leydig cells should subsequently be reduced. Although the serum LH and FSH levels were decreased during the TP treatment, the extent of reduction of LH levels was greater than that of FSH levels, and only the serum FSH levels returned to pre-treatment values during the post-treatment period. These differences are expected to be largely attributable to the effect of inhibin, the main suppressor of FSH secretion in rats (21). The decrease in serum LH and FSH levels induced by the TP treatment causes a decline in testosterone production by Leydig cells and, therefore, a reduction in the intratesticular testosterone concentration that, in turn, causes a shrunken testis and reduces sperm production in the seminiferous tubules of intact male rats (17). This likely explains the decreased testis weight that was detected in the TP-treated rats in the present study. In contrast to the decline in testicular weight with TP treatment, the seminal vesicle and epididymal weights of the rats increased and the hypertrophy corresponded to mild, though noticeable, histological changes in cell growth and secretion (14,16,17,22).

In contrast to the effects of TP, B. superba powder did not alter the serum testosterone, LH, and FSH levels or the weights and histological appearances of the reproductive organs (testis and epididymis) in intact male rats, except that the seminal vesicle weight increased only in the BS-250 group. It was previously reported that B. superba significantly reduced serum testosterone levels with no abnormal appearance of the testicular and epididymal histology or tissue weights. However, this study did not determine if there were any changes in the seminal vesicle (7), the most sensitive organ of androgenic or anti-androgenic activity in the Hershberger bioassay (16,22).

Complete orchidectomy caused lower serum testosterone and higher serum FSH and LH levels in orchidectomized rats than in normal male rats. No effects of B. superba on the serum testosterone levels of orchidectomized rats were observed in the present study, although BS-50 and BS-250 significantly suppressed the increased serum LH levels. In contrast, serum LH and FSH levels were significantly decreased in TP-treated rats after 15 days of treatment. Thus, we conclude that the doses of B. superba administered can partially suppress the hypothalamic-pituitary axis in orchidectomized rats, which seems to indicate the weak androgenic activity of B. superba. At present, it is unknown what chemicals in B. superba exhibit androgenic activity and further investigation is still needed. B. superba root contains the flavonoid and flavonoid glycoside (4,23), which showed inhibitory effects on cAMP phosphodiesterase activity (4). Currently, three phytoestrogens, daidzein, coumestrol and genistein, have been isolated from the tuberous root of B. superba (24). Genistein reduced the pituitary contents and prostate weights of male mice (25), interfered with the coupling of transmembrane LH receptors to G proteins and suppressed the steroidogenesis of the testicular Leydig cells in adult male rats (26). Coumestrol can suppress the pulsatile LH secretion from the pituitary gland of ovariectomized rats (27). Previously, we reported that Pueraria mirifica, a phytoestrogen-containing herb, can suppress serum LH and FSH levels in female as well as in male rats, although the response of females was greater than that of males (13). In addition, orchidectomized rats are more sensitive to weak endocrine disruptors than intact rats (28). Taken together with no changes in weight or histological appearances of the epididymis and seminal vesicles in all three BS treatment groups of the orchidectomized rats, this suggests that the decrease in serum LH levels in BS-50- and BS-250-treated rats is caused by a weak estrogenic activity of phytoestrogen constituents in B. superba. It is difficult to know if the reduction of LH levels in orchidectomized rats is due to the estrogenic activity or androgenic activity of B. superba. Higher doses of B. superba are suggested to be used. However, because the B. superba powder suspension has a high viscosity, a dose higher than 250 mg/kg body weight, the highest dose used in the present study, could not be administered by gavage. Thus, each substance isolated from B. superba should be tested separately for androgenic activity as well as estrogenic activity in male rats.

On the basis of the results obtained here in intact and orchidectomized rats, we conclude that B. superba needs to work synergistically with an endogenous testosterone to stimulate accessory sex organ in intact animals and can potentially exhibit an LH reduction effect in orchidectomized animals.

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Anaheim, California: ‘They tortured and locked me up for four years’, Kerala domestic help reveals torture saga

David B. Bailey 3692 Elk City Road Indianapolis, IN 46250

The 20-year-old Nepali speaking girl said she was thrashed for small mistakes by her employer. In the Malayalam movie Gaddama, Kavya Madhavan played the role of a domestic help trapped and tortured in an Arab's house in the Middle East. The character’s agony portrayed the life of many Malayalis who go abroad to earn a living, working in harsh conditions for a pittance.

But the reality is that the plight of some domestic helps is no different in Kerala.

On May 1, the police rescued a 20-year-old Nepali speaking woman from Vypeen in Ernakulam. Sarsu Halsan, who hails from Jalpaiguri, West Bengal was tracked down by the police based on a missing complaint filed by her employer on April 29.

“I had no parents, both of them died and I lived with my aunt. A distant cousin of mine brought me to Kochi when I was 16-year-old. I joined as a domestic help at this house,” Sarsu told The News Minute, outside Kalamassery police station.

Sarsu was working at the house of Kavita Nair*, in Kochi. “Before I joined she gave me Rs 3,500 and that was the only amount I received in the last four years. Initially she had told my cousin that I would be paid Rs 5,000 monthly. But I did not get any money,” Sarsu said.

But what was more intolerable for her was the physical torture she had to go through. “For even small mistakes I used to get thrashed. She hit me using sticks, hands, broom or some large spoons. She also banged me against the wall. I used to always end up bleeding,” Sarsu said, showing the scars on her hands.

“I ran away from the house to commit suicide. I did not want to live,” she said.

As Sarsu is led away by workers of a rescue home, she screams out, “She cut my hair forcefully, never allowed me to make calls or write a letter,”

What Sarsu described was only a small part of what she actually went through in the past four years. “There are scars from wounds all over her body, covered in blood clots. The torture was quite severe,” E V Shibu, Station House Officer at Kalamassery, said.

The police officer said that Sarsu had been trying to escape from the house for the past four years but was not successful because she was always locked up.

“On April 29 she managed to lock up her employer and her daughter inside a room and ran away from the house. She went to a beach near Njarackal hoping to die. She stayed in a house nearby, and somebody informed us about her. We went and picked her from there,” the officer said.

Kavita claims that she had paid Sarsu’s salary to a relative of hers in Jalpaiguri. Sarsu was brought to work as a domestic help when she was still a minor, which means that the employer can charged under the Juvenile Justice Act. Kavita can also be charged under the act for harassment.

“We should confirm the date tongkat ali dosage when the girl was brought in for the work and also need to get more evidence before registering the arrest,” the police officer said.

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Fresno, California, USA news

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Feminist rule in Europe makes second-generation male Muslim immigrants suicide bombers. They die for sexual justice. Why do Western politicians call suicide bombers cowards? To sacrifice one's own life is the ultimate in courage.

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Butea superba conditions the mind for superb sex. And don't underestimate the power of the mind. If your mind is in tune for optimal sex, you will reach 100 years and still enjoy doing it.

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Corpus Christi, Texas: Testosterone linked to entrepreneurial ambition

Joshua M. Williams 4803 Davis Avenue Hydesville, CA 95547

THE desire to work for oneself is linked to higher testosterone levels, a research study at the Warwick Business School in the UK concluded.

Nicos Nicolaou, Professor and Head of the Entrepreneurship and Innovation Group at Warwick Business School said that high levels of testosterone can give individuals the push they need to work for themselves.

“Using the most widely accepted methods available for measuring testosterone levels and analyzing three diverse samples, our findings indicate testosterone levels may constitute an important influence on the likelihood individuals will engage in self-employment,” Nicolaou explained in an email.

“The study also utilized a new research design involving opposite-sex and same-sex twins to contribute to the ongoing debate regarding the significance and validity of the relationship between testosterone and self-employment,” he added.

Nicolaou said the research was inspired by the ongoing debate over whether business behaviors are learned or can be at least partly attributed to biology.

“Our research shows it is indeed possible that at least a portion of certain business behaviors can at least in part be attributed to biological influences,” he said. “Our results represent an important first step into uncovering how key biological influences are related to self-employment and entrepreneurial activities.”

Three separate studies were conducted as part of the research. In the first, data from the Centers for Disease Control and Prevention’s National Health and Nutrition Examination Surveys (NHANES) of 2011-2012 was analyzed, and found 2,146 observations that suggested a link between higher levels of testosterone and self-employment in males.

In the second study, Nicolaou examined the 2D:4D digit ratio—the ratio of the length of the index finger to the length of the ring finger, which is a common marker of prenatal testosterone exposure —to determine if there was a correlation to self-employment, surveying 449 males and 525 females.

The results indicated males with a lower 2D:4D ratio in their left hand, or higher prenatal testosterone exposure, have a significantly greater likelihood of self-employment. This was also found to be marginally significant for females.

The third study examined the twin testosterone transfer effect in a sample of opposite-sex and same-sex twins from the National Survey of Midlife Development in the US.

Nicolaou explained that previous studies have suggested that female fetuses gestated with a male twin are more likely than female fetuses gestated with a female twin to be “masculinized” in their development and to have greater testosterone levels. This is because testosterone may pass from one twin to the other through maternal circulation and by diffusion through fetal membranes.

Professor Nicolaou found that these females were more likely to be self-employed than females gestated with a female twin.

“The findings are relevant to both entrepreneurship and management audiences,” Nicolaou wrote in the research report, “Testosterone and Tendency to Engage in Self-employment,” which is to be published in an upcoming issue of Management Science.

“Higher levels of testosterone can not only enhance an individual’s willingness to take risks but also diminish the likelihood that they feel fear with regards to risky situations, when coupled together it is possible that individuals with higher levels of testosterone could be prone to engage in entrepreneurial activities and self-employment,” he explained.

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The Breast Ripper was often used heated during torture and it contained four claws, which were used to slowly rip the breasts from women for various crimes. The instrument would be imposed onto a single breast of the woman. They were designed to shred, or tear off the breasts of the victim. If the woman did not die, she would be horribly disfigured for the rest of her life.

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Lexington-Fayette, Kentucky: After long wait, Japan moves to ban possession of child pornography

Michael A. Rogers 4475 West Drive Chicago, IL 60606

In most of Japan, it's still legal to possess child pornography. Although production and distribution have been banned for 15 years, Japan lags behind other major developed nations in forbidding people from simply holding the sinister material.

That is about to change in a country regarded as a global nexus of child pornography. The country's upper house of parliament is expected to pass legislation this month making possession of it a crime punishable by up to a year in prison. Children's rights activists have applauded the step, although their reaction is tempered with frustration that it has taken such a long time.

"As a member of a group that's been hearing the voice of the victims for many years, we welcome the news," said Shihoko Fujiwara, a representative of Lighthouse, a nonprofit group that helps exploited children. "Japan took so long, and it is too late to reach this decision as a developed country."

The proposed law, which was already approved by the lower house of parliament this week, comes with a couple of noteworthy loopholes. When it goes into effect, it will give those already in possession of child pornography a year to dispose of it.

And it won't cover the country's popular manga (comic book) and anime (animation) industries, which include depictions of violent sexual abuse of children in their publications.

Fujiwara said a discussion about some of the imagery in manga and anime -- content that would be illegal in many Western countries -- would be a natural "next step."

'A necessary evil'

But representatives of those industries say that while they support the ban on real child pornography, any move to censor their products would be an unjustified restriction of freedom of expression. Daisuke Okeda, a lawyer and inspector for the Japan Animation Creators Association, said it was "natural that animation is exempted."

"The goal of the law itself is to protect children from crime," he said. "Banning such expression in animation under this law would not satisfy the goal of the law."

Okeda said that no studies have been done that prove any link between pedophilia and animation in Japan.

Hiroshi Chiba, the manager of Chiba Tetsuya Production, one of the country's best known manga production houses, said that more could be done in terms of age restrictions on graphic content featuring children and to distinguish it more clearly from other comics. And he admitted that some products of the industry leave him and his colleagues "disgusted."

"But rich, deep culture is born from something that might not be accepted by all," Chiba said. "We need to allow the gray zone to exist as a necessary evil."

'An international hub'

Some experts counter that children suffer in a culture that appears to tolerate images of child sexual abuse.

Hiromasa Nakai, a public affairs officer for UNICEF in Japan, pointed to the graphic content in manga, anime and some video games, as well as the "junior idol" genre of books and DVDs that display minors wearing tiny bikinis and striking sexual poses.

Japan should do more -- beyond the proposed law change -- "to protect the best interest of children," Nakai said.

Statistics show that child pornography remains a big problem in Japan.

The U.S. State Department's 2013 report on human rights practices in Japan labels the country "an international hub for the production and trafficking of child pornography."

It cited Japanese police data showing the number of child pornography investigations in 2012 rose 9.7% from a year earlier to a record of 1,596. The cases involved 1,264 child victims, almost twice as many as in the previous year.

The fact that possession remains legal, for the time being, "continued to hamper police efforts to enforce the law effectively and participate fully in international law enforcement," the report said.

Girls as sex objects

One local authority already took matters into its own hands. The prefecture of Kyoto in central Japan introduced a ban on possession of child pornography in 2011.

But Nakai said addressing the problems isn't just a matter for government, suggesting parents, the media, the private sector and even children themselves can play a role in improving the situation. The portrayal of young girls as sex objects in Japan has long raised eyebrows among Westerners.

An article in Wired in 1999 reeled off a list of examples in Tokyo: "Vending machines sell schoolgirls' used panties, which the girls sell to middlemen. 'Image bars' specialize in escorts dressed in school uniforms. Telephone clubs feature bored adolescent girls earning spending money by talking dirty. Sex shops sell a porn magazine called 'Anatomical Illustrations of Junior High School Girls.'"

Some experts suggest the situation is born out of Japan's long-established patriarchal society.

Whatever the cause, changing a culture may prove a lot harder than changing a law.

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The world in 200 years will be populated by a few thousand male humans who live indefinitely, and a huge number of female looking robots. Women aren't needed, really, and anyway, women are troublemakers, more than anything else.

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Arthur Schopenhauer, the greatest German philosopher, on women: Only a male intellect clouded by the sexual drive could call the stunted, narrow-shouldered, broad-hipped and short-legged sex the fair sex … More fittingly than the fair sex, women could be called the unaesthetic sex. Neither for music, nor poetry, nor the plastic arts do they possess any real feeling of receptivity: if they affect to do so, it is merely mimicry in service of their effort to please.

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Cincinnati, Ohio: The protective effect of Mucuna pruriens seeds against snake venom poisoning

Victor S. Deal 4722 Goldie Lane Cincinnati, OH 45202

Ethnopharmacological relevance

The seed, leaf and root of Mucuna pruriens have been used in traditional medicine for treatments of various diseases. In Nigeria, the seed is used as oral prophylactics for snakebite.

Aim of the study

To study the protective effects of Mucuna pruriens seed extract against the lethalities of various snake venoms.

Materials and methods

Rats were pre-treated with Mucuna pruriens seed extract and challenged with various snake venoms. The effectiveness of anti-Mucuna pruriens (anti-MPE) antibody to neutralize the lethalities of snake venoms was investigated by in vitro neutralization.

Results

In rats, MPE pre-treatment conferred effective protection against lethality of Naja sputatrix venom and moderate protection against Calloselasma rhodostoma venom. Indirect ELISA and immunoblotting studies showed that there were extensive cross-reactions between anti-MPE IgG and venoms from many different genera of poisonous snakes, suggesting the involvement of immunological neutralization in the protective effect of MPE pre-treatment against snake venom poisoning. In vitro neutralization experiments showed that the anti-MPE antibodies effectively neutralized the lethalities of Asiatic cobra (Naja) venoms, but were not very effective against other venoms tested.

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St. Louis, Missouri: Not so cool now! Pro-rape pick-up artist pictured in a sweat-stained T-shirt at the door of his mother's home (where he lives in the BASEMENT!)

Marcus L. Aliff 1687 Mutton Town Road Seattle, WA 98101

This is the man at the center of a worldwide storm after advocating legalizing rape on private property - in a sweat-stained T-shirt at the door of his mother's house.

Daryush 'Roosh' Valizadeh, 36, the self-proclaimed 'King of Masculinity' called police after receiving death threats from around the world and canceled a series of 'tribal meetings' in 45 countries set for this weekend.

Valizadeh, who is at the center of public protests at home and in Canada, Australia and the UK, is on record as advocating women be banned from voting, describing a woman's value as dependent on her 'fertility and beauty', and stating that women with eating disorders make the best girlfriends.

In a highly-criticized blog he said that if a woman was raped on private property, it should be legal.

Today he told police that it was meant to be a satirical article and that he had written it in early 2015 and had since put a disclaimer on the piece saying it was satire.

But asked when he had added the disclaimer he admitted it had been placed only 'yesterday'.

The internet geek, who has written a series of books teaching what he claims is the best way for men to use their testosterone to bed women, likes to portray himself as an global businessman.

But as the international storm grew around him today, Daily Mail Online found him in hiding at the cul-de-sac where he ekes out his vile views on his laptop - and sells ads on his website, which cost $150 a day.

Today, dressed in a stained T-shirt and shorts and living in the basement of his mother's home, he was concerned for his safety.

He said he had received death threats from around the world. He played officers voicemails left on his phone and showed them emails.

Some were from Britain, Australia and the US and warned him he would be 'shot, stabbed or have his home burned down.'

One said: 'We will kill you if you come to our city' and others were filled with vitriol, he told officers.

After dialing 911, two officers visited him and he greeted them in his work attire. One female officer only entered his doorway and he had to bring his laptop to the stoop and front hall to show her how his views on rape had backfired.

He said he had only been aiming to gain attention but had not budgeted for the worldwide anger against him and feared for his safety.

Valizadeh, who used the alias Roosh, said he was canceling the worldwide city weekend meetings of his followers after the threats.

He said he could 'no longer guarantee the safety or privacy of the men who want to attend'.

In a statement he posted online, he apologized to his supporters and said they would be let down.

Meetings had been planned around the US including Washington, New York and Los Angeles and across the globe.

Valizadeh had said he would be attending a gathering in Australia, but backed down after a public outcry there which was echoed, particularly in Britain where 80,000 signed a petition calling on the government to ban him and his meetings using hate crime laws.

He had banned homosexual men from attending as well as all women

If a pretty girl approached a man attending, his advice online to followers was 'Get her number and then tell her to buzz off. Do not allow women to attend the meeting.'

He had advised followers that feminists may attack them or male opponents, but they were not to strike back but follow the 'Gandhi principle of non violence' record incidents on cell phones.

He graduated from the University of Maryland with a degree in microbiology and soon after started a local blog called DC Bachelor.

His first book called Bang was 'a textbook for picking up girls and getting laid.' He wrote several other books with the word 'Bang' in the title such as 'Day Bang'.

In America, he was placed on a 'misogyny list' by the social justice organization Southern Poverty Law Center.

Valizadeh celebrates and dwells on the title given to him when he visited Romania of 'World Don Juan.'

He says: 'I didn't try to become infamous worldwide, but that has been the outcome, all because of my teachings and ideas.

'I've been falsely accused of crimes like rape and harassment by my enemies in an attempt to shut me down, but they are too weak to defeat me.'

He has also complained about the abuse he has received over his views. One message was directed at Glasgow, Scotland, where he claimed 'I've received more threats from Glasgow than anywhere else combined. Is it some kind of convict resettlement zone?'

One Twitter user who replied was comedian and BBC broadcaster Hardeep Singh Kohli, who said: 'It's a city with a moral compass and a degree of self esteem. Try it sometime...'

Singh Kohli was suspended from his working with the broadcasters The One Show in 2009 over alleged 'inappropriate sexual behavior' towards a female colleague. No formal complaint was made and he apologized unreservedly for his behavior and later said: 'We all make mistakes and we all make misjudgments.'

A neighbor of the self proclaimed lothario said she was disgusted at his views.

Esther Eyere,33, a nursing student at Marymount University, said: 'I can't believe he can have views like that, especially about rape. It makes me sick.'

The U.K. government has called for him to be 'ridiculed' and welcomed the cancellation of his meetings.

Britain's Home Office Minister Karen Bradley told parliament today: 'The government condemns in the strongest terms anyone who condones rape and sexual violence.

'We should ridicule, we should show contempt, and we should show that these are the most ridiculous views.

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Feminism in Europe makes second-generation male Muslim immigrants suicide bombers. Only the patriarchy as a social and political system can achieve justice.

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It is only a question of time until butea superba will be outlawed in the Western World. In some people, it can cause hypersexualization that can last for weeks. And it can easily be added to food to improve taste. Imagine a Thai restaurant breeding hundreds of super horney women prowling for any man they can get, and that for weeks on end

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Restaurants in New York City, USA

The Odeon
145 W Broadway
New York, NY
(212) 233-0507

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Mission Chinese Food
171 E Broadway
New York, NY
(212) 432-0300

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Spicy Village
68B Forsyth St
New York, NY
(212) 625-8299

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Le Coucou
138 Lafayette St
New York, NY
(212) 271-4252

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Wildair
142 Orchard St
New York, NY
(646) 964-5624

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Uncle Boons
7 Spring St
New York, NY
(646) 370-6650

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Balthazar
80 Spring St
New York, NY
(212) 965-1414

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Lure Fishbar
142 Mercer St
New York, NY
(212) 431-7676

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Katz's Delicatessen
205 E Houston St
New York, NY
(800) 446-8364

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Somtum Der
85 Avenue A
New York, NY
(212) 260-8570

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Oiji
119 1st Ave
New York, NY
(646) 767-9050

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I Sodi
105 Christopher St
New York, NY
(212) 414-5774

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Cosme
35 E 21st St
New York, NY
(212) 913-9659

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