bluuebunny/crossref_metadata_2025_split huggingface dataset:
Optimization of Nisin Production by Lactococcus lactis UQ2 Using Supplemented Whey as Alternative Culture Medium
Authors: González‐Toledo S.Y. , Domínguez‐Domínguez J. , García‐Almendárez B.E. , Prado‐Barragán L.A. , Regalado‐González C. 
Published: July 2010

Abstract:  Lactococcus lactis UQ2 is a nisin A‐producing native strain. In the present study, the production of nisin by L. lactis UQ2 in a bioreactor using supplemented sweet whey (SW) was optimized by a statistical design of experiments and response surface methodology (RSM). In a 1st approach, a fractional factorial design (FFD) of the order 25‐1 with 3 central points was used. The effect on nisin production of air flow, SW, soybean peptone (SP), MgSO4/MnSO4 mixture, and Tween 80 was evaluated. From FFD, the most significant factors affecting nisin production were SP (P = 0.011), and SW (P = 0.037). To find optimum conditions, a central composite design (CCD) with 2 central points was used. Three factors were considered, SW (7 to 10 g/L), SP (7 to10 g/L), and small amounts of added nisin as self‐inducer (NI 34.4 to 74.4 IU/L). Nisin production was expressed as international units (IU). From RSM, an optimum nisin activity of 180 IU/mL was predicted at 74.4 IU/L NI, 13.8 g/L SP, and 14.9 or 5.11 g/L SW, while confirmatory experiments showed a maximum activity of 178 ± 5.2 IU/mL, verifying the validity of the model. The 2nd‐order model showed a coefficient of determination (R2) of 0.828. Optimized conditions were used for constant pH fermentations, where a maximum activity of 575 ± 17 IU/mL was achieved at pH 6.5 after 12 h. The adsorption‐desorption technique was used to partially purify nisin, followed by drying. The resulting powder showed an activity of 102150 IU/g. Practical Application: Nisin production was optimized using supplemented whey as alternative culture medium, using a native L. lactis UQ2 strain. Soybean peptone, SW, and subinhibitory amounts of nisin were successfully employed to optimize nisin production by L. lactis UQ2. Dried semipurified nisin showed an activity of 102150 IU/g.
Effect of NO synthase inhibition on retinal vessel reaction to isometric exercise in healthy humans
Authors: Lasta Michael , Polak Kaija , Luksch Alexandra , Garhofer Gerhard , Schmetterer Leopold 
Published: July 2010

Abstract.Purpose:  It has been shown that retinal blood flow is autoregulated, meaning that flow is independent of perfusion pressure within a certain range. We tested the hypothesis that nitric oxide (NO) synthase inhibition alters the response of retinal arterial and venous vessels during isometric exercise.Methods:  In this study, nine healthy subjects were included. Each subject received the NO synthase inhibitor Ng‐monomethyl‐l‐Arginine (l‐NMMA, the α‐receptor agonist phenylephrine or placebo intravenously on three study days. Retinal vessel diameter was assessed with the retinal vessel analyser (RVA), at baseline and during a squatting period of 6–7 min in absence or presence of l‐NMMA, phenylephrine or placebo.Results:  Mean arterial pressure (MAP) and pulse rate (PR) increased significantly during all pretreatment squatting periods (p < 0.001) Retinal venous and arterial diameters showed a continuous decrease during squatting (p < 0.001). Phenylephrine increased MAP and PR but did not alter the retinal vessel diameter response to squatting. Administration of l‐NMMA lead to a significant decrease in venous diameter before isometric exercise (p = 0.004). In addition, the retinal venous diameter response during administration of the NO synthase inhibitor was less pronounced than during phenylephrine or placebo (p < 0.001).Conclusion:  Our study confirms that NO plays an important role in the control of retinal vascular tone at rest. In addition, the present data indicate a role of NO in retinal autoregulation, because the response of retinal venous diameters was altered after NO synthase inhibition. The nature of involvement, however, appears to be complex and requires further studies.
Microneedle Delivery of H5N1 Influenza Virus-Like Particles to the Skin Induces Long-Lasting B- and T-Cell Responses in Mice
Authors: Song Jae-Min , Kim Yeu-Chun , Lipatov Aleksandr S. , Pearton Marc , Davis C. Todd , Yoo Dae-Goon , Park Kyoung-Mi , Chen Li-Mei , Quan Fu-Shi , Birchall James C. , Donis Ruben O. , Prausnitz Mark R. , Compans Richard W. , Kang Sang-Moo 
Published: July 2010

ABSTRACTA simple method suitable for self-administration of vaccine would improve mass immunization, particularly during a pandemic outbreak. Influenza virus-like particles (VLPs) have been suggested as promising vaccine candidates against potentially pandemic influenza viruses, as they confer long-lasting immunity but are not infectious. We investigated the immunogenicity and protective efficacy of influenza H5 VLPs containing the hemagglutinin (HA) of A/Vietnam/1203/04 (H5N1) virus delivered into the skin of mice using metal microneedle patches and also studied the response of Langerhans cells in a human skin model. Prime-boost microneedle vaccinations with H5 VLPs elicited higher levels of virus-specific IgG1 and IgG2a antibodies, virus-specific antibody-secreting cells, and cytokine-producing cells up to 8 months after vaccination compared to the same antigen delivered intramuscularly. Both prime-boost microneedle and intramuscular vaccinations with H5 VLPs induced similar hemagglutination inhibition titers and conferred 100% protection against lethal challenge with the wild-type A/Vietnam/1203/04 virus 16 weeks after vaccination. Microneedle delivery of influenza VLPs to viable human skin using microneedles induced the movement of CD207+Langerhans cells toward the basement membrane. Microneedle vaccination in the skin with H5 VLPs represents a promising approach for a self-administered vaccine against viruses with pandemic potential.
Development and Application of Multiprobe Real-Time PCR Method Targeting the
hsp65
Gene for Differentiation of
Mycobacterium
Species from Isolates and Sputum Specimens
Authors: Kim Kijeong , Lee Hyungki , Lee Mi-Kyung , Lee Seoung-Ae , Shim Tae-Sun , Lim Seong Yong , Koh Won-Jung , Yim Jae-Joon , Munkhtsetseg Bazarragchaa , Kim Wonyong , Chung Sang-In , Kook Yoon-Hoh , Kim Bum-Joon 
Published: July 2010

ABSTRACT

We developed a multiprobe real-time PCR assay targeting
hsp65
(HMPRT-PCR) to detect and identify mycobacterial isolates and isolates directly from sputum specimens. Primers and probes for HMPRT-PCR were designed on the basis of the
hsp65
gene sequence, enabling the recognition of seven pathogenic mycobacteria, including
Mycobacterium tuberculosis
,
M. avium
,
M. intracellulare
,
M. kansasii
,
M. abscessus
,
M. massiliense
, and
M. fortuitum.
This technique was applied to 24 reference and 133 clinical isolates and differentiated between all strains with 100% sensitivity and specificity. Furthermore, this method was applied to sputum specimens from 117 consecutive smear-positive patients with smear results of from a trace to 3+. These results were then compared to those obtained using the
rpoB
PCR-restriction analysis method with samples from cultures of the same sputum specimens. The HMPRT-PCR method correctly identified the mycobacteria in 89 samples (76.0%, 89/117), and moreover, the sensitivity level was increased to 94.3% (50/53) for sputa with an acid-fast bacillus score equal to or greater than 2+. Our data suggest that this novel HMPRT-PCR method could be a promising approach for detecting pathogenic mycobacterial species from sputum samples and culture isolates routinely in a clinical setting.
TMPRSS2 and TMPRSS4 Facilitate Trypsin-Independent Spread of Influenza Virus in Caco-2 Cells
Authors: Bertram Stephanie , Glowacka Ilona , Blazejewska Paulina , Soilleux Elizabeth , Allen Paul , Danisch Simon , Steffen Imke , Choi So-Young , Park Youngwoo , Schneider Heike , Schughart Klaus , Pöhlmann Stefan 
Published: July 2010

ABSTRACT
Proteolysis of influenza virus hemagglutinin by host cell proteases is essential for viral infectivity, but the proteases responsible are not well defined. Recently, we showed that engineered expression of the type II transmembrane serine proteases (TTSPs) TMPRSS2 and TMPRSS4 allows hemagglutinin (HA) cleavage. Here we analyzed whether TMPRSS2 and TMPRSS4 are expressed in influenza virus target cells and support viral spread in the absence of exogenously added protease (trypsin). We found that transient expression of TMPRSS2 and TMPRSS4 resulted in HA cleavage and trypsin-independent viral spread. Endogenous expression of TMPRSS2 and TMPRSS4 in cell lines correlated with the ability to support the spread of influenza virus in the absence of trypsin, indicating that these proteases might activate influenza virus in naturally permissive cells. Indeed, RNA interference (RNAi)-mediated knockdown of both TMPRSS2 and TMPRSS4 in Caco-2 cells, which released fully infectious virus without trypsin treatment, markedly reduced the spread of influenza virus, demonstrating that these proteases were responsible for efficient proteolytic activation of HA in this cell line. Finally, TMPRSS2 was found to be coexpressed with the major receptor determinant of human influenza viruses, 2,6-linked sialic acids, in human alveolar epithelium, indicating that viral target cells in the human respiratory tract express TMPRSS2. Collectively, our results point toward an important role for TMPRSS2 and possibly TMPRSS4 in influenza virus replication and highlight the former protease as a potential therapeutic target.
Yellow Fever Virus Maintenance in Trinidad and Its Dispersal throughout the Americas
Authors: Auguste Albert J. , Lemey Philippe , Pybus Oliver G. , Suchard Marc A. , Salas Rosa Alba , Adesiyun Abiodun A. , Barrett Alan D. , Tesh Robert B. , Weaver Scott C. , Carrington Christine V. F. 
Published: July 2010

ABSTRACT

Trinidad, like many other American regions, experiences repeated epizootics of yellow fever virus (YFV). However, it is unclear whether these result from
in situ
evolution (enzootic maintenance) or regular reintroduction of YFV from the South American mainland. To discriminate between these hypotheses, we carried out a Bayesian phylogeographic analysis of over 100 prM/E gene sequences sampled from 8 South American countries. These included newly sequenced isolates from the recent 2008-2009 Trinidad epizootic and isolates derived from mainland countries within the last decade. The results indicate that the most recent common ancestor of the 2008-2009 epizootic existed in Trinidad 4.2 years prior to 2009 (95% highest probability density [HPD], 0.5 to 9.0 years). Our data also suggest a Trinidad origin for the progenitor of the 1995 Trinidad epizootic and support
in situ
evolution of YFV between the 1979 and 1988-1989 Trinidad epizootics. Using the same phylogeographic approach, we also inferred the historical spread of YFV in the Americas. The results suggest a Brazilian origin for YFV in the Americas and an overall dispersal rate of 182 km/year (95% HPD, 52 to 462 km/year), with Brazil as the major source population for surrounding countries. There is also strong statistical support for epidemiological links between four Brazilian regions and other countries. In contrast, while there were well-supported epidemiological links within Peru, the only statistically supported external link was a relatively weak link with neighboring Bolivia. Lastly, we performed a complete analysis of the genome of a newly sequenced Trinidad 2009 isolate, the first complete genome for a genotype I YFV isolate.
Interaction of the Papillomavirus E8∧E2C Protein with the Cellular CHD6 Protein Contributes to Transcriptional Repression
Authors: Fertey Jasmin , Ammermann Ingo , Winkler Michael , Stöger Reinhard , Iftner Thomas , Stubenrauch Frank 
Published: July 2010

ABSTRACT
Expression of the E6 and E7 oncogenes of high-risk human papillomaviruses (HPV) is controlled by cellular transcription factors and by viral E2 and E8∧E2C proteins, which are both derived from the HPV E2 gene. Both proteins bind to and repress the HPV E6/E7 promoter. Promoter inhibition has been suggested to be due to binding site competition with cellular transcription factors and to interactions of different cellular transcription modulators with the different amino termini of E2 and E8∧E2C. We have now identified the cellular chromodomain helicase DNA binding domain 6 protein (CHD6) as a novel interactor with HPV31 E8∧E2C by using yeast two-hybrid screening. Pull-down and coimmunoprecipitation assays indicate that CHD6 interacts with the HPV31 E8∧E2C protein via the E2C domain. This interaction is conserved, as it occurs also with the E8∧E2C proteins expressed by HPV16 and -18 and with the HPV31 E2 protein. Both RNA knockdown experiments and mutational analyses of the E2C domain suggest that binding of CHD6 to E8∧E2C contributes to the transcriptional repression of the HPV E6/E7 oncogene promoter. We provide evidence that CHD6 is also involved in transcriptional repression but not activation by E2. Taken together our results indicate that the E2C domain not only mediates specific DNA binding but also has an additional role in transcriptional repression by recruitment of the CHD6 protein. This suggests that repression of the E6/E7 promoter by E2 and E8∧E2C involves multiple interactions with host cell proteins through different protein domains.
The Human Papillomavirus Type 16 E6 Oncoprotein Activates mTORC1 Signaling and Increases Protein Synthesis
Authors: Spangle Jennifer M. , Münger Karl 
Published: July 2010

ABSTRACT
The mammalian target of rapamycin (mTOR) kinase acts as a cellular rheostat that integrates signals from a variety of cellular signal transduction pathways that sense growth factor and nutrient availability as well as intracellular energy status. It was previously reported that the human papillomavirus type 16 (HPV16) E6 oncoprotein may activate the S6 protein kinase (S6K) through binding and E6AP-mediated degradation of the mTOR inhibitor tuberous sclerosis complex 2 (TSC2) (Z. Lu, X. Hu, Y. Li, L. Zheng, Y. Zhou, H. Jiang, T. Ning, Z. Basang, C. Zhang, and Y. Ke, J. Biol. Chem. 279:35664-35670, 2004; L. Zheng, H. Ding, Z. Lu, Y. Li, Y. Pan, T. Ning, and Y. Ke, Genes Cells 13:285-294, 2008). Our results confirmed that HPV16 E6 expression causes an increase in mTORC1 activity through enhanced phosphorylation of mTOR and activation of downstream signaling pathways S6K and eukaryotic initiation factor binding protein 1 (4E-BP1). However, we did not detect a decrease in TSC2 levels in HPV16 E6-expressing cells. We discovered, however, that HPV16 E6 expression causes AKT activation through the upstream kinases PDK1 and mTORC2 under conditions of nutrient deprivation. We show that HPV16 E6 expression causes an increase in protein synthesis by enhancing translation initiation complex assembly at the 5′ mRNA cap and an increase in cap-dependent translation. The increase in cap-dependent translation likely results from HPV16 E6-induced AKT/mTORC1 activation, as the assembly of the translation initiation complex and cap-dependent translation are rapamycin sensitive. Lastly, coexpression of the HPV16 E6 and E7 oncoproteins does not affect HPV16 E6-induced activation of mTORC1 and cap-dependent translation. HPV16 E6-mediated activation of mTORC1 signaling and cap-dependent translation may be a mechanism to promote viral replication under conditions of limited nutrient supply in differentiated, HPV oncoprotein-expressing proliferating cells.
Role of Herpes Simplex Virus ICP27 in the Degradation of mRNA by Virion Host Shutoff RNase
Authors: Taddeo Brunella , Zhang Weiran , Roizman Bernard 
Published: July 2010

ABSTRACT
The virion host shutoff (VHS) RNase tegument protein released into cells by infecting virus has two effects. Preexisting stable mRNAs (e.g., GAPDH [glyceraldehyde-3-phosphate dehydrogenase]) are rapidly degraded. Stress response RNAs containing AU-rich elements (AREs) in the 3′ untranslated region (3′UTR) are deadenylated and cleaved, but the cleavage products persist for hours, in contrast to the short half-lives of ARE-containing mRNAs in uninfected cells. At late times, the VHS RNase is neutralized by the viral structural proteins VP16 and VP22. A recent study (J. A. Corcoran, W. L. Hsu, and J. R. Smiley, J. Virol. 80:9720-9729, 2006) reported that, at relatively late times after infection, ARE RNAs are rapidly degraded in cells infected with ΔICP27 mutant virus and concluded that ICP27 “stabilizes” ARE mRNAs. We report the following. (i) The rates of degradation of ARE mRNA at early times (3 h) after infection with the wild type or the ΔICP27 mutant virus are virtually identical, and hence ICP27 plays no role in this process. (ii) In noncomplementing cells, VHS RNase or VP22 is not synthesized. Therefore, the only VHS that is active is brought into cells by the ΔICP27 mutant. (ii) The VHS RNase brought into the cells by the ΔICP27 virus is reduced in potency relative to that of wild-type virus. Hence the rapid degradation of ARE mRNAs noted in ΔICP27 mutant-infected cells at late times is similar to that taking place in mock-infected or in ΔVHS RNase mutant-virus-infected cells and does not by itself support the hypothesis that ICP27 stabilizes ARE mRNAs. (iii) Concurrently, we present the first evidence that VHS RNase interacts with ICP27 most likely when bound to cap- and poly(A)-binding proteins, respectively.
Chronic kidney disease presenting acutely: presentation, clinical features and outcome of patients with irreversible chronic kidney disease who require dialysis immediately
Authors: Wolfe Maytal , Almond Alison , Robertson Sue , Donaldson Ken , Isles Chris 
Published: July 2010

Abstract

Background
Patients with irreversible chronic kidney disease who require dialysis immediately are a subset of ultra late referrals for whom the term chronic kidney disease presenting acutely might usefully be applied. Although well known to nephrologists and recognised as a specific group with considerable problems, little has been written about them.

Objective
To describe the presentation, clinical features and outcome of irreversible chronic kidney disease presenting acutely, with particular reference to nausea and vomiting as presenting symptoms.

Method and results
Review of 202 consecutive patients with irreversible chronic kidney disease who had dialysis between 1996 and 2006 showed that 15 (7%) had required dialysis immediately or within 7 days of presentation. Analysis of 14 available case records showed eight avoidable late referrals: previous evidence of renal failure in six, and two patients with diabetes who had not had their renal function checked. Gastrointestinal symptoms were common and led to further delays in diagnosis, with three patients having endoscopy requested before their bloods were checked. Physical and psychological morbidity associated with this form of presentation was high.

Conclusions
Chronic kidney disease presenting acutely is not uncommon, often avoidable and associated with adverse outcomes. The identification, follow-up and appropriate referral of patients with raised serum creatinine is likely to reduce its incidence. Nausea and vomiting may occur sufficiently frequently in advanced renal failure to justify measuring renal function before proceeding to endoscopy when patients present with gastrointestinal symptoms.
Time trends of female breast cancer mortality in Greece during 1980–2005: a population based study
Authors: Pavlidou E , Zafrakas M , Papadakis N , Agorastos T , Benos A 
Published: July 2010

Abstract

Background
Changes in female breast cancer mortality trends have been observed in recent years in western countries. The aim of the present study was to analyse breast cancer mortality in Greece, between 1980 and 2005.

Methods
Time trends of breast cancer mortality were calculated per 100 000 women in the whole female population of Greece, in different age groups, and in different areas of the country. Mortality data and population age distribution were provided by the National Statistical Service of Greece.

Results
Overall, female breast cancer mortality in Greece had an increasing trend during 1980–2005. Subgroup analysis according to age showed that this increase was confined to women older than 70 years. In contrast, a mild decrease was noted after the mid-1990s in women 40–69 years old. There were no notable regional differences in breast cancer mortality.

Conclusions
The increasing trend of breast cancer mortality in women aged 70 years and older could be attributed to limited use of secondary prevention methods and rare administration of systemic cytotoxic chemotherapy in these women. In contrast, implementation of these strategies could explain the recent reduction of breast cancer mortality in younger age groups.
Republished paper: Determinants of child health and development: the contribution of ALSPAC—a personal view of the birth cohort study
Authors: Golding Jean 
Published: July 2010

Abstract
Britain has a unique experience of national longitudinal birth cohorts, but the Avon Longitudinal Study of Parents and Children differed in two major respects—it was based in one area rather than being a national sample, and it started in pregnancy rather than at birth or later in the first year. This paper outlines a personal selection of 10 topics, highlighting results from some of the 400+ papers that have already been published from this study. It indicates in particular how many childcare and domestic fashions were neither of benefit to the children (or their parents), the importance of pregnancy in regard to childhood conditions and the likely dangers of some common chemicals, whether in medications or domestic products.
Secure and robust virtual coordinate system in wireless sensor networks
Authors: Dong Jing , Ackermann Kurt E. , Bavar Brett , Nita-Rotaru Cristina 
Published: July 2010

Virtual Coordinate System (VCS)-based routing provides a practical, efficient, and scalable means for point-to-point routing in wireless sensor networks. Several VCS-based routing protocols have been proposed in the last few years, all assuming that nodes behave correctly. However, many applications require deploying sensor networks in adversarial environments, making VCS-based routing protocols vulnerable to numerous attacks.
In this article, we study the security of VCS-based routing protocols, with a focus on the unique component of VCS-based routing protocols, the virtual coordinate system. We first identify the security requirements of a correctly functioning VCS-based routing protocol and a set of novel attacks that can result in the violation of each of the identified requirements. The attacks target the underlying virtual coordinate system and can be mounted with low resources. However, they are epidemic in nature and are highly destructive to system performance. We then propose lightweight defense mechanisms designed specifically for resource-constrained sensor networks against each of the identified attacks. The proposed techniques require only local information on sensor nodes and take into account the unreliable nature of wireless links and network churn. Finally, we evaluate experimentally the impact of the attacks and the effectiveness of our defense mechanisms using a well-known VCS-based routing protocol, BVR. Our experiments show that the proposed techniques successfully mitigate all the identified attacks under a realistic link model and even at a high level of network churn.