The implementation of a hepatitis C testing service in community pharmacies: I-COPTIC consensus statement.

OBJECTIVES: This aimed to develop a blueprint for an effective community pharmacy Hepatitis C virus (HCV) testing service by producing a consensus statement. STUDY DESIGN: This was a modified Delphi process. METHODS: We recruited a heterogenous panel of experts (who had been involved in the setup or delivery of a community pharmacy HCV testing service) by purposive and chain referral methods. We had three rounds of a modified Delphi process. The first was a series of questions with free text responses and was analysed using thematic analysis, and the second and third were statements for the respondents to rate using a 7-point Likert scale. Consensus was predefined in a published protocol, and the results were reviewed by a public and patient involvement panel before the statement was finalised. RESULTS: We had 24 participants, including community and hospital-based pharmacists, local pharmaceutical committee members, charity representatives (Hepatitis C Trust), local clinical service lead, nurse specialists and doctors. The response rate of the first, second and third rounds were 100%, 96% and 88%, respectively. After the third round, we had 60 statements that reached consensus. We discussed the accepted statements with a patient and public involvement group. We used these statements to produce the I-COPTIC statement and a graphical summary. CONCLUSIONS: We developed a blueprint for the design of a gold standard community pharmacy HCV testing service. We believe this will support the successful implementation of community pharmacy testing for HCV. Community pharmacy testing is an important service to help achieve and maintain HCV elimination.

Preimplantation genetic screening outcomes are associated with culture conditions.

BACKGROUND: Although preimplantation genetic screening (PGS) is widely offered, there are contradictory reports on the clinical merit of this procedure. Any gain from embryo selection following aneuploidy screening must significantly outweigh the impact of the procedure. Variability of technical expertise in embryo biopsy, blastomere fixation, fluorescence in situ hybridization analysis, along with suboptimal laboratory quality control and inappropriate patient selection may impact PGS outcomes. METHODS: To investigate such effects, a total of 1508 stimulated in vitro fertilization (IVF) cycles were retrospectively analysed. During 2004, a significant change was made to the embryo culture media used. Clinical outcomes from cycles with PGS were compared prior to and after the change in media and compared with matched controls not utilizing PGS during the same period. RESULTS: Clinical PGS success rates were found to improve following the media change. For patients aged less than 40, clinical outcomes following PGS were significantly lower than those without PGS prior to the change, but became equivalent after the change. For patients >or=40 years and

Substituted 6,7-dihydroimidazo[1,2-a]purin-9(4H)-ones.

The synthesis of a series of substituted 6,7-dihydroimidazo[1,2-a]purin-9(4H)-ones is described. Several members of the series exhibit enhanced antiallergic and bronchodilator activity and reduced side effects as compared to theophylline. Structure-activity relationships and metabolic considerations are discussed for the series. Analogues substituted with a 4-(4-chlorobenzyl) moiety, such as 33 and 40, shown an optimal balance of antiallergic and bronchodilator activity and are of particular interest. Compound 33 is significantly more potent than theophylline against both metacholine- and antigen-induced bronchospasms, does not affect spontaneous motor activity, and shows minimal cardiovascular effects in the rat.

Synthesis and biological evaluation of 1-(1,2-benzisothiazol-3-yl)- and (1,2-benzisoxazol-3-yl)piperazine derivatives as potential antipsychotic agents.

Members of the series of title compounds were tested for potential antipsychotic activity in relevant receptor binding assays and behavioral screens. Structure-activity relationships within the series are discussed. Compound 24 (BMY 13859-1), a (1,2-benzisothiazol-3-yl)piperazine derivative, was selected for further study because of its potent and selective profile in primary CNS tests. It was active in the Sidman avoidance paradigm and blocked amphetamine-induced stereotyped behavior in dogs for up to 7 h. The compound's lack of typical neuroleptic-like effects in the rat catalepsy test and its failure to produce dopamine receptor supersensitivity following chronic administration indicate that it should not cause the movement disorders commonly associated with antipsychotic therapy. Although 24 has potent affinity for dopaminergic binding sites, its even greater affinity for serotonin receptors suggests that a serotonergic component may be relevant to its atypical profile. Compound 24 is currently undergoing clinical evaluation in schizophrenic patients.

Synthesis, biological profile, and quantitative structure-activity relationship of a series of novel 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors.

A series of 9,9-bis(4-fluorophenyl)-3,5-dihydroxy-8-(alkyltetrazol-5-yl)- 6,8-nonadienoic acid derivatives 1 were synthesized and found to inhibit competitively the enzyme 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase. The analogues having 1N-methyltetrazol-5-yl attached to the C8-position (3a, 4a, R1 = R2 = F) are the most active in suppressing cholesterol biosynthesis in both in vitro and in vivo models: the IC50 for the chiral form of 3a is 19 nM, Ki = 4.3 x 10(-9)M when Km for HMG-CoA is 28 x 10(-6) M;1 the ED50 (oral) value corresponding to the lactone derivative (4a, BMY 22089) is approximately 0.1 mg/kg. Further, BMY 21950 is nearly 2 orders of magnitude more active in parenchymal heptaocytes, from which most of the serum cholesterol originates, than in other cell preparations (such as spleen, testes, ileum, adrenal, and ocular lens epithelial cells; Table III). This apparent tissue specificity may be highly beneficial since the blocking of cholesterol biosynthesis in other vital organs could eventually lead to undesirable side effects. In addition to the chemical synthesis and biological evaluation, a theoretical study aimed at relating the HMG-CoA reductase inhibitory potency to the three-dimensional structure of the inhibitors was undertaken. With a combination of molecular mapping and 3D-QSAR techniques, it was possible to determine a logical candidate for the conformation of the bound inhibitor and to quantitatively relate inhibitory potency to the shape and size of both the binding site and the C8-substituent.

A prisoner with chronic myeloid leukemia.

The health care received by prisoners in the area of tissue and organ transplantation is not well discussed or documented. We encountered a prisoner with newly diagnosed chronic myelogenous leukemia who was a candidate for allogeneic bone marrow transplantation and had two HLA-identical siblings who were willing to donate bone marrow. Based on humanitarian, constitutional, and ethical considerations, we suggest that the prisoner (patient) should receive the same health care as individuals who are not incarcerated and that the costs of care should play no greater role for prisoners than for other members of society.

Manipulation of sperm before intracytoplasmic sperm injection improves fertilization rates.

OBJECTIVE: To determine the effect of sperm manipulation before intracytoplasmic sperm injection (ICSI) on fertilization rates. DESIGN: Three methods of sperm manipulation before direct ICSI were compared in two sibling oocyte trials. SETTING: In vitro fertilization unit within a teaching hospital. PATIENTS: Patients undergoing infertility treatment using ICSI. INTERVENTIONS: Oocytes were inseminated by ICSI. MAIN OUTCOME MEASURE: Fertilization rate. RESULTS: In the first trial, a standard manipulation technique gave a 67% fertilization rate compared with 64% with a minimal manipulation technique. In the second trial the standard technique gave a significantly greater fertilization rate compared with no prior manipulation (67% versus 45%). CONCLUSIONS: Manipulation of sperm before direct ICSI is not mandatory for fertilization but significantly improves the fertilization rate.

Fertilization rates using intracytoplasmic sperm injection are greater than subzonal insemination but are dependent on prior treatment of sperm.

OBJECTIVE: To determine a suitable method of sperm preparation for use in micromanipulation. To compare the fertilization rates of sibling oocytes inseminated by intracytoplasmic sperm injection (ICSI) and subzonal insemination (SUZI). DESIGN: Two methods of gamete micromanipulation to overcome male factor infertility were compared. Within this study, three trials were conducted to determine the most suitable method of sperm preparation. This method then was used to complete the study. SETTING: Procedures were performed in a teaching hospital research environment. PATIENTS: Ninety-six patients were recruited for this study, undergoing 99 stimulation cycles. INTERVENTIONS: Sibling oocytes were inseminated by subzonal sperm or intracytoplasmic injection. MAIN OUTCOME MEASURES: Fertilization, zygote development, and pregnancy rates. RESULTS: In the absence of manipulative pretreatment of sperm (trial 1), there was no difference in normal fertilization rates between ICSI and SUZI (19% and 25%, respectively). In the second trial there again was no sperm pretreatment for SUZI but, for ICSI, a polyvinylpyrrolidone (PVP) solution was used to reduce sperm velocity and the sperm tails were incised before injection. The fertilization rates were significantly different between ICSI (44%) and SUZI (17%). In the third trial, PVP was added to the sperm used for both types of insemination and the sperm tails also were incised for the ICSI insemination. Fertilization was again significantly different (16% for SUZI and 44% for ICSI). The trial 2 method of sperm preparation then was used to complete the study (trial 4) and confirmed the results of trial 2 (21% for SUZI and 42% for ICSI). The results suggest that ICSI can give improved fertilization compared with SUZI provided the sperm are treated before injection. No significant differences were found in the development rates of zygotes suitable for transfer or cryopreservation between the two micromanipulation methods.

Short-term corticosteroid treatment does not improve implantation for embryos derived from subzonal insertion of sperm.

A total of 111 patients undergoing treatment for male factor infertility using SUZI participated in the trial for one treatment cycle only. They were allocated to have either corticosteroid treatment to induce immunosuppression or placebo. An elevated beta-hCG was found in 25% (9 of 36) of patients having an ET and receiving treatment. The corresponding figure for the control group was 33% (10 of 30). On ultrasound examination, fetal hearts were found in 22% (8 of 36) and 20% (6 of 30), respectively, of patients having an ET. chi 2 analysis showed no difference in the results. When the results were analyzed on a per oocyte basis there was no significant difference in the implantation rate.

Subzonal insertion of multiple sperm is a treatment for male factor infertility.

OBJECTIVE: To determine whether subzonal insemination of multiple sperm is an effective treatment for male factor infertility. DESIGN: Subzonal insemination by micromanipulation of sperm and oocytes for male factor patients. PATIENTS: Two hundred eight patients were selected on previous fertilization history and/or semen parameters. They could be assigned to one of three groups: group A had previous IVF failure, group B had a low fertilization rate with previous IVF, and group C had insufficient sperm for IVF. INTERVENTIONS: Oocytes were inseminated by subzonal insertion of multiple sperm. MAIN OUTCOME MEASURES: Fertilization and pregnancy rates. RESULTS: There have been 208 patients treated through 332 treatment cycles. Of 2,789 oocytes inseminated, 784 oocytes were fertilized (28%) and 516 of these (66%) were monospermic. This represents normal fertilization in 207 cycles (62% of total patient cycles). These 207 cycles produced an average of 2.6 +/- 1.8 embryos. The resultant 487 embryos were either transferred or cryopreserved for future transfer. A total of 42 pregnancies (21% per ET) resulted from 200 ET procedures with 397 embryos. Miscarriages or elective terminations have occurred (11/42 pregnancies, 26% to date) and 21 deliveries have resulted in 26 babies. There are nine ongoing pregnancies. Three of the pregnancies have been from cryopreserved embryos. Group B patients had the highest fertilization rate (23%) but the lowest pregnancy rate (7%). No other significant relationships between patient group and outcome were found. CONCLUSIONS: Subzonal insemination of multiple sperm is an effective treatment for some forms of male factor infertility.

Synthesis of (+/-)-[18F]BMY 14802, its enantiomers and their anatomical distributions in rodents.

A potential antipsychotic drug, BMY 14802 was labeled with 18F and its distribution in rodents was studied. No-carrier-added (NCA) (+/-)-[18F]BMY 14802 (5) was synthesized by two methods in 5-10% radiochemical yield in a synthesis time of 130-140 min from EOB with a specific activity of 0.5-1.5 Ci/microM. (+)- and (-)-[18F]BMY 14802 was synthesized by the chiral reduction of alpha-(4-[18F]fluorophenyl)-4-(5-fluoro-2-pyrimidinyl)-1-piperazine-b utanone (4) with chiral reducing agent, (+)- and (-)-beta-chlorodiisopinocampheylborane [(+)- and (-)-DIP chloride] in 6-10% radiochemical yield in a synthesis time of 150 min from EOB. Animal studies in mouse and in rat revealed that the distribution of 5 in each tissue was high at 5 min, the radioactivity then declined rapidly in all tissues studied except in the liver and in the small intestine. The radioactivity in the femur did not increase with time indicating in vivo defluorination may not occur. The uptakes of (+/-)-[18F]BMY 14802 and its enantiomers, (+)- and (-)-[18F]BMY 14802 in rat cerebellum, brain stem, hippocampus and spinal cord were similar and were significantly reduced by prior treatment of rat with haldol. This suggests that (+/-)-[18F]BMY 14802 and its enantiomers bind to sigma-receptors in a similar fashion.

Recent advances in the design and performance of research assistants.

Every laboratory needs personnel who work 24 hours a day 7 days a week, are efficient, obedient, capable of showing initiative, insightful and affable. Our experience of laboratory workers tells us that there is an immense way to go to achieve these goals. Having tired of the normal, Darwinian approach, a project has been initiated to achieve these ends through genetic technologies.

Comparative intracytoplasmic sperm injection (ICSI) in human and domestic species.

The current clinical use of intracytoplasmic sperm injection (ICSI) for the alleviation of male factor infertility has prompted a re-investigation of sperm injection techniques in a number of animal species. This report examines sperm injection of in vitro matured oocytes in the major domestic species and compares the results with the human. Ovine, bovine and porcine oocytes can undergo fertilization and at least limited development without exogenous activation either prior to or subsequent to injection. Porcine is temperature sensitive during fertilization and the early stages of embryo development. The oocytes of all three domestic species, particularly ovine, have a tendency to activate after the injection procedure regardless of the presence or absence of sperm. The implications for early development studies and the practical use of direct sperm injection for domestic species are discussed.

Sheep oocyte activation after intracytoplasmic sperm injection (ICSI).

The effect of calcium concentration on fertilization and activation was examined in oocytes injected in vitro with sperm. Oocytes were subjected to sperm injection, to sham injection or remained uninjected, and were then cultured for 19 h in bicarbonate-buffered synthetic oviduct fluid (BSOF) without calcium, or containing either calcium chloride or calcium ionophore. There was no difference in fertilization rates after ICSI when oocytes were cultured in vitro in media containing calcium chloride or calcium ionophore but the rate was lower in calcium-free media. There was also no difference in the fertilization rate after ICSI when oocytes were cultured in vivo compared with that observed in vitro in media containing calcium chloride or calcium ionophore. In calcium chloride-treated oocytes, activation was induced by mechanical injection, and in calcium ionophore-treated oocytes, by the ionophore. In uninjected oocytes, calcium itself did not cause oocyte activation. It is concluded that it is possible to induce activation by the injection process, but that manipulation alone is inadequate to cause proper oocyte activation unless calcium is also present. No difference in oocyte activation between ICSI and sham injection was found, indicating that the sperm may play no role in the early events of oocyte activation.

Injection of multiple sperm into the perivitelline space as a treatment of male infertility.

This paper reports the outcome of 274 treatment cycles using multiple injection of sperm into the perivitelline space as a treatment of male factor infertility. A total of 170 couples underwent this form of treatment; 59.1% of cycles had at least one oocyte normally fertilized with an overall normal fertilization rate of 17.2%. The development rate of normally fertilized embryos was high (98.5%) and resulted in a pregnancy rate (positive human chorionic gonadotrophin 18 days after embryo transfer) of 21.4% per embryo transfer procedure (a maximum of 3 embryos were transferred per procedure). The relationship between the number of sperm injected and the fertilization rate and other factors affecting the outcome are discussed.

Effect of culture, incubation and acrosome reaction of fresh and frozen-thawed ram spermatozoa for in vitro fertilization and intracytoplasmic sperm injection.

This study evaluated different sperm treatments for fertilization of sheep oocytes by intracytoplasmic sperm injection (ICSI) and in vitro fertilization (IVF). In Experiment 1, fresh and frozen semen was separated by Percoll centrifugation and incubated at 30 degrees C or 39 degrees C in HSOF or BSOF medium for 1 h before use for IVF or ICSI. For IVF, oocytes were inseminated and incubated with sperm for 30 min, 4 h and 19 h. Sperm were assessed for acrosome integrity after Percoll centrifugation and 1 h incubation, and those used for IVF were assessed after each period of exposure to the oocytes. Fertilization rates after ICSI were higher for fresh than for frozen-thawed sperm and were highest 19 h after IVF with fresh or frozen-thawed sperm in the presence of HSOF at 30 degrees C. In Experiment 2, fresh semen was separated by Percoll centrifugation and incubated for 5 h in HSOF, and the acrosome reaction was induced with lysophosphatidylcholine. Acrosome integrity was then assessed. Fertilization rates after ICSI were similar for acrosome-reacted and control spermatozoa. These results suggest that induction of the acrosome reaction in spermatozoa before ICSI is unnecessary, whereas a capacitating treatment of spermatozoa is required before IVF.

Clinical intracytoplasmic sperm injection (ICSI) results from Royal North Shore Hospital.

The technique of intracytoplasmic sperm injection (ICSI) was first introduced to the Royal North Shore Hospital in April 1993 as part of a controlled study of 100 patient cycles in which sibling oocytes were inseminated by either subzonal insemination (SUZI) or ICSI. This trial showed direct sperm injection to be superior in terms of fertilization. In that study, 58 embryo transfers of 101 ICSI-derived embryos resulted in 10 pregnancies. No miscarriages have occurred and a total of 10 fetal hearts (9.8% per embryo transferred) were detected on ultrasound. There have been 10 deliveries of 10 babies. Since the beginning of 1994, intracytoplasmic injection has been used exclusively for patients requiring micromanipulation to achieve fertilization. There have been 200 patient cycles with 1650 oocytes collected (8.8 oocytes per cycle). Of these oocytes, 1548 were mature (94%) and were subjected to ICSI, and normal fertilization occurred in 874 (56%) of the injected oocytes. The number of oocytes which cleaved and were suitable for fresh transfer or cryopreservation was 818 (94%). There have been 153 fresh embryo transfers of 326 embryos. Twenty-six pregnancies (17% per embryo transfer) have resulted, 22 of which proceeded to ultrasound examination in which 23 fetal hearts were detected (7% per embryo transferred). Three miscarriages have occurred, leaving 19 ongoing pregnancies. There have been 127 cryopreservation procedures involving 492 embryos. To date, there have been 47 embryo thaw cycles, and 93 of the 115 (81%) thawed embryos survived and were transferred. These 47 embryo transfers resulted in 10 pregnancies (21% per embryo transfer), one of which one has miscarried.(ABSTRACT TRUNCATED AT 250 WORDS)

Cleavage, development and competence of sheep embryos fertilized by intracytoplasmic sperm injection and in vitro fertilization.

More abnormal fertilization has been found in sheep oocytes after intracytoplasmic sperm injection (ICSI) than after in vitro fertilization (IVF). Although the birth of a normal lamb has been reported, the efficiency of blastocyst production is low. We therefore evaluated the cleavage, development and viability of sheep embryos obtained from ICSI, IVF and sham injection. In vitro matured oocytes either injected or inseminated with spermatozoa were assessed for cleavage 1 and 4 d after injection or insemination, and for development to blastocyst after 7 d of culture. A total of 699 oocytes was injected (ICSI); 198 (30.6%) were activated and 55 (8.5%) developed to the blastocyst stage. Of the 17 recipient ewes with 1, 2, 3 or 4 embryos, 15 (88.2%) were pregnant on Day 18; of these 17 recipients, 7 (41.1%) and 6 (35.2%) ewes remained pregnant on Days 45 and 110, respectively. Two normal lambs were born, one ewe died on Day 110 with 2 normal male fetuses, another ewe aborted on Day 90 and 4 pregnancies were maintained. A total of 517 oocytes was inseminated (IVF); 296 (62%) were activated and 90 (18.8%) reached the blastocyst stage. A total of 19 ewes received 1, 2, 3 or 4 embryos; of these, 13 (68.4%) were pregnant on Day 18, 8 (42.1%) ewes remained pregnant on each of Days 45 and 110. Three ewes delivered 5 lambs. Five pregnancies were maintained. A total of 156 oocytes was sham injected, 38 (24.3%) were activated and no blatocysts were obtained after culture. The results of this study showed that blastocysts obtained after ICSI are potentially viable and are not a result of parthenogenesis.

The selective removal of phosphorus from soil: is event size important?

Data from the Woburn Erosion Reference Experiment (Bedfordshire, UK) were used to test the hypothesis that losses of phosphorus (P) in small erosion events are as great as those in infrequent large events, and to examine the effect of storm characteristics on the selective enrichment of P in eroded sediment. For almost every plot event in the period 1988 to 1994, the clay-sized fraction of the sediment was enriched compared with the soil of the plots. There was more variation in clay enrichment for smaller erosion events than for larger ones. The clay and P contents of the sediment were strongly correlated (p < 0.01), and there was a wider range of P concentrations in the sediment derived from small events than in that from large events. However, individual events resulting in small soil losses (< 100 kg) did not account for greater P losses than larger events (> 100 kg). The greater frequency of smaller events, combined with the likelihood of higher P concentrations in the sediment, therefore accounted for a greater proportion of the P lost over the 6-yr period than the infrequent large events. Phosphorus concentrations generally increased with increasing peak discharge and decreased with increasing event duration. For the same return period, P losses were generally greater from plots cultivated up and down the slope than from those cultivated across the slope. Overall, our results suggest that small erosion events should be controlled to prevent P contamination of surface waters and that the most effective means of doing this are by the introduction of minimal tillage techniques and across-slope cultivations.

Birth of a male lamb derived from an in vitro matured oocyte fertilised by intracytoplasmic injection of a single presumptive male sperm.

The developmental competence of in vitro matured ovine oocytes, cytoplasmically injected with single male or female chromosome-bearing sperm, was investigated. Eighty-five unsorted, 92 'female-sorted' and 74 'male-sorted' ram sperm were injected into in vitro matured sheep oocytes and, two to four hours later, placed into the oviducts of 28 oestrous sheep. The sperm were separated according to sex by analysis of their DNA content with a flow cytometer. One pregnancy was diagnosed by ultrasound after 55 days and a 3 kg male lamb was born after 150 days gestation. This lamb was derived from an oocyte injected with 'male-sorted' sperm.