Activity of delta8- and delta9-tetrahydrocannabinol and related compounds in the mouse.

The 11-hydroxy metabolites of Delta(8).- and Delta(9)-tetrahydrocannabinol are more active than the parent compounds when administered to mice by either the intravenous or intracerebral route. Both Delta(8)- and Delta(9)-tetrahydrocannabinol are rapidly and extensively metabolized by the liver and not by the brain. The hypothesis that the 11-hydroxy metabolites may be the active form of tetrahydrocannabinol is discussed

Psychosocial outcomes of bone marrow transplant for individuals affected by Mucopolysaccharidosis I Hurler Disease: patient social competency.

AIM: To explore the frequency with which children and young people participate in social activities with peers, when they are affected by Mucopolysaccharidosis I Hurler Disease (MPS IH) post bone marrow transplant (BMT). This was investigated in relation to patient age, and in comparison with a normative sample. Patient withdrawal, adaptive and social skills are also described in terms of patient age and in comparison with a normative sample. METHOD: Forty-four individuals affected by MPS IH post BMT participated in this investigation. Their ages ranged from 16 months to 25 years. Semi-structured interviews with patients' mothers were utilized, which included the Behaviour Assessment System for Children and a socialization questionnaire. Normative data for the socialization questionnaire were collected from 46 mothers of children not affected by chronic illness or disability. RESULTS: A one-way ANOVA revealed that children not affected by disability or chronic illness (mean = 20.63) participated overall in social activities more frequently than children aged under 12 years (mean = 14.87) and over 12 years (mean = 13.25) who were affected by MPS IH post BMT (F = 21.01, P < 0.001). Young people aged 12 years and over affected by this condition were found to participate in social activities the least. A pattern also emerged, which indicated greater withdrawal and less well-developed adaptive and social skills for MPS IH patients aged 2.5-5 years and 12 years and over, but scores were within the normal range for those aged 6-11 years. CONCLUSION: These data illustrate a lack of social competency and a tendency towards inhibition and withdrawal in this patient group, particularly among the very young children and the adolescents and young adults. Further research is needed to explore these issues longitudinally and to examine the role played by the family, and indeed disability, in the quality and quantity of social interaction experienced by these individuals.

Synthesis of 11 beta,13 beta- and 13 beta,16 beta-propano steroids: probes of hormonal activity.

Syntheses of 11 beta,13 beta- and 13 beta,16 beta-propano derivatives of 17 alpha-ethynyl-17 beta-hydroxygon-4-en-3-one are described. The 13 beta,16 beta bridge was constructed by intramolecular alkylation of the C-16 enolate anion from 3-methoxy-13 beta-[3'-(tosyloxy)propyl]gona-3,5-dien-17-one, the latter being obtained via Birch reduction of both aryl groups of 17 beta-hydroxy-3-methoxy-13 beta-(3'-phenoxypropyl)gona-1,3,5(10),8-tetraene (1). The 11 beta,13 beta bridge was constructed by Prins cyclization of 17 beta-acetoxy-3-methoxy-13 beta-(3'-oxopropyl)gona-1,3,5(10),9(11)-tetraene, itself obtained via Birch reduction of only the side-chain aryl group of 1. Binding affinities of certain of these compounds and substituted 13 beta-propyl derivatives of 17 alpha-ethynyl-17 beta-hydroxygon-4-en-3-one for the uterine cytosol receptor of progesterone are reported, and the origin of the high progestational activity of norgestrel and 11 beta-substituted progestins is discussed.

Synthesis, antifertility activity, and protein binding affinity of 7(8 leads to 11 alpha)abeo steroids.

A series of 7(8 leads to 11 alpha)abeo steroids was synthesized by a modification of the previously described total synthesis of this class of compounds and evaluated for biological activity. In general, there was a marked reduction in the relative binding affinities of these compounds for the rabbit uterus estrogen and progestin receptor proteins. None of the compounds which were subjected to uterotropic, antiuterotropic, postcoital, progestational, antiprogestational, or antiandrogenic assays showed any significant activity.

PIP: Synthesis, antifertility activity, and protein binding afinity of 7(8 to 11alph) abeo-estranes and -pregnanes are described. There was a marked reduction in the relative binding affinities of these compounds for the rabbit uterus estrogen and progestin receptor proteins. None of the compounds subjected to uterotropic, antiuterotropic, postcoital, progestational, antiprogestational, or antiandrogenic assays revealed any marked activity.

Esters and lactones of phenolic amino carboxylic acids: prodrugs for iron chelation.

The new iron chelator N,N'-bis(2-hydroxyphenyl)ethylenediamine-N,N'-diacetic acid (1), its dilactone 2, N,N'-bis(2-hydroxybenzyl)-2-hydroxypropylene-1,3-diamine-N,N'- diacetic acid (3), and its methyl ester lactone 4 and a series of esters of N,N'-bis(2-hydroxybenzyl)ethylenediamine-N,N'-diacetic acid (5) were prepared and their iron chelating efficacy and toxicity determined by using the hypertransfused mouse model of iron overload. The biological activities were compared with results obtained with use of the hypertransfused rat. Esterification enhanced the oral iron chelating activity but also increased toxicity. The diisopropyl ester of 5 exhibited the highest therapeutic index. In vitro measurements showed that the rate of ester hydrolysis at pH 7.5 increased by a factor of 10(4) in the presence of 5 X 10(-4) M ferric ion, which may account for the utility of esters and lactones as prodrugs. Seventeen other chelating agents were screened but showed no intraperitoneal or oral activity.

The biodegradability of polyester blends.

Blends of poly(epsilon-caprolactone) (PCL) and poly(L-lactic acid) (PLLA) with polyglycolic acid-co-L-lactic acid (PGLA) were prepared by three methods: compression moulding, coprecipitation, and solvent evaporation of a methylene chloride-in-water emulsion of the polymers. The rates of hydrolytic chain scission of each component of the blends were determined by deconvolution of GPC traces of samples maintained in phosphate buffer, pH 7.4, 37 degrees C, for up to 3000 h. The observed rates were dependent on the method of blending. For compression moulded blends, the rate of chain scission of PGLA was decreased and that of PCL and PLLA increased. A corresponding delay in the onset of weight loss was also observed. There was no evidence of blend miscibility.

Spin labels as a probe of the molecular environment of covalently bound ligands in an hydrophobic and an hydrophilic polymer.

The molecular motions of spin-labelled ligands covalently bound by spacer groups to an hydrophobic and an hydrophilic polymer matrix were evaluated by ESR spectroscopy. The ligands were prepared by alkylation of 4-N-methylamino-TEMPO with the omega-bromocarboxylic esters, Br(CH2)nCOOEt, n = 1,4 and 10. The hydrolysed esters were coupled to a cross-linked aminoethylated polyacrylamide hydrogel (n = 1,4,10) and to a surface hydroxylated elastomeric polyester (n = 4). The rotational correlation times (tau c) of the nitroxide label in the hydrogels were measured in the dry state and after exposure to water at pH 4, 7.4 and 10.5. The tau c of the nitroxide label was insensitive to the length of the spacer group and to the degree of protonation of the tertiary amino group of the ligand. There was no evidence of self-association of the ligand and spacer, or more than a single phase within the polyacrylamide hydrogel. The tau c of the nitroxide labelled polyester was similarly insensitive to pH, but was sensitive to organic solvents. The low mobility of the spin label and its high concentration were consistent with the spin label being covalently bound within the hydrophobic polyester matrix to a depth of at least 5 microns.

Aliphatic polyesters II. The degradation of poly (DL-lactide), poly (epsilon-caprolactone), and their copolymers in vivo.

The mechanisms of biodegradation of poly (DL-lactide), poly (epsilon-caprolactone), and copolymers of epsilon-caprolactone with DL-dilactide, delta-valerolactone, and DL-epsilon-decalactone in rabbit were shown to be qualitatively similar. However, the rate of the first stage of the degradation process, non-enzymatic random hydrolytic chain scission, varied by an order of magnitude and was dependent on morphological as well as chemical effects. Weight loss was generally not observed until the molecular weight had decreased to 15,000 or less. Poly (DL-lactide) differed from the other polyesters studied, the rate of chain scission increasing after the commencement of weight loss. The rate of weight loss was greater and the period prior to weight loss was shorter when the comonomer content of copolymers of epsilon-caprolactone was sufficient to reduce the melting point of epsilon-caproate sequences to body temperature.

Adjunctive intravaginal metronidazole for the prevention of postcesarean endometritis: a randomized controlled trial.

OBJECTIVE: To estimate the efficacy of preoperative administration of intravaginal metronidazole for the prevention of postcesarean endometritis. METHODS: This double-masked, placebo-controlled randomized trial included patients of at least 24 weeks' gestation undergoing cesarean deliveries for various indications. Patients were randomized to receive either 5 g of metronidazole gel intravaginally or matching placebo before the initiation of the cesarean. All patients underwent surgical cleansing of the abdomen, and most received prophylactic antibiotics after cord clamping. Patients with chorioamnionitis and/or suspected allergy to metronidazole were excluded. For a two-sided alpha of 0.05 and beta of 0.20 (80% power), 120 subjects were required in each group. The main outcome variable was the incidence of postcesarean endometritis. Secondary outcome variables included presence of febrile morbidity, wound infection, days on antibiotics, and length of postpartum hospitalization. Neonatal outcomes included birth weight, Apgar scores less than 7 at 5 minutes, umbilical arterial pH less than 7.16, admission and length of stay in the neonatal intensive care unit, and length of hospital stay. RESULTS: Of 112 patients receiving metronidazole, eight (7%) developed postcesarean endometritis, compared with 19 of 112 (17%) of those receiving placebo gel (relative risk 0.42, 95% confidence interval 0.19, 0.92). No significant differences were noted between treatment groups with respect to the other outcome variables. CONCLUSION: The preoperative administration of 5 g of intravaginal metronidazole gel appears to reduce the incidence of postcesarean endometritis.

Prophylactic amnioinfusion for intrapartum oligohydramnios: a meta-analysis of randomized controlled trials.

OBJECTIVE: To evaluate the effectiveness of intrapartum prophylactic amnioinfusion in pregnancies complicated by oligohydramnios. DATA SOURCES: Randomized controlled trials of prophylactic amnioinfusion in women with oligohydramnios were identified using computerized databases, index reviews, and references cited in original studies and review articles. METHODS OF STUDY SELECTION: We evaluated, abstracted data from, and analyzed randomized studies of prophylactic intrapartum amnioinfusion in women with oligohydramnios. In every study the group allocation was based exclusively on presence of oligohydramnios. Only published studies with clearly documented outcome data were included. The quality of each trial was evaluated for methodology, inclusion and exclusion criteria, adequacy of randomization, amnioinfusion protocols, definition of outcomes, and statistical analyses. The trials were evaluated concerning cesarean deliveries for fetal heart rate (FHR) abnormalities, overall cesarean rates, acidemia at birth, intrapartum fetal heart rate abnormalities, Apgar scores under 7 at 5 minutes, and postpartum endometritis. TABULATION, INTEGRATION, AND RESULTS: Thirty-five studies were identified, of which 14 met the inclusion criteria for this systematic review. They included 1533 patients, 793 in the amnioinfusion group, and 740 controls. Odds ratios (OR) with their 95% confidence intervals (CI) for each outcome were calculated. We calculated an estimate of the OR and risk difference for dichotomous outcomes using random and fixed-effects models. A test of homogeneity was done across studies. Women with oligohydramnios who received intrapartum amnioinfusion had lower incidence of cesarean for FHR abnormalities (OR 0.23; 95% CI 0.15, 0.35). Intrapartum amnioinfusion also was associated with lower overall rates of cesarean deliveries (OR 0.52; 95% CI 0.40, 0. 68), acidemia at birth (OR 0.40; 95% CI 0.30, 0.55), FHR abnormalities during labor (OR 0.24; 95% CI 0.17, 0.34), and Apgar scores under 7 at 5 minutes (OR 0.52; 95% CI 0.29, 0.91). Postpartum endometritis rates were similar among the study groups. CONCLUSION: In the presence of oligohydramnios, prophylactic intrapartum amnioinfusion significantly improves neonatal outcome and lessens the rate of cesarean delivery, without increasing the rate of postpartum endometritis.

Outpatient in vitro fertilization using transvaginal ultrasound-guided oocyte retrieval.

Of 80 patients who initiated in vitro fertilization cycles in an integrated outpatient facility, 61 had oocyte retrievals performed under local anesthesia with ultrasound-guided transvaginal aspiration of ovarian follicles. Ten patients conceived, all in the group of 49 who had human chorionic gonadotropin (hCG)-timed retrievals. Our results indicate that in vitro fertilization can be performed to a high standard with this method, with excellent patient acceptance and reduced cost without general anesthesia, laparoscopy, or in-hospital setting.

Effects of cryopreservation on bull sperm head morphometry.

Artificial insemination using cryopreserved semen is a common management tool of the contemporary livestock producer. However, cryopreservation is detrimental to sperm function and fertility, killing some 50% of the spermatozoa during the process. Prediction of cryopreservation damage from prefreeze samples remains elusive. Computer-automated sperm head morphometry was used in this study to determine the effects of cryopreservation on bovine sperm head morphometry. Semen was collected from 18 bulls and was divided. One portion was extended to 200 x 10(6) sperm/ml and a microscope slide was prepared, while the remaining portion was cryopreserved in a Triscitrate-yolk extender. After thawing, the cryopreserved samples were prepared on microscope slides. All slides were air dried and were stained with hematoxylin and rose bengal. The morphometric dimensions for length, width, width/length, area, and perimeter for a minimum of 200 sperm heads were analyzed from each slide by computer-aided sperm head morphometry analysis, and the mean measurements were recorded. Bull sperm heads were significantly (P < 0.01) smaller in cryopreserved spermatozoa than in the companion extended samples for length (8.56+/-0.07 vs. 8.63+/-0.08 microm), width (4.39+/-0.05 vs. 4.48+/-0.05 microm), area (28.42+/-0.07 vs. 29.14+/-0.08 microm), and perimeter (23.33+/-0.21 vs. 23.70+/-0.23 microm) for all bulls. Width/length was also different (0.513 vs. 0.519). In addition, differences (P < 0.05) were found within 14 of 18 bulls for at least four of the morphometric parameters. The percent change in measures after cryopreservation were correlated (P < or = 0.05) to the variability of the extended sample. Variations in sperm head measurements were lower (P < or = 0.05) in extended samples of the four bulls in which no changes occurred than in extended samples of the remaining 14 bulls. These data suggest that the variability in sperm head measurements of individual bulls, or ejaculates, may be an indicator of sperm cryosurvivability.

Sustained drug delivery systems II: Factors affecting release rates from poly(epsilon-caprolactone) and related biodegradable polyesters.

The release rates of several steroids from films and capsules of homopolymers and copolymer of epsilon-caprolactone, DL-lactic acid, and glycolic acid were measured in vitro and in vivo for up to 200 days. Relatively constant release rates from capsules (reservoir devices) were observed only under certain conditions. Factors that influence the drug release kinetics were evaluated. Release from poly(epsilon-caprolactone) and poly(epsilon-caprolactone-co-DL-lactic acid) was diffusion controlled. Release from poly(DL-lactic acid-co-glycolic acid) was associated with polymer degradation. Release from poly(DL-lactic acid) was very slow when diffusion controlled.

Changes in susceptibility of bovine sperm to in situ DNA denaturation during prolonged incubation at ambient temperature under conditions of exposure to reactive oxygen species and nuclease inhibitor.

Sperm were incubated for up to 9 days in the presence or absence of exogenous hydrogen peroxide, phenylalanine, catalase and aurintricarboxylic acid to assess the influence of reactive oxygen species and inhibition of deoxyribonucleases on sperm chromatin stability. The assessment of sperm DNA susceptibility to in situ acid denaturation by the sperm chromatin structure assay did not detect any difference in chromatin stability between sperm incubated for 9 days under aerobic and anaerobic conditions in a diluent called 14G. Exposure to exogenous hydrogen peroxide under both aerobic and anaerobic conditions and to phenylalanine under aerobic conditions (which produces hydrogen peroxide by a reaction catalysed by the aromatic amino acid oxidase present in sperm) was detrimental to sperm chromatin stability, increasing its DNA susceptibility to in situ acid denaturation over the incubation time. This effect was eliminated if catalase was present in the diluent. Inclusion of the general deoxyribonuclease inhibitor aurintricarboxylic acid in the diluent severely decreased sperm chromatin stability under both aerobic and anaerobic conditions. Aurintricarboxylic acid was mildly cytotoxic, as revealed by viability assessment, under aerobic, but not under anaerobic, incubation conditions. Exogenous hydrogen peroxide, either directly added to the diluent or generated through the enzymatic oxidation of phenylalanine, was detrimental to sperm motility and the integrity of the plasma membrane.

Intranasal absorption of granulocyte-colony stimulating factor (G-CSF) from powder formulations, in sheep.

Granulocyte-colony stimulating factor (G-CSF) was administered to sheep in three different nasal formulations and as a subcutaneous injection. The nasal formulations were: a solution containing L-alpha-lysophosphatidylglycerol (LPG), a powder formulation comprising small starch microspheres (SSMS) and a powder formulation comprising SSMS and LPG. Absorption of G-CSF was assessed directly by quantitation in plasma and indirectly by measurement of the pharmacodynamic response in terms of leucocyte and neutrophil counts. After the nasal delivery of the G-CSF powder formulation containing SSMS and LPG the absorption of G-CSF was significantly higher (P<0.01) than that from the simple nasal solution or the powder without the enhancer, but the resulting pharmacological response was not significantly different. The bioavailability of G-CSF from the powder formulation containing SSMS and LPG relative to the subcutaneous injection was 8.4% (+/-3.4). We also found that at the respective G-CSF doses investigated, the pharmacodynamic response of this nasal formulation, was similar to that obtained after the subcutaneous administration. The study indicates that the powder formulation containing enhancers could offer an alternative delivery route for G-CSF in the form of intranasal administration.

Computer automated morphometric analysis of bull sperm heads.

Morphological type classification of spermatozoa is an important component of the modern semen evaluation; however, current methods of analysis are subjective and highly variable between technicians. To reduce the subjectivity and thus variability of sperm morphology assessment, computer automated sperm head morphology analysis (ASMA) has been developed. Previous studies have shown the importance of standardizing ASMA procedures to optimize accuracy. The objective of this study was to standardize ASMA procedures for evaluating bull sperm heads. Semen from 10 fertile bulls was used to standardize procedures for optimal analysis of bull spermatozoa. Sample preparation methods, sperm staining methods and microscopic magnifications were compared. Semen samples that were diluted to a standard concentration of 200 x 10(6) sperm/ml were more efficiently analyzed than raw samples. A modified GZIN staining procedure, incorporating rose bengal as an acrosomal stain, was used for accurate ASMA at a magnification of x 60. The mean morphometric measurements for all bulls were the area (27.30 microM), perimeter (25.36 microM), length(8.65 microM), width(4.40 microM) and width/length (0.50). Within the analyses, coefficients of variation ranged from 3.45% for length to 8.52% for area. The ASMA system correctly digitized sperm heads 97% of the time. Results of this study indicate that bull sperm heads can be accurately analyzed using current standard procedures of ASMA technology.

Selected times of insemination with microencapsulated bovine spermatozoa affect pregnancy rates of synchronized heifers.

This experiment was designed to test whether spermatozoa encapsulated in an alginate poly-L-lysine matrix had an extended fertile life in vivo after insemination. Estrus was synchronized in 417 primiparous Friesian and Jersey heifers with a system based on a CIDR-B intravaginal device before the heifers were inseminated either during proestrus (24 h after device removal) or at estrus (48 h after device removal). Pregnancy rates to first inseminations did not differ between the 24 and 48 h inseminations (61 vs 60.6%) with liquid semen diluted in Caprogen (control) but differed with encapsulated semen (45.1 vs 68.6%). The difference in pregnancy rates between the 2 types of semen was more pronounced (P < 0.08) in the animals that were visually detected in estrus. The mean survival time of spermatozoa in the female reproductive tract following insemination at the 24-h insemination time was estimated to be 50 +/- 7.5 h. The increased pregnancy rate with insemination of encapsulated spermatozoa at 48 h could have been due to this process predisposing spermatozoa to capacitate soon after insemination.