The efficacy of cefazolin plus macrolide (erythromycin or clarithromycin) versus cefazolin alone in neonatal morbidity and placental inflammation for women with preterm premature rupture of membranes.

INTRODUCTION: Although the use of broad-spectrum antibiotics in women with preterm premature rupture of membranes (PPROM) is recommended to prolong pregnancy and decrease short-term neonatal complications, the optimal regimen remains undetermined. The objective of this study was to compare the efficacy of cefazolin plus macrolide (erythromycin or clarithromycin) versus cefazolin alone in reducing neonatal morbidity and placental inflammation for women with PPROM. METHODS: This prospective study included singleton pregnancies with PPROM (23-33 weeks gestation). The primary outcome was neonatal composite morbidity and the secondary outcomes were the incidence of abnormal brain sonography and infant neurological outcome at one year of age. The presence and the stage of acute histological chorioamnionitis and funisitis were also reviewed blinded to all clinical information. RESULTS: 102 women were randomly assigned to cefazolin (n = 35), cefazolin plus erythromycin (n = 31), or cefazolin plus clarithromycin (n = 36). The neonatal composite morbidity, the incidence of abnormal brain sonography, and infant neurological outcome at one year of age were similar between the comparison treatments (combination of cefazolin plus erythromycin or clarithromycin) and cefazolin. However, the presence and stage of histological funisitis showed significant difference between cefazolin plus clarithromycin versus cefazolin alone (p = 0.023). DISCUSSION: This study is the first clinical trial of the use of cefazolin with either clarithromycin or erythromycin compared to cefazolin alone in the management of PPROM in which the primary and secondary analyses showed no difference among the three antibiotic regimens. The only noted difference was from a lesser degree of histological funisitis associated with clarithromycin exposure. CONCLUSION: Our data suggests that clarithromycin may be an alternative worth considering with potentially beneficial effects compared to erythromycin in PPROM.

Exogenous DNA uptake of boar spermatozoa by a magnetic nanoparticle vector system.

The sperm-mediated gene transfer method is applicable to transgenesis in many species that use spermatozoa for reproduction recently, which has been shown various results. In the current study, we show that transgenic porcine embryos can be efficiently produced by employing a simple transfection method that uses magnetic nanoparticles (MNPs). The complexes formed between plasmid DNA and MNPs were bounded on ejaculated boar spermatozoa at a higher efficiency compared to methods using DNA alone or lipofection. Using confocal microscopy, rhodamine fluorophore-labelled MNPs were detected on external surfaces of the spermatozoa membrane, which were bounded on zona pellucida of in vitro maturated oocyte during in vitro fertilization. Electron microscopy revealed that clusters of MNPs were detected in inside of plasma membrane and nucleus of the spermatozoa head. Additionally, we found that magnetofected boar spermatozoa could be fertilized with oocytes in vitro and that the resulting gene of green fluorescent protein was detected in fertilized eggs by genomic PCR analysis. Taken together, these results suggest that MNPs can be used to efficiently introduce a transgene into embryo via spermatozoa.

Stimulation of plasminogen activator activity by free radicals in boar spermatozoa.

Plasminogen activators (PAs), commonly found on the membrane of spermatozoa, convert plasminogen into plasmin and may participate in mammalian fertilization. Correlations have been reported between reactive oxygen species (ROS) and spermatozoa function, although the relationship between PA activity and ROS is unknown. We investigated the effects of ROS on PA activity. We used an in vitro model of free radical generation whereby boar spermatozoa were preincubated in xanthine and xanthine oxidase (X-XO) and PA activity was then measured. The acrosome reaction of boar spermatozoa was significantly promoted by 100 mU/mL plasmin (P<0.01), similar to levels achieved when stimulated with the positive calcium (2 mM) control. The addition of plasminogen to the fertilization medium significantly promoted both spermatozoa binding (157.5+/-14.0 spermatozoa/oocyte) and the percentage of oocytes with a male pronucleus (74.5+/-6.4%) compared with control (98.4+/-21.8 spermatozoa/oocyte and 51.4+/-5.3%, respectively; P<0.05). The acrosome reactions of spermatozoa were significantly higher when incubated with calcium (2 mM; 60.2+/-2.7%), calcium (2 mM)+EDTA (6 mM; 29.4+/-4.2%), sodium nitroprusside (0.1 microM; 38.0+/-4.2%), H(2)O(2) (100 microM; 56.0+/-3.0%), and X-XO (0.5 mM and 0.05 U/mL, respectively; 31.8+/-3.7%) compared with non-capacitation medium as control (19.0+/-2.7%; P<0.05). However, when spermatozoa were incubated with only X-XO, PA activity was significantly higher than with other treatments (P<0.05). Moreover, the addition of the antioxidant superoxide dismutase to the X-XO system significantly blocked the PA activity of spermatozoa (P<0.05). The PA activity of spermatozoa treated with X-XO was significantly reduced by the addition of MEK inhibitor (55.2+/-5.6 ng/mL) and p38 inhibitor (57.4+/-2.7 ng/mL), but not PI3K inhibitor, compared to the control (X-XO; 68.0+/-5.8 ng/mL; P<0.05). The induction of PA activity in boar spermatozoa by free radical generation suggests the PA/plasmin system plays a role in mammalian fertilization.

Phenotypic evolution of classic 21-hydroxylase deficiency.

We describe a female patient who was diagnosed and treated at birth for a classic form of salt-losing congenital adrenal hyperplasia. At 17 years of age, against medical advice, she discontinued both mineralocorticoid and glucocorticoid replacement with no resulting clinical symptoms other than the occurrence of amenorrhoea. Steroid metabolites revealed significant abnormalities of the renin-angiotensin-aldosterone axis, as well as of pituitary-adrenal function. Analysis of our patient's DNA showed only one deleterious CYP21 mutation, an intron 2 base pair change activating a cryptic splice site. We speculate that expression of this patient's CYP21 genes may be altered by the effects of ageing or by changes in the steroid milieu.