Selection of birefringent spermatozoa under Polscope: effect on intracytoplasmic sperm injection outcome.

The ideal method for sperm selection during Intracytoplasmic sperm injection (ICSI) is still ill-defined. Identification of a viable spermatozoon amongst immotile spermatozoa for ICSI often becomes difficult. Ninety-six ICSI cycles were selected and divided into Group A (azoospermic, n = 58) and Group B (complete asthenozoospermic, n = 38). Oocytes having birefringent meiotic spindle and zona pellucida thickness <20 µm were selected for ICSI. Groups A and B were further divided into A1, A2 and B1, B2, respectively, based on the type of ICSI performed. In Group A1, a motile spermatozoon with normal morphology was injected into a metaphase-II (M-II) oocyte. In Group B1, spermatozoon showing coiling of tail following modified hypo-osmotic swelling test was injected into M-II oocytes. In Groups A2 and B2, ICSI was performed by injecting a spermatozoan with birefringent head. Pronuclear morphology, fertilisation rate, embryo grading and pregnancy rate were assessed. ICSI outcome measures were better in Group A2 than in Group A1 but were statistically insignificant. However, significantly higher percentage of Z1 and Z2 zygotes, Grade I and Grade II embryos and pregnancy rate were observed in Group B2 as compared to Group B1. Selection of birefringent spermatozoa shows promising results in asthenozoospermic men and men undergoing testicular sperm aspiration or extraction before ICSI.

Safety of outpatient treatment in acute pulmonary embolism.

INTRODUCTION: Data regarding outpatient treatment of pulmonary embolism (PE) is scarce. This study evaluates the safety of outpatient management of acute PE. METHODS: This is a retrospective cohort study of consecutive patients presenting at the Ottawa Hospital with acute PE diagnosed between 1 January 2007 and 31 December 2008. PE was defined as an arterial filling defect on CTPA or a high probability V/Q scan. Patients were managed as outpatients if they were hemodynamically stable, did not require supplemental oxygenation and did not have contraindications to low-molecular-weight heparin therapy. RESULTS: In this cohort of 473 patients with acute PE, 260 (55.0%) were treated as outpatients and 213 (45.0%) were admitted to the hospital. The majority of the patients were admitted because of severe comorbidities (45.5%) or hypoxia (22.1%). No outpatient died of fatal PE during the 3-month follow-up period. At the end of follow-up, the overall mortality was 5.0% (95% CI, 2.7-8.4%). The rates of recurrent venous thromboembolism (VTE) in outpatients were 0.4% (95% CI, 0.0-2.1%) and 3.8% (95% CI, 1.9-7.0%) within 14 days and 3 months, respectively. The rates of major bleeding episodes were 0% (95% CI, 0-1.4%) and 1.5% (95% CI, 0.4-3.9%) within 14 days and 3 months, respectively. Four (1.5%) outpatients were admitted to the hospital within 14 days. CONCLUSIONS: A majority of patients with acute PE can be managed as outpatients with a low risk of mortality, recurrent VTE and major bleeding episodes.

Laparoscopic repair of two unusual traumatic colorectal injuries.

Although the role of laparoscopy in the repair of iatrogenic colonic perforations in prepared bowel is well established, the use of laparoscopy as a therapeutic tool in the management of patients with traumatic intraabdominal injuries remains unclear. We report two cases of traumatic colorectal injuries that were managed successfully by therapeutic laparoscopy. Both patients sustained traumatic perforations to the large bowel as a result of unusual accidents in the home. Laparoscopic repair of the perforations with interrupted sutures and peritoneal lavage were performed with good results in both cases. We believe that the laparoscopic repair of traumatic colonic injuries is feasible in selected patients and offers a number of accepted advantages over formal laparotomy.

Microplate enzyme-linked immunosorbent assay for the detection of primary DNA alterations based on the interaction with UvrA/UvrB.

An enzyme-linked microplate immunoassay for the analysis of primary DNA lesions is described. The assay principle is based on the interaction of the bacterial DNA repair proteins UvrA and UvrB with DNA and on the immunodetection of UvrB forming a stable complex with covalently modified nucleotides. Using this technique we were able to detect damages in genomic DNA induced by uv light and by several different genotoxic agents. The detection sensitivity of the method reaches down to the nanomolar range of the mutagenic compound depending on the type of the DNA alteration. The method might be used in automated high-throughput studies.

Regioselective bromination of organic substrates by tetrabutylammonium bromide promoted by V2O5-H2O2: an environmentally favorable synthetic protocol

[reaction: see text] Vanadium pentoxide very effectively promotes the bromination of organic substrates, including selective bromination of some aromatics, by tetrabutylammonium bromide in the presence of hydrogen peroxide; mild conditions, high selectivity, yield, and reaction rate, and redundancy of bromine and hydrobromic acid are some of the major advantages of the synthetic protocol.

Cytokine depot formulations as adjuvants for tumor vaccines. I. Liposome-encapsulated IL-2 as a depot formulation.

In an attempt to mimic cytokine gene-transfected tumor cells and to develop an alternative approach to cancer immunotherapy, the authors vaccinated mice with mixtures of inactivated tumor cells and cytokine-containing depots. The RenCa mouse renal carcinoma and the B16 mouse melanoma were used as animal tumor models, with interleukin-2 (IL-2) as a cytokine and liposomes as a depot form. The results obtained show that vaccines consisting of mixtures of irradiated tumor cells and cytokine-containing liposomes can be used as highly effective tumor vaccines. These vaccines are very easy to prepare and, in contrast to vaccines consisting of cytokine gene transfected tumor cells, their composition (cell dosage, cytokine dosage) can be easily varied. Vaccination efficiency depended on (a) on the immunogenicity of the tumor cells: RenCa tumor cells are more immunogenic than B16 melanoma cells; (b) vaccination frequency: a single vaccination with irradiated tumor cells and 10 micrograms of IL-2 in liposome-encapsulated form was sufficient to induce lasting protective immunity against the RenCa tumor, whereas several (four to six) vaccinations in weekly intervals were needed to obtain a similar degree of protective immunity to the B16 melanoma; and (c) the dose of the cytokine encapsulated in the admixed liposome depots: immunity to the tumors could be induced only within a narrow cytokine-dose range ("IL-2-dose window"). The results obtained indicate that, because of the easiness of preparation and handling, vaccine formulations consisting of irradiated tumor cells and IL-2 in depot formulations are candidates for tumor vaccines for the treatment of tumor patients.

The isoprenoid pathway: cloning and characterization of fungal FPPS genes.

Farnesylpyrophosphate synthase (FPPS) is a key enzyme in isoprenoid biosynthesis. Several classes of essential metabolites, including sterols, quinones, carotenoids and gibberellins, are terpenoids with high biological activity. The structural gene for FPP synthase was isolated from two ascomycete fungi, Neurospora crassa and Gibberella fujikuroi. A comparative analysis of the nucleotide sequences of both FPPS genes revealed the presence of introns at the same positions at the 5' end of the coding regions. Furthermore, the most conserved region of the gene was isolated from two other plant pathogenic fungi, Sphaceloma manihoticola and Claviceps purpurea, by PCR. Sequence analysis showed a high degree of similarity between the deduced proteins of all known FPP synthase genes. In contrast to animals, all analyzed fungi contain a single copy of the gene, although FPP is the precursor for essential sterol and quinone biosynthesis and secondary metabolites, such as gibberellins, as well. Transcription analysis in different light regimes has shown that the FPPS genes in G. fujikuroi and N. crassa are not regulated by light induction.

Trial of vitamin A supplementation in very low birth weight infants at risk for bronchopulmonary dysplasia.

We performed a randomized, double-blind, controlled trial to determine whether vitamin A supplementation in a group of very low birth weight infants would reduce the incidence of bronchopulmonary dysplasia. Forty-nine infants (birth weight 700 to 1100 gm) requiring mechanical ventilation and supplemental oxygen at 96 hours age were randomly assigned to receive either 2000 IU retinyl palmitate (n = 27) or saline placebo (n = 22) intramuscularly every other day for up to 14 doses. There were no differences between treatment groups in the incidences of bronchopulmonary dysplasia at 31 days of postnatal age (vitamin A group 48%, placebo group 55%; p = 0.776), supplemental oxygen requirement at 34 weeks of postconceptional age, or other complications of prematurity. The vitamin A group had higher mean plasma vitamin A concentrations than the placebo group, but mean plasma vitamin A concentrations were greater than 20 micrograms/dl (suggesting sufficiency) in both groups after the first study week. By study day 28, only one fourth of the infants in either group had plasma vitamin A concentrations less than 20 micrograms/dl. In contrast to an earlier report, we found no change in the incidence of BPD with vitamin A supplementation. Our findings may reflect a low baseline incidence of vitamin A deficiency in the study population and recent changes in the respiratory care of very low birth weight infants. The latter may have lessened the potential impact of vitamin A deficiency on lung disease.

Results of laboratory surveillance of very low birth weight infants treated with a synthetic surfactant.

During two randomized, controlled trials comparing single prophylactic doses of air placebo and a synthetic surfactant, Exosurf Neonatal, many routine laboratory measurements were made in small premature infants during the first week of life. Values for serum electrolytes, urea, creatinine, phospholipid, cholesterol, glucose, calcium, bilirubin, and liver enzymes, as well as for hemoglobin, leukocyte and platelet counts, and urinalysis, did not differ between the placebo- and surfactant-treated infants. There was no evidence that prophylactic administration of this synthetic surfactant causes alterations in standard laboratory tests in small premature infants.

Improved outcome at 28 days of age for very low birth weight infants treated with a single dose of a synthetic surfactant.

Two identical double-blind, controlled, randomized trials were initiated to determine whether the administration of a single 5 ml/kg dose of a synthetic surfactant (Exosurf Neonatal), soon after the delivery of infants with birth weights 700 to 1350 gm, would improve rates of survival without bronchopulmonary dysplasia. Both trials were terminated before enrolling their planned sample sizes because of the availability of Exosurf under the provisions of a Treatment Investigational New Drug program. We report the combined results of these trials. Study infants were stratified according to birth weight and gender before random assignment to a treatment regimen. One hundred ninety-two infants received Exosurf and 193 received an air placebo. The study groups were similar when a variety of demographic features describing the mothers, their pregnancies, the circumstances of the births, and the infants were compared. Exosurf-treated infants required significantly less oxygen and respiratory support during the first 3 days of life in comparison with the air-treated infants. Fewer infants in the Exosurf group had pulmonary interstitial emphysema (26 vs 13; p = 0.028). In the Exosurf group, there was a significant reduction in the combined outcome, neonatal death or survival with bronchopulmonary dysplasia (57 vs 39; p = 0.042), and there was a significant increase in rates of survival without this disease (128 vs 137; p = 0.042). There were no differences between treatment groups in the incidences of a variety of complications of prematurity, including apnea, patent ductus arteriosus, intraventricular hemorrhage, and necrotizing enterocolitis. We conclude that improvements in respiratory physiology after a single prophylactic dose of Exosurf result in an increased likelihood of neonatal survival without bronchopulmonary dysplasia.

Pulmonary function following positive pressure ventilation initiated immediately after birth in infants with respiratory distress syndrome.

Cardiopulmonary function following positive pressure ventilation, initiated immediately after birth, was evaluated in 10 very low birthweight infants with respiratory distress syndrome (RDS; RDS infants). Multiple gas rebreathing methodology was used to measure functional residual capacity (FRC), diffusing capacity of the lung for carbon monoxide (DLCO) and effective pulmonary capillary blood flow (Qeff) at 2, 24, and 72 hr of age. Cardiopulmonary function variables were also measured at 2 hr of age in 10 infants of similar birthweight who did not have RDS (non-RDS infants). In RDS infants, mean FRC at both 2 hr of age (15 mL/kg or 0.42 mL/cm) and 24 hours of age (20 mL/kg or 0.54 mL/cm) was less than published predicted values for healthy infants and significantly less than values in non-RDS infants at 2 hours of age (29 mL/kg or 0.8 mL/cm). By 72 hr of age, mean FRC in RDS infants rose to predicted. At 2 and 24 hours of age, mean Qeff in RDS infants (41 and 38 mL/min/kg, respectively) was below predicted, as well as below the value observed in non-RDS infants at 2 hr of age (62 mL/min/kg). in RDS infants. DLCO remained well below published predicted values throughout the first three days of life. We conclude that early aggressive respiratory therapy does not result in the establishment of normal lung volumes during the first day of life, but it does so by 3 days of age. However, in spite of intubation immediately after birth and the application of continuous positive pressure ventilation during the first 3 days of life, very low birthweight infants with RDS continue to have significant alteration in lung function, evidenced by impaired diffusing capacity of the lung and low arterial-alveolar oxygen tension ratios.