7 P De Togn e t a / , i b~d , p 703, T. A Banks et a1 , J /n?n?uno/ 155, 1685 (1 995) 8 Mce were ~mmun~zed intrapertoneally w~th 100 &I of phosphate-buffered sane contanng 10% SRBCs (22) Ten days later, spleens were harvested, embedded In 0 C.T compound (M~les), and frozen In q u d ntrogen Frozen t~ssue sections (6 to 10 &m th~ck) were f~xed n cold acetone Endogenous per-ox~dase was q~~enched w~th 0 2% H, O, n methanol Sectons were sta~ned by f~rst ncubatng w~t h PNA-b~o t~n (Vector) and w~th rat antserum produced to IgD (Southern B~otechnology) After wash~ng, the sections were further incubated w~th strepta\/id~n conj~lgated w~th alkalne phosphatase (AP) (Zymed) and rabb~t ant~body to rat IgG conjugated w~th horse-rad~sh perox~dase (HRP) (Southern B~otechnology) Color development for bound AP and HRP was w~th an AP reacton k~t (Vector) and w~t h d~arn~nobenz-d~ne Sect~ons were then countersta~ned w~th 1 % methyl green 9 M. L Rose, M S C B~rbeck, V J Wall~s, J A Forrester. A J Daves. Nature 284, 364 (1980) 10 T

Mechanical overload may change cardiac structure through angiotensin II-dependent and angiotensin IIindependent mechanisms. We investigated the effects of mechanical strain on the gene expression of tenascin-C, a prominent extracellular molecule in actively remodeling tissues, in neonatal rat cardiac myocytes. Mechanical strain induced tenascin-C mRNA (3.9 ؎ 0.5-fold, p < 0.01, n ‫؍‬ 13) and tenascin-C protein in an amplitude-dependent manner but did not induce secreted protein acidic and rich in cysteine nor fibronectin. RNase protection assay demonstrated that mechanical strain induced all three alternatively spliced isoforms of tenascin-C. An angiotensin II receptor type 1 antagonist inhibited mechanical induction of brain natriuretic peptide but not tenascin-C. Antioxidants such as N-acetyl-L-cysteine, catalase, and 1,2-dihydroxy-benzene-3,5-disulfonate significantly inhibited induction of tenascin-C. Truncated tenascin-C promoter-reporter assays using dominant negative mutants of IB␣ and IB kinase ␤ and electrophoretic mobility shift assays indicated that mechanical strain increases tenascin-C gene transcription by activating nuclear factor-B through reactive oxygen species. Our findings demonstrate that mechanical strain induces tenascin-C in cardiac myocytes through a nuclear factor-B-dependent and angiotensin II-independent mechanism. These data also suggest that reactive oxygen species may participate in mechanically induced left ventricular remodeling.Cardiac hypertrophy is an independent risk factor of cardiac morbidity and mortality (1) and is characterized by an increase in myocyte mass and volume, as well as an increase of extracellular matrix proteins such as collagen (2). Angiotensin II is a potent stimulator of cardiac hypertrophy (3), and angiotensin-converting enzyme inhibitors prevent left ventricular hypertrophy in hypertensive animals and humans. For example, Kojima et al. (4) reported that treatment with TCV-116, an angiotensin II receptor type 1 (AT 1 ) 1 antagonist, decreased left ventricular weight, left ventricular wall thickness, and the transverse diameter of cardiac myocytes in spontaneously hypertensive rats.Recent studies (5, 6) indicate that angiotensin II-independent mechanisms may also mediate cardiac hypertrophy. Harada et al. (5) demonstrated that acute pressure overload could induce hypertrophic responses such as induction of c-fos, c-jun, and brain natriuretic peptide (BNP) gene expression, mitogen-activated protein (MAP) kinase activation, and increased heart weight/body weight, in the hearts of AT 1A knockout mice. Harada et al. (6) also reported that there were no significant differences between wild-type mice and AT 1A knockout mice in expression levels of fetal-type cardiac genes, in left ventricular wall thickness and systolic function, or in histological changes such as myocyte hypertrophy and fibrosis.

When cervical ectopic pregnancy is diagnosed early, US-guided termination or other conservative procedures allow preservation of the uterus, thus maintaining potential fertility.

No abstract

Urinary gonadotropin peptide (UGP; beta-core fragment), a major metabolite of human chorionic gonadotropin (hCG), was shown recently to be markedly elevated in Down syndrome pregnancy between 19 and 22 weeks of gestation. To confirm and extend this finding, we obtained maternal urine and matching maternal serum samples from 14 cases of Down syndrome and six other aneuploidies between 17 and 21 weeks of gestation. UGP was measured in all these samples and in 91 singleton control urines. Results were corrected for urinary creatinine level and expressed as multiples of the control median (MOM). hCG levels were assayed in all serum samples from the cases and compared with previously established reference values. The median UGP level in Down syndrome cases was 5.34 MOM (range 2.71-12.57); 88 per cent of the values were above the 95th centile of control levels after modelling. The median maternal serum hCG level for the same cases was 2.20 MOM (range 0.84-3.40); 36 per cent of the values were above the 95th centile. The level of UGP in every case including all other aneuploidies was higher than the comparable maternal serum hCG level. Elevated UGP measurements are strongly associated with fetal Down syndrome during the second trimester and could contribute to improved Down syndrome screening protocols that are more accessible and less expensive than are currently available.

Apoptosis of neurons and astrocytes is induced by human immunodeficiency type 1 (HIV-1) infection in vitro and has been demonstrated in brain tissue from patients with AIDS. We analyzed a panel of diverse HIV-1 primary isolates for the ability to replicate and induce neuronal and astrocyte apoptosis in primary human brain cultures. Apoptosis was induced three- to eightfold by infection with the blood-derived HIV-1 isolates 89.6, SG3, and ADA. In contrast, the brain-derived HIV-1 isolates YU2, JRFL, DS-br, RC-br, and KJ-br did not induce significant levels of apoptosis. The ability of HIV-1 isolates to induce apoptosis was independent of their replication capacity. Studies of recombinant chimeras between the SG3 and YU2 viruses showed that replacement of the YU2 Env with the SG3 Env was sufficient to confer the ability to induce apoptosis to the YU2 virus. Replacement of the Env V3 regions alone largely conferred the phenotypes of the parental clones. The SG3 Env used CXCR4 and CCR3 as coreceptors for virus entry, whereas YU2 used CCR5 and CCR3. The V3 regions of SG3 and YU2 conferred the ability to use CXCR4 and CCR5, respectively. In contrast, the 3′ region of Env, particularly the C3V4 region, was required in conjunction with the V3 region for efficient use of CCR3. These results provide evidence that Env is a major determinant of neurodegenerative mechanisms associated with HIV-1 infection in vitro and raise the possibility that blood-derived viruses which emerge during the late stages of disease may affect disease progression in the central nervous system.

We have successfully established mixed glial cell primary cultures prepared from individual fetal human brains (15-18 weeks' gestation in age). Cultures were maintained for as long as 3 months in either 10% fetal calf serum (FCS) or serum-free chemically defined medium (CDM). By morphological and immunohistochemical criteria, the precursor cell for human oligodendrocytes (O-2A cell) was identified. This cell exhibited the bipolar morphology and A2B5-positive (A2B5+) immunoreactivity typical of the O-2A precursor cell. With time in culture, cells possessing a stellate morphology appeared, some of which stained with the O4 antibody, indicative of cell differentiation in the oligodendroglial lineage. At yet older culture age, arborized cells bearing the O1 (galactocerebroside, GC) immunohistochemical marker and displaying the morphological characteristics typical of more mature oligodendrocytes were found, confirming their oligodendroglial identity. Oligodendroglial differentiation was supported best by CDM rather than FCS. To complement these observations, double immunofluorescent studies were performed on parietal sections from human fetal brains at 20 to 22 weeks of gestation. Bipolar A2B5+, multipolar A2B5+/O4+, and arborized A2B5-/O1+ cells were found, thus confirming the presence of oligodendrocytes in human fetal brain at this stage of prenatal development and consistent with the observations made in cell culture.

The thyroid of 31 fetuses at low risk for perinatal thyroid disease were evaluated sonographically. The transverse width and circumference of the fetal thyroid was measured prospectively to provide normative values for each gestational age. In addition, the thyroid of 23 fetuses at risk for thyroid disease were examined sonographically and compared to the control group. At birth, 18 of the neonates had no evidence of thyroid dysfunction, whereas 5 newborns had goiters and abnormal thyroid function. The fetal thyroid measurements for these 5 neonates were above the upper limit of the 95% confidence interval compared to the control group. The other 18 fetuses in the group at risk for thyroid disease but without evidence of thyroid dysfunction at birth had fetal thyroid measurement within the normal range.