Middle East respiratory syndrome coronavirus not detected in children hospitalized with acute respiratory illness in Amman, Jordan, March 2010 to September 2012.

Hospitalized children < 2 years of age in Amman, Jordan, admitted for fever and/or respiratory symptoms, were tested for Middle East respiratory syndrome coronavirus (MERS-CoV): MERS-CoV by real-time RT-PCR (rRT-PCR). This was a prospective year-round viral surveillance study in children <2 years of age admitted with acute respiratory symptoms and/or fever from March 2010 to September 2012 and enrolled from a government-run hospital, Al-Bashir in Amman, Jordan. Clinical and demographic data, including antibiotic use, were collected. Combined nasal/throat swabs were collected, aliquoted, and frozen at -80°C. Specimen aliquots were shipped to Vanderbilt University and the Centers for Disease Control and Prevention (CDC), and tested by rRT-PCR for MERS-CoV. Of the 2433 subjects enrolled from 16 March 2010 to 10 September 2012, 2427 subjects had viral testing and clinical data. Of 1898 specimens prospectively tested for other viruses between 16 March 2010 and 18 March 2012, 474 samples did not have other common respiratory viruses detected. These samples were tested at CDC for MERS-CoV and all were negative by rRT-PCR for MERS-CoV. Of the remaining 531 samples, collected from 19 March 2012 to 10 September 2012 and tested at Vanderbilt, none were positive for MERS-CoV. Our negative findings from a large sample of young Jordanian children hospitalized with fever and/or respiratory symptoms suggest that MERS-CoV was not widely circulating in Amman, Jordan, during the 30-month period of prospective, active surveillance occurring before and after the first documented MERS-CoV outbreak in the Middle East region.

Inhibition of transforming growth factor beta signaling in MCF-7 cells results in resistance to tumor necrosis factor alpha: a role for Bcl-2.

Transforming growth factor beta (TGF-beta) is a multifunctional cytokine capable of regulating diverse cellular processes. In this study we investigated the effect of autocrine TGF-beta signaling on tumor necrosis factor (TNF) alpha-induced cell death. We abrogated the TGF-beta autocrine loop by overexpression of a truncated TGF-beta type II receptor in MCF-7 breast carcinoma cells and found that this generated resistance to TNF-alpha-induced cytotoxicity. To elucidate the molecular basis of the influence of TGF-beta on TNF-alpha-induced cytotoxicity, we evaluated the expression levels or activities of proteins involved in TNF-alpha signal transduction or the regulation of apoptosis in general in TGF-beta-responsive and TGF-beta-nonresponsive MCF-7 cells. We observed no significant difference in the expression of TNF-alpha receptors or the TNF receptor-associated death domain protein. In addition, downstream activation of nuclear factor kappaB by TNF-alpha was not altered in cells that had lost TGF-beta responsiveness. Analysis of members of the Bcl-2 family of apoptosis-regulatory proteins revealed that Bcl-X(L) and Bax expression levels were not changed by disruption of TGF-beta signaling. In contrast, the TGF-beta-nonresponsive cells expressed much higher levels of Bcl-2 protein and mRNA than did cells with an intact TGF-beta autocrine loop. Furthermore, restoration of a TGF-beta signal to MCF-7 cells that had spontaneously acquired resistance to TGF-beta caused a reduction in Bcl-2 protein expression. Taken together, our data indicate that loss of autocrine TGF-beta signaling results in enhanced resistance to TNF-alpha-mediated cell death and that this is likely to be mediated by derepression of Bcl-2 expression.

Stress and nursing in the pig: role of HPA axis and endogenous opioid peptides.

To determine the effect of stress on nursing, and the roles of HPA activity and opioid peptides, nine sows had their piglets removed for 2 h and were treated as follows: (a) control; (b) nose-snare restraint for 20 min; (c) naloxone injections (i.v. 2 mg/kg); and (d) snare + naloxone. After the treatment, the piglets were returned, milk ejections were timed, and the sows' blood sampled every 10 min for cortisol, growth hormone (GH), and prolactin assays. Piglet removal increased cortisol and decreased prolactin and GH. This was reversed when the piglets were returned. Restraint increased cortisol and decreased GH, but did not affect prolactin. Naloxone alone increased cortisol and decreased GH but did not increase the effect of restraint. The rise in GH following the piglets' return was abolished by the combination of restraint and naloxone. Neither restraint nor naloxone delayed the latency to first milk ejection or reduced the frequency. No unsuccessful nursings were observed. First milk ejections occurred when cortisol levels were elevated. Stress-induced activity of the hypothalamo-pituitary-adrenocortical (HPA) axis does not inhibit milk ejection in the pig, but this is not due to a protective opioid action. Endogenous opioids protect lactogenic hormones against inhibition by stress.

Acidic thiol proteinase activity of Schistosoma mansoni cultured in vitro.

Schistosoma mansoni cultured in vitro for periods from 12 days to 33 weeks were analyzed for proteinase activity using a fluorescent substrate and compared with adult S. mansoni worms from infected mice. The enzyme activity measured was that of an acidic thiol-dependent proteinase previously described. Proteinase activity appears early in development, and rises rapidly to an approximately constant proportion of the total extractable worm protein. Cultured worms have 5 to 20 times higher specific activity than in vivo-developed worms. However, cultured worms and in vivo worms have approximately equivalent activities per worm.

Correlation between tumour size, metastatic spread and galactosyl transferase activity in cyclophosphamide-treated mice bearing the Lewis lung carcinoma.

Galactosyl transferase activity was measured in tumour and normal tissues of mice receiving cyclophosphamide treatment for Lewis lung carcinoma. Animals which responded to cyclophosphamide therapy had significantly smaller tumours with fewer metastases than the untreated mice. The level of galactosyl transferase was significantly reduced in the tumours which were inhibited by the cyclophosphamide treatment.