Impact of Italian legislation regulating assisted reproduction techniques on ICSI outcomes in severe male factor infertility: a multicentric survey.

BACKGROUND: In 2004, a law regulating assisted reproduction techniques (ART) was passed in Italy. The new rules allow for the formation and transfer of a maximum of three embryos at one time, whereas embryo selection and embryo storage are prohibited. The aim of this study is to evaluate the impact of these restrictions on ICSI outcome in couples affected by severe male factor infertility. METHODS: Thirteen Italian ART Units were involved in this study. Data were collected on ICSI cycles performed during 2 years before (control group) and 2 years after (study group) the enforcement of the law. Only cases of obstructive azoospermia (OA), non-obstructive azoospermia (NOA) and severe oligoastenoteratozoospermia (OAT) (sperm count

Sociability and preference for social novelty in five inbred strains: an approach to assess autistic-like behavior in mice.

Deficits in social interaction are important early markers for autism and related neurodevelopmental disorders with strong genetic components. Standardized behavioral assays that measure the preference of mice for initiating social interactions with novel conspecifics would be of great value for mutant mouse models of autism. We developed a new procedure to assess sociability and the preference for social novelty in mice. To quantitate sociability, each mouse was scored on measures of exploration in a central habituated area, a side chamber containing an unfamiliar conspecific (stranger 1) in a wire cage, or an empty side chamber. In a secondary test, preference for social novelty was quantitated by presenting the test mouse with a choice between the first, now-familiar, conspecific (stranger 1) in one side chamber, and a second unfamiliar mouse (stranger 2) in the other side chamber. Parameters scored included time spent in each chamber and number of entries into the chambers. Five inbred strains of mice were tested, C57BL/6J, DBA/2J, FVB/NJ, A/J and B6129PF2/J hybrids. Four strains showed significant levels of sociability (spend- ing more time in the chamber containing stranger 1 than in the empty chamber) and a preference for social novelty (spending more time in the chamber containing stranger 2 than in the chamber containing the now-familiar stranger 1). These social preferences were observed in both male and female mice, and in juveniles and adults. The exception was A/J, a strain that demonstrated a preference for the central chamber. Results are discussed in terms of potential applications of the new methods, and the proper controls for the interpretation of social behavior data, including assays for health, relevant sensory abilities and motor functions. This new standardized procedure to quantitate sociability and preference for social novelty in mice provides a method to assess tendencies for social avoidance in mouse models of autism.

Automated apparatus for quantitation of social approach behaviors in mice.

Mouse models of social dysfunction, designed to investigate the complex genetics of social behaviors, require an objective methodology for scoring social interactions relevant to human disease symptoms. Here we describe an automated, three chambered apparatus designed to monitor social interaction in the mouse. Time spent in each chamber and the number of entries are scored automatically by a system detecting photocell beam breaks. When tested with the automated equipment, juvenile male C57BL/6J mice spent more time in a chamber containing a stranger mouse than in an empty chamber (sociability), similar to results obtained by the observer scored method. In addition, automated scoring detected a preference to spend more time with an unfamiliar stranger than a more familiar conspecific (preference for social novelty), similar to results obtained by the observer scored method. Sniffing directed at the wire cage containing the stranger mouse correlated significantly with time spent in that chamber, indicating that duration in a chamber represents true social approach behavior. Number of entries between chambers did not correlate with duration of time spent in the chambers; entries instead proved a useful control measure of general activity. The most significant social approach behavior took place in the first five minutes of both the sociability and preference for social novelty tests. Application of these methods to C57BL/6J, DBA/2J and FVB/NJ adult males revealed that all three strains displayed tendencies for sociability and preference for social novelty. To evaluate the importance of the strain of the stranger mouse on sociability and preference for social novelty, C57BL/6J subject mice were tested either with A/J strangers or with C57BL/6J strangers. Sociability and preference for social novelty were similar with both stranger strains. The automated equipment provides an accurate and objective approach to measuring social tendencies in mice. Its use may allow higher-throughput scoring of mouse social behaviors in mouse models of social dysfunction.

Pulmonary oedema caused by "liquid ecstasy" ingestion.

In this case report we describe the first recorded case of alveolar gas exchange impairment caused by "liquid ecstasy" ingestion, and perhaps inhalation, by a 4 year old child. The pulmonary gas diffusion disturbance was sufficiently prolonged to raise the suspicion of a direct toxic effect on the alveolar-capillary membrane.

Synthesis, biological evaluation, and pharmacophore generation of new pyridazinone derivatives with affinity toward alpha(1)- and alpha(2)-adrenoceptors.

A series of new pyridazin-3(2H)-one derivatives (3 and 4) were evaluated for their in vitro affinity toward both alpha(1)- and alpha(2)-adrenoceptors by radioligand receptor binding assays. All target compounds showed good affinities for the alpha(1)-adrenoceptor, with K(i) values in the low nanomolar range. The polymethylene chain constituting the spacer between the furoylpiperazinyl pyridazinone and the arylpiperazine moiety was shown to influence the affinity and selectivity of these compounds. Particularly, a gradual increase in affinity was observed by lengthening the polymethylene chain up to a maximum of seven carbon atoms. In addition, compound 3k, characterized by a very interesting alpha(1)-AR affinity (1.9 nM), was also shown to be a highly selective alpha(1)-AR antagonist, the affinity ratio for alpha(2)- and alpha(1)-adrenoceptors being 274. To gain insight into the structural features required for alpha(1) antagonist activity, the pyridazinone derivatives were submitted to a pharmacophore generation procedure using the program Catalyst. The resulting pharmacophore model showed high correlation and predictive power. It also rationalized the relationships between structural properties and biological data of, and external to, the pyridazinone class.

Structure-activity relationships in a series of 8-substituted xanthines as A1-adenosine receptor antagonists.

A series of 8-substituted xanthines were synthesized and their affinity in vitro towards A1, A2A-adenosine receptors was evaluated by radioligand receptor binding assays. All compounds showed a greater affinity and selectivity towards the A1-adenosine receptor than theophylline. The compounds in which the n-proyl group is in 1-position of the xanthine nucleus and the pyridazinone system in 8-position is linked through a chain of two or four carbon atoms, showed the highest affinity and selectivity.

Synthesis and alpha(1)-antagonist activity of derivatives of 4-chloro-5-[4-[2-(2-methoxyphenoxy)-ethyl]-1-piperazinyl]-3(2H)-pyrid azinone.

The synthesis and evaluation of the biological activity of new 4-chloro-5-¿4-[2-(2-methoxyphenoxy)-ethyl]-1-piperazinyl¿-3(2H)-pyrid azinone derivatives are reported. The blocking activity of these compounds was determined on the pre- and postsynaptic alpha-adrenoceptors of isolated rat vas deferens.

Synthesis of new pyridazinone derivatives and their affinity towards alpha1-alpha2-adrenoceptors.

A series of 3(2H)-pyridazinone derivatives was evaluated for their affinity in vitro towards alpha1-alpha2-adrenoceptors by radioligand receptor binding assays. All target compounds showed good affinities for the alpha1-adrenoceptor (with Ki values in the subnanomolar range), and a gradual increase in affinity was observed by increasing the polymethylene chain length of this series up to a maximum of six and seven carbon atoms, when the fragment 4-[2-(2-methoxyphenoxy)-ethyl]-1-piperazinyl is linked in 5 position of the 3(2H)-pyridazinone ring, while a slight decrease was found for the higher homologues. Increasing the chain length when the 4-[2-(2-methoxyphenoxy)-ethyl]-1-piperazinyl group is linked in 6 position of the 3(2H)-pyridazinone ring, had a different effect: there is the highest affinity when the polymethylene chain is of four carbon atoms. The alkylic chain, a spacer between the two major constituents of the molecule, can influence the affinity and the selectivity.

Comparative molecular field analysis of some pyridazinone-containing alpha1-antagonists.

Diverse series of piperazines linked at N1 to 4, 5, or 6 positions of 3-(2H)-pyridazinone ring and at N4, by a suitable alkyl spacer, to some classical alpha1-adrenoceptor pharmacophore moieties, were tested in vitro for their alpha1-adrenoceptor antagonist activity. The modeling of their biological activity (pKb) by comparative molecular field analysis led to the development of a statistically significant partial least squares (PLS) model able to detect at 3-D level the main physicochemical interactions responsible for alpha1-adrenoceptor antagonist activity.

In vitro release of azidothymidine (AZT) by ceramic drug delivery systems.

Conventional methods of administration of 3-azido-2,3-dideoxythymidine (AZT) to treat AIDS patients is associated with negative side effects, including toxic effects on bone marrow. This study was designed to investigate the capability of tricalcium phosphate (TCP) and alumino-calcium-phosphorous oxide (ALCAP) drug delivery systems to release AZT in a sustained manner in an in vitro environment. A total of 35 TCP and 35 ALCAP ceramic capsules were used in this experiment. The final densities of the ceramics were 1.68 +/- 0.11 and 1.58 +/- 0.08 g/cm3 for ALCAP and TCP, respectively. Tricalcium phosphate ceramics were distributed into four groups. Each ceramic in groups I, II and III (n = 10) was loaded with 20, 40, or 60 mg of AZT in powder form, respectively. Group IV ceramics (n = 5) were left empty and served as a control group for possible corrections of absorbance readings. A similar protocol was followed for ALCAP ceramic capsules unless otherwise indicated. All TCP and ALCAP ceramic capsules were sealed at both ends with Silastic Medical Adhesive Silicon Type A (Dow Corning, Midland, MI). Each empty capsule and those loaded with AZT were sealed and suspended in a capped and sealed individual serum bottles containing 100 mls of phosphate buffered saline (pH 7.4), and agitated at 100 cycles per min in a water bath at 37 degrees C. The amount of AZT released from the ceramic reservoir into the buffered medium was measured spectrophotometrically (265 nm). Release of AZT from the ceramic capsule was monitored until the contents of the capsules were depleted.(ABSTRACT TRUNCATED AT 250 WORDS)

Sustained delivery of 3H-thymidine by means of ceramic capsules in rats.

The use of both oral and intravenous administration of 3-azido-2,3-dideoxythymidine (AZT) to treat AIDS patients is associated with negative side effects, including toxic effects on bone marrow. This study was designed to investigate the release of deoxynucleoside thymidine, the normal counterpart of AZT, by means of ALCAP ceramic implantable capsules in rats. ALCAP capsules (1.79 +/- 0.03 g/cm3) were impregnated with a solution of polylactic acid (2% wt/vol) in chloroform. Each capsule was loaded with 40 mg of unlabeled thymidine and 10 microCi of 3H-thymidine. A total of 60 SD-male rats were distributed into three groups (sham, control, and experimental). Each experimental rat was implanted intraperitoneally with a thymidine loaded ceramic capsule. Blood samples were collected at weekly intervals from each rat, for eight weeks, via the tail artery. A total of 351 +/- 9.12, 308 +/- 11.29, 350 +/- 21.28 and 322 +/- 31.76 micrograms/ml of thymidine was released from the ceramic capsule at the end of 14, 28, 42, 56, days, respectively. The data obtained shows that ALCAP ceramic capsules can deliver thymidine in a controlled, safe, and sustained manner in rats for a minimum duration of 120 days. It also suggests that ALCAP capsules probably can be used to deliver nucleosides, such as AZT, in a sustained manner for long durations in mammalian systems, including humans.

Effects of delivering different amounts of dihydrotestosterone by ceramic implants on the reproductive system of male rats.

Administration of drugs or hormones to patients, orally or by injections, results in a pulsatile delivery of the drug. The purpose of this study was to determine whether dihydrotestosterone (DHT), delivered in a sustained manner by ALCAP ceramic capsules, can regulate fertility in male rats. Sixty rats were distributed equally into 4 groups. Each of the 15 capsules (1.60 +/- 0.01 gm/cm3) in groups I, II and III was loaded with 30, 60, and 90 mg DHT, respectively and implanted in rats. Rats in group IV served as controls and were implanted with empty capsules. Five rats from each group were euthanized at the end of 1, 2, and 3 months. The data obtained showed that the weights of vital organs of rats in all groups did not differ significantly. At the end of 3 months, accessory reproductive organs (ARO) in group I and group II rats showed significantly lower sperm counts, but maintained the weights of ARO. Group III rats did not show a significant decrease in the sperm counts or weights of ARO in comparison to values obtained for control rats. The data obtained suggest that DHT, delivered in appropriate amounts by ceramic capsules, can be used to regulate fertility in mammals.