Six corpora lutea (day 17 to day 25) of the menstrual cycle and four ovarian stromal tissues from seven cycling women were examined for the presence of epidermal growth factor (EGF) by ...immunocytochemistry with the use of the indirect peroxidase-antiperoxidase (PAP) procedure. After tissue sections of 4 μm were mounted, endogenous peroxidases were removed with hydrogen peroxide, and the sections were incubated for 1 hour at room temperature followed by 16 hours at 4°C with a highly specific antisera for mouse EGF (mEGF), nonimmunized normal rabbit serum, or antiserum preadsorbed with excess antigen. After the reaction with the second antibody (goat antirabbit IgG) for 1 hour at room temperature, the peptide was localized by use of PAP and 3.3′ -diaminobenzidine as the chromogen. EGF could be localized in the luteal cells of five corpora lutea (day 17 to day 25) but not in a corpus luteum of day 22 and all ovarian stromal tissue examined. The localization of immunoreactive EGF in the human corpus luteum demonstrates directly for the first time the presence of this growth-promoting peptide in this tissue, which suggests its local production.
We have demonstrated the presence of epidermal growth factor (EGF) and its receptors in human non-gestational corpora lutea. To determine further the characteristics of EGF receptor binding, we ...examined 30 human corpora lutea throughout the luteal phase and during pregnancy. Scatchard plots of EGF binding in 29 of the 30 corpora lutea were curvilinear, suggesting negative co-operativity. The mean +/- SE of the association constant Ka was (0.9 +/- 0.2) x 10(9) l/mol, the dissociation constant Kd was (2.2 +/- 0.3) x 10(-9) mol/l and the number of binding sites (Rt) was (15.8 +/- 2.1) x 10(-19) mol/micrograms protein for non-gestational corpora lutea. The Kd increased significantly in late pregnancies compared to early pregnancies (P = < 0.005), while Rt was significantly higher in term pregnancies than in either early pregnancy (P < 0.01) or the menstrual cycle (P < 0.001). Corpora lutea atretica (n = 2) and ovarian stroma (n = 6) did not show any EGF binding activity. Our findings demonstrate the presence of specific EGF receptors in human corpora lutea of both the menstrual cycle and pregnancy. The changes in EGF binding parameters in early pregnancy suggest that there may be a relationship between the role of EGF and ovarian steroidogenesis.
A multicenter study was conducted with the objective to evaluate a reverse line blot (RLB) assay to detect resistance to rifampin (RIF), isoniazid (INH), streptomycin (STR), and ethambutol (EMB) in ...clinical isolates of
Mycobacterium tuberculosis. Oligonucleotides specific for wild type and mutant (drug resistance linked) alleles of the selected codons in the genes
rpoB, inhA, ahpC, rpsL, rrs, embB, were immobilized on a nylon membrane. The RLB assay conditions were optimized following analysis of DNA samples with known sequences of the targeted genes. For validation of the method at different geographical locations, the membranes were sent to seven laboratories in six countries representing the regions with high burdens of multudrug-resistant tuberculosis. The reproducibility of the assay for detection of
rpoB genotypes was initially evaluated on a blinded set of twenty reference DNA samples with known allele types and overall concordant results were obtained. Further mutation analysis was performed by each laboratory on the local strains. Upon RLB analysis of 315 clinical isolates from different countries, 132 (85.2%) of 155 RIF-resistant and 28 (51.0%) of 55 EMB-resistant isolates were correctly identified, showing applicability of the assay when targeting the
rpoB hot-spot region and
embB306. Mutations in the
inhA and
ahpC promoter regions, conferring resistance to INH, were successfully identified in respectively 16.9% and 13.2% of INH-resistant strains. Likewise, mutations in
rrs513 and
rpsL88 that confer resistance to STR were identified in respectively 15.1% and 10.7% of STR-resistant strains. It should be mentioned that mutation analysis of the above targets usually requires rather costly DNA sequencing to which the proposed RLB assay presents rapid and inexpensive alternative. Furthermore, the proposed method requires the same simple equipment as that used for spoligotyping and permits simultaneous analysis of up to 40 samples. This technique is a first attempt to combine different targets in a single assay for prediction of antituberculosis drugs resistance. It is open to further development as it allows easy incorporation of new probes for detection of mutations in other genes associated with resistance to second-line (e.g., fluoroquinolones) and new antituberculosis compounds.
To determine whether the human corpus luteum is a source of relaxin and oxytocin, we measured the concentrations of these peptides in plasma obtained from the ovarian veins of ovaries with and ...without a corpus luteum and compared these to peripheral plasma levels. Peripheral and ovarian venous blood samples were obtained from 34 nonpregnant women, 13 during the luteal phase and 21 during the follicular phase of their cycles, and from a 6-week pregnant woman. Plasma relaxin, oxytocin, and progesterone concentrations were determined by sensitive and specific RIAs. Plasma relaxin levels were not detectable (less than 0.16 microgram/L) in peripheral or ovarian venous plasma not draining a corpus luteum. The mean relaxin concentration in plasma draining an ovary with a corpus luteum was 0.41 +/- 0.09 (+/- SE) microgram/L. Oxytocin levels also were significantly higher in plasma draining an ovary with a corpus luteum (6.70 +/- 1.86 pmol/L) than in that draining the ovary with no corpus luteum (1.58 +/- 0.09 pmol/L; P less than 0.01) or in peripheral plasma (1.58 +/- 0.09 pmol/L; P less than 0.025). The mean progesterone concentration also was highest in plasma draining an ovary with a corpus luteum (210.2 +/- 50.5 nmol/L) compared with those in plasma from the contralateral ovarian vein (40.3 +/- 16.5 nmol/L P less than 0.005) and peripheral plasma (30.2 +/- 5.7 nmol/L; P less than 0.005) during the luteal phase. In a woman who was 6 weeks pregnant, plasma draining the ovary with a corpus luteum had 1.9 micrograms relaxin/L, but only 0.49 pmol/L oxytocin; the latter was similar to concentrations in noncorpus luteum-bearing ovarian venous plasma. These findings indicate that the human corpus luteum secretes relaxin, oxytocin, and progesterone. Both ovarian oxytocin and relaxin may function as paracrine or autocrine modulators of luteal function.
We report on a high-statistics measurement of the most basic double-pionic fusion reaction over the energy region of the d (*)(2380) resonance by use of a polarized deuteron beam and observing the ...double fusion reaction in the quasifree scattering mode. The measurements were performed with the WASA detector setup at COSY. The data reveal substantial analyzing powers and confirm conclusions about the d(*) resonance obtained from unpolarized measurements. We also confirm the previous unpolarized data obtained under complementary kinematic conditions.
Selected, peer reviewed papers from the International Conference on Applications and Design in Mechanical Engineering (ICADME 2015), March 30-31, 2015, Kuala Lumpur, Malaysia.
The radiation-driven, low-adiabat, cryogenic DT layered plastic capsule implosions were carried out on the National Ignition Facility (NIF) to study the sensitivity of performance to peak power and ...drive duration. An implosion with extended drive and at reduced peak power of 350 TW achieved the highest compression with fuel areal density of ~1.3±0.1 g/cm 2, representing a significant step from previously measured ~1.0 g/cm2 toward a goal of 1.5 g/cm 2. Moreover, for future experiments will focus on understanding and mitigating hydrodynamic instabilities and mix, and improving symmetry required to reach the threshold for thermonuclear ignition on NIF.
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