Percutaneous umbilical blood sampling and umbilical vein transfusions. Rapid serologic differentiation of fetal blood from maternal blood

Percutaneous umbilical blood samples (PUBS), obtained under ultrasound guidance, are used for prenatal diagnosis and management of hemolytic disease of the newborn (HDN) and other fetal disorders. Rapid testing at the time of sampling is vital to distinguish fetal from maternal blood. Blood typing was performed by slide technique in the treatment room during 38 procedures on 25 patients. Anti-I was used to test 50 presumed PUBS; venous I-positive maternal blood was tested in parallel. Because anti-I cannot detect fetal blood after umbilical vein transfusion (UVT) of I-positive donor blood, ABO and Rh blood typing reagents were used to test 29 samples when maternal and fetal or donor blood groups differed. Monoclonal reagents were used for optimal detection of weak AB antigens in fetal blood. Avid, chemically modified anti-D was used for Rh typing. Blood typing showed 27 (34%) of 79 samples to be maternal blood. Fetal blood was obtained in 8 of 10 cases investigated for fetal disorder and in 16 cases of potential HDN (anti-D, 5; -CD, 5; -cE, 2; -K, 2; -c; -E). The absence of HDN (antigen-negative fetus) was determined in 4 cases. UVT afforded live birth of 9 of 10 infants with HDN and was not indicated in two cases.     

Radioiodinated dechloro-4-iodofenofibrate: a hydrophobic model drug for molecular imaging studies

The aptitude of ropinirole to permeate the buccal tissue was tested using porcine mucosa mounted on Franz-type diffusion cells as ex vivo model. Drug permeation was also evaluated in presence of various penetration enhancers and in iontophoretic conditions. Ropinirole, widely used in treatment of motor fluctuations of Parkinson's disease, passes the buccal mucosa. Flux and permeability coefficient values suggested that the membrane does not appear a limiting step to the drug absorption. Nevertheless, an initial lag time is observed but the input rate can be modulated by permeation enhancement using limonene or by application of electric fields. Absorption improvement was accompanied by the important reduction of the lag time; at once the time required to reach the steady state plasma concentration was drastically decreased. On the basis of these results we could assume that clinical application of ropinirole by buccal delivery is feasible.     

Reduction of Renal Superoxide Dismutase in Progressive Diabetic Nephropathy

Superoxide excess plays a central role in tissue damage that results from diabetes, but the mechanisms of superoxide overproduction in diabetic nephropathy (DN) are incompletely understood. In the present study, we investigated the enzyme superoxide dismutase (SOD), a major defender against superoxide, in the kidneys during the development of murine DN. We assessed SOD activity and the expression of SOD isoforms in the kidneys of two diabetic mouse models (C57BL/6-Akita and KK/Ta-Akita) that exhibit comparable levels of hyperglycemia but different susceptibility to DN. We observed down-regulation of cytosolic CuZn-SOD (SOD1) and extracellular CuZn-SOD (SOD3), but not mitochondrial Mn-SOD (SOD2), in the kidney of KK/Ta-Akita mice which exhibit progressive DN. In contrast, we did not detect a change in renal SOD expression in DN-resistant C57BL/6-Akita mice. Consistent with these findings, there was a significant reduction in total SOD activity in the kidney of KK/Ta-Akita mice compared with C57BL/6-Akita mice. Finally, treatment of KK/Ta-Akita mice with a SOD mimetic, tempol, ameliorated the nephropathic changes in KK/Ta-Akita mice without altering the level of hyperglycemia. Collectively, these results indicate that down-regulation of renal SOD1 and SOD3 may play a key role in the pathogenesis of DN.Diabetic nephropathy (DN) is the leading cause of end-stage renal disease. Although hyperglycemia is clearly a prerequisite for the development of DN, alone it is insufficient for its development. Epidemiologic studies demonstrate only 10% to 40% of all diabetic patients get DN, despite comparable levels of glucose control in those subjects developing DN versus spared. In addition, sibling studies show a strong familial component for the risk of developing persistent proteinuria, suggesting a genetic basis for DN risk.^1,2 However, the molecular or cellular mechanisms coupled with the genetic susceptibility to DN are incompletely understood.There is compelling evidence that superoxide excess induced by diabetic hyperglycemia plays a central role in diabetic vascular cell damage.³ High glucose flux increases the production of superoxide anion (O₂^•−) by mitochondrial electron-transport chain, and the overproduced superoxide enhances the major pathways of hyperglycemic vascular celldamage, including protein kinase C, advanced glycation end (AGE) products, and hexosamine pathways.⁴ In addition, superoxide is produced by multiple pathogenic pathways of diabetes. These include increased nicotinamide adenine dinucleotide phosphate [NAD(P)H] oxidase activity, uncoupled endothelial nitric oxide synthase (eNOS), and enhanced signaling of AGEs, angiotensin II, and oxidized-LDL receptors.⁵ Excessive production of superoxide anion results in the formation of secondary reactive oxygen species (ROS) including peroxynitrite and hydroxyl radicals, leading the damage of DNA, proteins, and lipids, and causes vascular cell injury.⁶ Thus, superoxide overproduction is considered as a major pathogenic pathway in diabetic vascular complication.A net accumulation of superoxide anion is determined by a balance between superoxide production and antioxidant capacity. In this context, antioxidant defense system could play a critical role in diabetic vascular damage. Superoxide dismutase (SOD) is the major antioxidant enzyme for superoxide removal, which converts superoxide into hydrogen peroxide (H₂O₂) and molecular oxygen.^7,8 The hydrogen peroxide is further detoxified to water (H₂O) by catalase or glutathione peroxidase.^6,9 In mammals, three SOD isoforms exist: cytoplasmic CuZnSOD (SOD1), mitochondrial MnSOD (SOD2), and extracellular CuZnSOD (SOD3, ecSOD).^8,10 Each SOD isoform is derived from distinct genes but catalyzes the same reaction, producing H₂O₂ from O₂^•−.¹⁰ There is substantial evidence that SOD activity in peripheral blood cells is reduced in the diabetic patients with DN as compared with those without diabetic complication.^11–14 In addition, recent studies have implicated SOD (SOD1 and SOD2) gene polymorphism in human DN risk.^15–17 Furthermore, it was shown that the transgenic mice with SOD (SOD1 or SOD2) gene are resistant to diabetes-induced vascular injuries, including nephropathy.^4,18,19 In aggregate, these findings suggest a pivotal role of SOD enzyme in the pathogenesis of DN. However, the changes in renal SOD enzymes in DN and their significance are poorly described.Recent studies of streptozotocin (STZ)-induced diabetic mice have shown that genetic factors significantly affect the development and the severity of DN in mice as well as in human.²⁰ Among the inbred strains of mice, KK/HlJ and DBA/2 strains have been identified as DN-prone strains, whereas the widely used C57BL/6 strain is relatively resistant to DN.²¹ Compared with the STZ model, spontaneously diabetic mice offer a unique opportunity to assess the pathogenic pathways in DN without the potential nonspecific tissue toxicity of STZ. Ins2^Akita mouse (Akita mouse) is a well-studied nonobese hypoinsulinemic diabetic mouse.^22,23 This diabetic strain has a mutation in cysteine 96 to tyrosin in the insulin 2 gene (Akita mutation) and exhibits marked hyperglycemia as early as 4 wk of age.²³ However, Akita mouse does not develop overt DN due to C57BL/6 background.²⁴ To investigate renal alteration of SOD enzyme in advanced DN without the nonspecific tissue toxicity of STZ, we here generated a new congenic strain of the Akita mutation that exhibits progressive DN by backcrossing C57BL/6-strain Akita mouse (C57BL/6-Akita) to the nephropathy prone KK/Ta strain mouse.²⁵ Our data demonstrate that renal expression of SOD1 and SOD3, but not SOD2, is prominently down-regulated in KK/Ta-strain Akita mouse (KK/Ta-Akita), which exhibits progressive DN, whereas renal SOD expression was not altered in the DN-resistant C57BL/6-Akita mouse. Furthermore, the present study demonstrates that treatment with tempol, a SOD mimetic, remarkably ameliorates the nephropathic changes in KK/Ta-Akita mice. Taken together, these results suggest that downregulation of renal SOD1 and SOD3 may play a key role in the pathogenesis of DN.     

Associations among hypospadias,cryptorchidism, anogenital distance,and endocrine disruption

Endocrine disruptors, such as environmental compounds with endocrine-altering properties, may cause hypospadias and crytorchidism in several species, including humans. Anogenital distance is sexually dimorphic in many mammals, with males having longer anogenital distance on average than females. Animal models of proposed endocrine disruptors have associated prenatal exposure with hypospadias, cryptorchidism, and reduced anogenital distance. Human studies have correlated shorter anogenital distance to in utero exposure to putative endocrine disruptors. We review preliminary data suggesting that anogenital distance is reduced in boys with hypospadia and cryptorchidism. Hence, human hypospadias and cryptorchidism may be associated with reduced anogenital distance as a result of endocrine disruption.     

Physiologic and pathophysiologic roles of lipid mediators in the kidney

Small lipids such as eicosanoids exert diverse and complex functions. In addition to their role in regulating normal kidney function, these lipids also play important roles in the pathogenesis of kidney diseases. Cyclooxygenase (COX)-derived prostanoids play important role in maintaining renal function, body fluid homeostasis, and blood pressure. Renal cortical COX2-derived prostanoids, particularly (PGI2) and PGE2 play critical roles in maintaining blood pressure and renal function in volume contracted states. Renal medullary COX2-derived prostanoids appear to have antihypertensive effect in individuals challenged with a high salt diet. 5-Lipoxygenase (LO)-derived leukotrienes are involved in inflammatory glomerular injury. LO product 12-hydroxyeicosatetraenoic acid (12-HETE) is associated with pathogenesis of hypertension, and may mediate angiotensin II and TGFbeta induced mesengial cell abnormality in diabetic nephropathy. P450 hydroxylase-derived 20-HETE is a potent vasoconstrictor and is involved in the pathogenesis of hypertension. P450 epoxygenase derived epoxyeicosatrienoic acids (EETs) have vasodilator and natriuretic effect. Blockade of EET formation is associated with salt-sensitive hypertension. Ceramide has also been demonstrated to be an important signaling molecule, which is involved in pathogenesis of acute kidney injury caused by ischemia/reperfusion, and toxic insults. Those pathways should provide fruitful targets for intervention in the pharmacologic treatment of renal disease.     

Haplotype analysis of CYP11A1 identifies promoter variants associated with breast cancer risk

The CYP11A1 gene encodes the cholesterol side chain cleavage enzyme that catalyzes the initial and rate-limiting step of steroidogenesis. A large number of epidemiologic studies have implicated the duration and degree of endogenous estrogen exposure in the development of breast cancer in women. Here, we conduct a systematic investigation of the role of genetic variation of the CYP11A1 gene in breast cancer risk in a study of 1193 breast cancer cases and 1310 matched controls from the Shanghai Breast Cancer Study. We characterize the genetic architecture of the CYP11A1 gene in a Chinese study population. We then genotype tagging polymorphisms to capture common variation at the locus for tests of association. Variants designating a haplotype encompassing the gene promoter are significantly associated with both increased expression (P = 1.6e-6) and increased breast cancer risk: heterozygote age-adjusted odds ratio (OR), 1.51 [95% confidence interval (95% CI), 1.19-1.91]; homozygote age-adjusted OR, 2.94 (95% CI, 1.22-7.12), test for trend, P = 5.0e-5. Among genes controlling endogenous estrogen metabolism, CYP11A1 harbors common variants that may influence expression to significantly modify risk of breast cancer.     

骨髓基质干细胞移植对正常和心肌梗死大鼠心电图及心功能的影响

目的:骨髓基质干细胞移植到心肌梗死的瘢痕心肌组织中可以改善心功能,但以心电图为观察指标的研究不多。实验观察骨髓基质干细胞移植对正常和心肌梗死大鼠心电图及心功能的影响。方法:实验于2004-01/2005-03在哈尔滨医科大学完成。①实验动物:选取4周龄雄性Wistar大鼠80只,随机数字表法分为梗死移植组、正常移植组、梗死非移植组、正常非移植组,20只/组。另选取7d龄Wistar雄鼠30只作为骨髓基质干细胞的来源。②实验方法:采用密度梯度离心法获取鼠骨髓基质干细胞,配成1×109L-1的细胞悬液,使用5-氮胞苷体外诱导培养3～4周,移植前24～48h行Brdu标记。取载有细胞的盖玻片,测定钙释放时将20mmol/L的caffeine快速加在细胞表面。梗死移植组、梗死非移植组大鼠建立心肌梗死模型。造模4周后,梗死移植组将0.25mL诱导的骨髓基质干细胞悬液注射至大鼠心肌梗死后的瘢痕组织,正常移植组同法将骨髓基质干细胞悬液注射至正常心肌组织,梗死非移植组、正常非移植组注射等量不含骨髓基质干细胞的培养液基质。③实验评估:观察骨髓基质干细胞的诱导分化情况及其植入后在瘢痕心肌组织中的生存状态。测定细胞内钙离子浓度。记录术前、冠脉结扎后即刻/细胞移植即刻、术后4周的心电图变化。检测术后4周的超声和血流动力学指标变化。结果:80只大鼠均进入结果分析。①骨髓基质干细胞的诱导分化及其植入后的生存状态:5-氮胞苷诱导3周后,骨髓基质干细胞表达肌钙蛋白Ⅰ和肌凝蛋白重链,细胞内有丰富的肌丝和Z线,细胞器较多。植入4周后在心肌瘢痕组织中分化为心肌细胞。②细胞内钙离子浓度:两组细胞在caffeine刺激下钙离子的释放均呈波峰状,但诱导组应用caffeine后钙离子浓度降低且低于基础状态,钙释放受到抑制,未诱导组不受影响。③心电图观察:与术前比较,梗死移植组QRS波变窄,R波降支出现正常顿挫波,未见显著心律失常。④超声检测及血流动力学分析:术后4周,与梗死非移植组比较,梗死移植组左室收缩末压、左室射血分数和压力变化速率最大值均显著升高(P<0.05或0.01)。结论:骨髓基质干细胞体外诱导后能分化为心肌样细胞,植入到瘢痕心肌组织中生存、增殖良好,可改善心电图及心肌弹性,从而改善心肌梗死大鼠的心功能。     

Sexual dysfunction, HIV, and AIDS in men who have sex with men

HIV infection is associated with sexual dysfunction. Using validated instruments, we investigated the relationship between HIV/AIDS and sexual function in a contemporary cohort of men who have sex with men (MSM). An anonymous Internet-based survey was disseminated to MSM via organizations and social networking sites that cater to this population. Information on ethnodemographic variables, health status (including HIV status, disease stage, and other health conditions), and sexual behavior was collected. Men were categorized as HIV-negative, HIV-positive/AIDS-negative, or HIV-positive /AIDS-positive. A modified validated version of the International Index of Erectile Function (IIEF) for use in MSM and the Premature Ejaculation Diagnostic Tool (PEDT) were used to stratify risk of sexual dysfunction. The study cohort included 1361 men (236 of whom were HIV-positive) who provided complete data on HIV status, IIEF, and PEDT. There was a significant trend toward greater prevalence of erectile dysfunction (ED) in men with progressive HIV infection 40-59 years of age relative to age matched HIV-negative men (p=0.02). In a logistic regression model controlling for other variables, HIV infection without AIDS was not associated with greater odds of ED; however, HIV infection with AIDS was associated with greater odds of ED (p=0.006). In a separate logistic regression model, HIV infection with or without AIDS was not significantly associated with greater odds of premature ejaculation (p>0.05). Use of phosphodiesterase 5 (PDE5) inhibitor drugs was much more common in HIV-infected men. HIV infection is a risk factor for poorer sexual function primarily due to higher risk of erectile dysfunction in men with AIDS.