Deuterium labeling improves the therapeutic index of 3,3'-diselenodipropionic acid as an anticancer agent: insights from redox reactions.

3,3'-Diselenodipropionic acid (DSePA), a selenocystine derivative, has been previously reported as an oral supplement for anticancer/radio-modulation activities. The present study is focused on devising a strategy to synthesize and characterize the deuterated derivative of DSePA and on understanding the effect of deuteration on its therapeutic index by comparing its cytotoxicity in cancerous versus non-cancerous cell types. In this context, the synthesis of 3,3'-diselenodipropionic acid-D8 (D-DSePA) was accomplished in ∼42% yield. Further, the results clearly established that the deuteration of DSePA significantly reduced its cytotoxicity in non-cancerous cell types while retaining its cytotoxicity in cancerous cell lines. Together, D-DSePA displayed a ∼5-fold higher therapeutic index than the non-deuterated derivative for anticancer activity. The biochemical and NMR studies confirmed that the better biocompatibility of D-DSePA than its non-deuterated derivative in non-cancerous cells was due to its ability to undergo slower redox reactions and to cause lesser inhibition of intracellular redox enzymes.

Interaction of a Model Hydrophobic Drug Dimethylcurcumin with Albumin Nanoparticles.

The aim of present study was to investigate the binding interactions of a model hydrophobic molecule, dimethylcurcumin (DMC) with nanoparticle form of bovine serum albumin (BSA) using fluorescence spectroscopy techniques. For this, BSA nanoparticles (size = 62.0 ± 3.5 nm, molecular weight = 11,243 ± 3445 kD) prepared by thermal denaturation method was mixed with DMC in solution and monitored for fluorescence emission of tryptophan (Trp) residue as well as DMC separately. The emission maximum of DMC in nanoparticles form exhibited more blue sift and quenched the excited state of tryptophan (Trp) by six fold higher than in the native form of BSA. By analyzing Trp fluorescence, the mean binding constant (K) estimated for the interaction of DMC with native and nanoparticles forms of BSA was 2.7 ± 0.4 × 104 M-1 and 1.5 ± 0.5 × 105 M-1 respectively. Together these results suggested that DMC experienced a more rigid environment in nanoparticles than in native form of BSA. Additionally the above determined K values were in agreement with those reported previously by absorption techniques. Further direct energy transfer was observed between Trp and DMC, using which the distance (r) calculated between them was 28.25 ± 0.27 Çº in BSA native. Similar analysis involving BSA nanoparticle and DMC revealed a distance of 24.25 ± 1.05 Çº between the hydrophobic core and the ligand. Finally interaction of DMC with BSA was validated through molecular docking studies, which indicated sub-domain IIA as the binding site of DMC. Thus it is concluded that intrinsic fluorescence of protein can be utilized to study the interaction of its different physical forms with any hydrophobic ligand.

Tuning the binding, release and cytotoxicity of hydrophobic drug by Bovine Serum Albumin nanoparticles: Influence of particle size.

To elucidate the effect of particle size of albumin nanoparticles on cellular uptake of a hydrophobic drug, herein we report the release kinetics and cytotoxicity of nanoparticle bound dimethylcurcumin (DMC) in A549 tumor cells. The bovine serum albumin (BSA) nanoparticles were prepared by thermal denaturation and characterized by dynamic light scattering (DLS), zeta (ζ) -potential, circular dichroism (CD) and transmission electron microscope (TEM). The preparation conditions were optimized to obtain nanoparticles with mean hydrodynamic diameters 28.0nm (BSAnp1) and 52.0nm (BSAnp2) and corresponding ζ- potential value of∼-7.0 and -6.0mV, respectively. Interaction of DMC with BSA nanoparticles was investigated by UV-vis, fluorescence and CD spectroscopy. CD studies indicated significant changes in the secondary structure of BSA upon particle formation, as revealed by decrease in the helicity. The cellular uptake of DMC increased with increase in particle size and the toxicity of DMC loaded nanoparticles to A549 cells were found to be consistent with their cellular uptake. Between the two formulations studied, BSAnp2 provided enhanced cellular uptake and can be used as an effective delivery system for hydrophobic drugs like DMC.

Removal of As(V) by Cu(II)-, Ni(II)-, or Co(II)-doped goethite samples.

The present study reports removal of As(V) by adsorption onto laboratory-prepared pure and Cu(II)-, Ni(II)-, and Co(II)-doped goethite samples. The X-ray diffraction patterns showed only goethite as the crystalline phase. Doping of ions in the goethite matrix resulted in shift of d-values. Various parameters chosen for adsorption were nature of adsorbent, percentage of doped cations in goethite matrix, contact time, solution pH, and percentage of adsorbate. It was observed that the pH(pzc) of the goethite surface depended on the nature and concentration of metal ions. The surface area as well as the loading capacity increased with the increase of dopant percentage in goethite matrix. A maximum loading capacity of 19.55 mg/g was observed for 2.7% Cu(II)-doped goethite. The adsorption kinetics for Ni(II), Co(II) and for undoped goethite attained a quasi-equilibrium state after 30 min with almost negligible adsorption beyond this time. In case of Cu(II)-doped goethite samples, the quasi-equilibrium state for As(V) adsorption was observed after 60 min. At each studied pH condition, it was observed that the percentage of adsorption of As(V) decreased in the order Cu(II)-doped goethite > or = Ni(II)-doped goethite > Co(II)-doped goethite > pure goethite. The adsorption followed: Langmuir isotherm, indicating monolayer formation.

Effect of short-term cyclic administration of cyproterone acetate on pituitary-ovarian function in the human.

Short courses of cyproterone acetate, a compound with progestational and antiandrogenic activities, were administered to normally menstruating women during different phases of the menstrual cycle to suppress growth and maturation of the follicles and corpus luteum function. Postovulatory administration of 20 mg of the drug daily for 8 days to two women delayed menstruation by 4 to 6 days, followed by prolonged bleeding and short post-treatment cycles. Plasma levels of progesterone were suppressed temporarily during therapy, but increased immediately after cessation of treatment. Administration of 10 mg of the drug for 8 days during the early follicular phase to two women resulted in irregular bleeding, short cycles, and decreased plasma levels of progesterone throughout the cycle. Reduction of the dose to 2.5 mg during the early follicular phase in two other women also resulted in irregular cycles. When the 2.5-mg dose was administered to three women from the 8th to the 15th days of the cycle, vaginal bleeding and cycle length were normal. Plasma levels of luteinizing hormone and progesterone were suppressed during therapy. In one subject, cervical mucus was found to be hostile to sperm penetration in all three treatment cycles. The results indicate that, with cyclic administration of low doses of cyproterone acetate to women during the late follicular phase, it may be possible to interrupt pituitary-ovarian function, as well as sperm transport through the cervical mucus.

PIP: The effect of short-term cyclic administration of cyproterone acetate (CPA) on pituitary-ovarian function was investigated in the human. Postovulatory administration of 20 mg CPA/day for 8 days to 2 normally menstrauting women delayed menstraution by 4-6 days, followed by prolonged bleeding and short posttreatment cycles. Plasma progesterone levels were suppressed temporarily during therapy but increased immediately after cessation of treatment. Administration of 10 mg CPA for 8 days during the early follicular phase to 2 women resulted in irregular bleeding, short cycles, and decreased progesterone levels throughout the cycle. A 2.5 mg dose of CPA during the early follicular phase in 2 other women also resulted in irregular cycles. When the 2.5 mg dose was administered to 3 women from the 8th to 15th days of the cycle, vaginal bleeding and cycle length were normal. Plasma progesterone and luteinizing hormone were suppressed during therapy. In 1 woman cervical mucus was found to be hostile to sperm penetration in all 3 treatment cycles. These data indicate that with cyclic administration of low doses of CPA to women during the late follicular phase, it may be possible to interrupt pituitary-ovarian function as well as sperm transport through the cervical mucus.

The binding patterns of antisera to sex steroids and human gonadotropins on human and rhesus monkey spermatozoa.

The presence of different hormones on the surface of ejaculated spermatozoa was determined by immunofluorescence studies of the binding patterns of specific antisera to these hormones. There were striking similarities in the binding pattern of antisera to steroid hormones found on human and monkey spermatozoa. Assuming the intensity of fluorescence is proportional to the concentration of the hormone, concentrations of testosterone on the acrosomal and the postacrosomal regions were higher than levels of progesterone and estrogens. Spermatozoa with a "tapering head" had more hCG bound on the acrosomal and postacrosomal regions than spermatozoa with "normal head" (oval shaped). Correlating these findings to the functions of spermatozoa will require further studies.

Effect of clomiphene on the ponderal, histological and biochemical changes induced by stilbestrol and testosterone in the uterus of rats.

A comparative study on the effects of clomiphene on the stilbestrol- and testosterone-induced changes in the weight, histology and biochemical constituents of the uterus was undertaken. Clomiphene counteracted the stilbestrol-induced increase in the weight, and the absolute contents of the glycogen, protein and RNA of the uterus, possibly by competitive inhibition. It failed to prevent such changes induced by testosterone: on the contrary, there were some additive effects. The results of the present study indicate that the binding sites for the estrogen and the androgen in the uterus are different.

Effects of STS-557 and 20 Aet-1 on sperm functions and serum level of testosterone in bonnet monkey (Macaca radiata).

Treatment of adult male bonnet monkeys with STS-557 (17 alpha-cyanomethyl 17 beta-hydroxy estra-4, 9(10)-dien-3-one; 12 mg/monkey daily for 15 weeks; i.m.) reduced the sperm count from the 9th week, leading close to azoospermia on the 2nd post-treatment week which persisted until the 10th post-treatment week and normalcy was restored on the 13th post-treatment week. The sperm motility was reduced from the 9th week of treatment to 8th post-treatment week. The fertilizing ability of spermatozoa (formation of swollen sperm head or pronuclei in zona-free hamster eggs) was abolished from the 8th week of treatment to the 7th post-treatment week. The serum level of testosterone was reduced from the 2nd to 12th week of treatment (data on subsequent weeks were not collected). When 20 Aet-1 (testosterone-trans-4-n butyl cyclohexyl carboxylate; 40 mg/monkey, single; i.m.) was administered on the first day of STS-557 treatment, the sperm count was reduced from the 10th week, with near azoospermia ensuing on the 13th week which continued until the 8th post-treatment week; recovery was observed on the 12th post-treatment week. The motility was low from the 8th week of treatment to 6th week after withdrawal of treatment. The fertilizing ability of spermatozoa was abolished from the 8th week to the 12th week of treatment (data on last week of treatment were not collected). The serum level of testosterone was maintained within normal range except on the 2nd, 6th and 12th week of treatment. 20 Aet-1 alone had no significant effect on these parameters.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of STS 557 on semen and sperm functions in bonnet monkey (Macaca radiata)--a preliminary report.

Intramuscular administration of STS 557 (17 alpha C-cyanomethyl-17 beta-hydroxy-estra-4,9(10)-diene-3-one) at a daily dose of 12 mg/monkey for 8 weeks induced severe oligospermia to complete azoospermia by the 9th week of initiation of treatment with concomitant decrease in the serum level of testosterone. The sperm motility was reduced from the 3rd week of treatment. Complete recovery was observed by the 14th week after withdrawal of treatment. STS 557 may have the potentiality to be used as a chemical contraceptive in the male.

Contraceptive potentiality of STS-557--a feasibility study in male bonnet monkey (Macaca radiata).

STS-557 (17 alpha-cyanomethyl-17 beta-hydroxy-estra-4, 9(10)-diene-3-one) was administered (i.m.) to two groups (4 in each group) of adult male bonnet monkeys at a daily dose of 10 mg/monkey for 12 weeks (first group) and 5 mg/monkey for 14 weeks (second group). Treatment with the 10 mg dose resulted in a significant decline in the count, motility, acrosin and hyaluronidase activities and the fertilizing ability (zona-free hamster egg penetration assay) of spermatozoa by the 6th week of initiation of treatment. The circulating level of STS-557 was low after one week and increased from the 2nd week of treatment when the serum level of testosterone was significantly reduced. Complete recovery was observed by the 11th week after withdrawal of treatment. The treatment with the 5 mg dose had minor and inconsistent effect on the motility, hyaluronidase and acrosin activity, and the fertilizing ability of spermatozoa in addition to the blood level of testosterone. STS-557 may have the potentiality to be used as a chemical contraceptive in the male but compensation for the reduced level of blood testosterone may be necessary.

Use of oxide minerals to abate fluoride from water.

The removal of fluoride from aqueous solutions has been investigated using various oxide ores such as refractory grade bauxite, feed bauxite, manganese ore, and hydrated oxides of manganese ores (WAD). The refractory grade bauxite showed promising results. The studies were carried out as functions of contact time, pH, concentration of adsorbents, concentration of adsorbate, and temperature. The adsorption was rapid during the initial 5 min but equilibrium was attained within 120 min. The adsorption followed first-order kinetics. The present system followed the Langmuir adsorption isotherm model. Various thermodynamic parameters such as free energy, enthalpy, entropy, and equilibrium constants were calculated. The isosteric heat calculations showed that the adsorption process followed a heterogeneous model.

Effect of estradiol-17beta & estriol-3-methyl ether on spermatozoa & genital organs of rats.

PIP: The effect of estradiol-17beta (E2) and estriol-3-methyl ether on spermatozoa and genital organs was investigated in rats. The motility pattern of spermatozoa in epididymis and vas deferens was adversely affected by both treatments in intact rats. The number of spermatozoa in cauda epididymis was significantly (p less than .05) reduced after treatment with estriol-3-methyl ether, however, the combined treatment had a more severe effect. Sperm transport was accelerated after both treatments in castrated rats. No effect was observed on weight and gross histology of testis. Epididymis weight was reduced only after combined treatment. The weight of seminal vesicles, ventral prostate, and vas deferens was reduced after both treatments. However, in castrated rats there was a transient increase in the weight of vas deferens and seminal vesicles after 7 days of E2 treatment while ventral prostate weight was reduced.^ieng

Effect of fertility regulating agents on motility and zona-free hamster egg penetration by spermatozoa of bonnet monkey.

Administration of STS-557 (17 alpha-cyanomethyl-17 beta-hydroxyestra 4,9(10)-dien-3-one; 12 mg/monkey daily) for 4 weeks either alone or in combination with 20 Aet-1 (testosterone-trans-4-n-butyl cyclohexyl carboxylate; code CDB 1781; 40 mg/monkey single administration) had no significant effect on motility and zona free hamster egg penetration by spermatozoa of bonnet monkey, but continuation of the treatment for 12 weeks reduced (in one monkey treated with STS-557) or abolished (one treated with STS-557 and two with STS-557 + 20 Aet-1) the motility as well as zona-free hamster egg penetration (by spermatozoa of all treated monkeys). Motility and the ability to penetrate zona-free hamster egg returned to normalcy after 10 weeks of withdrawal of treatments. Active immunization of monkeys with ovine FSH (4 weeks after booster) had no adverse effect on motility of spermatozoa but none of the zona-free hamster eggs was fertilized. The correlation between motility and the capacity to penetrate the zona-free hamster eggs by monkey spermatozoa varies with the treatment. Such correlation was apparent in monkeys treated with STS-557 but not in monkeys immunized with ovine FSH.

Sperm production and fertility of bonnet monkeys (Macaca radiata) following immunization with ovine follicle stimulating hormone.

Adult male bonnet monkeys were rendered oligospermic but not azoospermic following active immunization with ovine follicle stimulating hormone. The percentage of sperms in the semen having good motility was reduced with a concomitant increase in the sperm ATPase activity. Eight out of 10 immunized monkeys failed to impregnate females of proven fertility after mating for consecutive three cycles while the remaining two impregnated the cohabitated females during the third cycle at a time when the antibody titer was reduced. Active immunization with ovine follicle stimulating hormone may not produce complete azoospermia but renders adult male monkeys infertile provided sufficient antibody titer is maintained.

Effects of intravasal nylon thread on the spermatozoa & reproductive organs in rats.

PIP: An investigation was undertaken in rats to study the effects of intr avasal thread (IVT) on the spermatozoa in the vas deferens and reproduct ive organs at various intervals after IVT insertion. The quantity of sperm was slightly reduced and motility was greatly reduced in the distal portion of the vas. The percentage of head and tail separation of sperm in the distal vas decreased with time. The quantity of sperm always remained the same in the cauda epididymis although the percentage of motile sperm decreased at 1 and 6 months, but not at 9 months, after IVT insertion. Following IVT insertion there was insignificant change in the weight of the testis, epididymis, ventral prostate, and seminal vesicles and alkaline phosphatase activity in the ventral prostate. Although cause and significance of these findings are unclear, the sialic level in the epididymis was significantly reduced in all groups bearing IVT. The presence of IVT apparently causes a change to occur in the epididymis, but it is unknown whether this affects sperm maturation.^ieng

Effect of synthetic progestins on the uptake of labelled testosterone and estradiol-17beta by the accessory genital organs of male rats.

PIP: The effect of pretreatment with norethindrone (NE) or 17-hydroxyprogesterone caproate (17-OHPC) on the uptake of tritiated testosterone and estradiol-17beta by the accessory sex organs of castrated and intact rats was investigated. A selective in vivo increase in the incorporation of tritiated testosterone and estradiol-17beta was observed at 48 hours after castration. The uptake of testosterone was greatest in the epididymis, while the maximum incorporation of estradiol-17beta was by the vas deferens. Pretreatment with NE or 17-OHPC decreased the incorporation of testosterone by all the accessory organs of castrated rats. NE decreased the incorporation of tritiated estradiol-17beta in the epididymis and seminal vesicles only, while 17-OHPC decreased the uptake in all accessory organs.^ieng

Differential response of testis and serum gonadotropins to testosterone in rats treated with a gonadotropin releasing hormone antagonist or estradiol 17 beta.

Adult rats treated with a GnRH antagonist (Ac D2Nal1, D4Cl Phe2, DTrp3, DArg6, DAla10 GnRH; code: 103-289-10, National Institutes of Health, USA) for 5 weeks (250 micrograms/kg b.w) showed multiple degrees of impairment and atrophy of the genital organs concomitant with decreased serum levels of testosterone, LH and FSH. Inhibition of spermatogenesis was characterized by germ cell degeneration and overall decline in different cell numbers and in particular, spermatids of any kind were completely absent. Testosterone supplementation (60 micrograms/rat/day, sc) to GnRH antagonist treated rats, for the same period, significantly elevated the weights of the sex organs, and the serum levels of hormones. Spermatogenesis was improved both qualitatively and quantitatively; albeit failed to be restored back to control levels. Treatment with estradiol 17 beta (1 microgram/rat/day) for 5 weeks had insignificant effect on spermatogenesis but the weights of the genital organs (seminal vesicles by 19% and ventral prostate by 40%) and the levels of serum hormones (LH by 24%, FSH 22% and testosterone by 25%) were otherwise reduced. Administration of testosterone either alone or in combination with estradiol 17 beta had only a marginal effect on spermatogenesis or on other reproductive parameters. The results indicate a positive shift in the response of the testis and serum levels of gonadotropins to testosterone supplementation in rats treated with either GnRH antagonist or estradiol 17 beta.

Effect of intra-uterine administration of dimethylsulfoxide on fertility of rats.

A single intra-uterine injection of 60 microliters of dimethylsulfoxide prevented implantation when administered before mating and induced resorption of the conceptus in rats when given during early pregnancy.

Cholesterol and ascorbic acid in semen under varied clinical conditions.

PIP: Fresh semen samples were collected from a total of 269 individuals attending the Family Welfare Planning Clinic. 29 had azoospermia following vasectomy, 12 had azoospermia due to spermatogenic defect, and the remainder had varying grades of sperm density and motility. Physical and morphological evaluations of the semen samples were made. Ascorbic acid was estimated in whole semen by the method of Roe an Kuether. Total cholesterol in plasma was determined. The total cholesterol concentration showed a gradual decline in the group with sperm count of 1-20 million/ml to the lowest level in the group with a count of 61-80 million. It then increased progressively up to the counts of 101-120 million/ml. Ascorbic acid concentration was significantly higher in patients with azoospermia due to defective spermatogenesis. The azoospermic specimens of vasectomized individuals contained significantly higher level of ascorbic acid than in dysspermatogenic azoospermia. The percentages of motile sperm in samples were unrelated to either cholesterol or ascorbic acid concentrations.^ieng

Effect of estradiol-17-beta on spermatozoa of castrated rats.

PIP: The effect of estradiol-17-beta on spermatozoa of castrated rats was investigated. Rats received daily im injections of .02 mcg estradiol-17-beta/100 gm body weight, 2 mcg testosterone/100 (gm body weight, or estradiol plus testosterone for 1, 2, 3, 4, or 5 weeks from the day after castration. Administration of estradiol maintained the normal motility pattern of spermatozoa of some parts of the epididymis and vas deferens for 7 days but by 21 days spermatozoa were absent in the whole tract. Testosterone restored the normal pattern of motility which was maintained for 14 days. The combined treatment also maintained the normal motility pattern up to 14 days. Estradiol accelerated and testosterone retarded sperm transport in the epididymis and vas deferens. Vas deferens and seminal vesicle weight increased after estradiol. Only epididymis weight increased after testosterone and only the seminal vesicle weight increased after combined treatment. These results indicate that estrogen may play a major role in the maintenance of the normal motility pattern of spermatozoa in the epididymis and vas deferens and in sperm transport in combination with androgen.^ieng