bluuebunny/crossref_metadata_2025 huggingface dataset:
Short‐chain chitin oligomers from arbuscular mycorrhizal fungi trigger nuclear Ca2+ spiking in Medicago truncatula roots and their production is enhanced by strigolactone
Authors: Genre Andrea , Chabaud Mireille , Balzergue Coline , Puech‐Pagès Virginie , Novero Mara , Rey Thomas , Fournier Joëlle , Rochange Soizic , Bécard Guillaume , Bonfante Paola , Barker David G.
Published: February 2013

The primary objective of this study was to identify the molecular signals present in arbuscular mycorrhizal (AM) germinated spore exudates (GSEs) responsible for activating nuclear Ca2+ spiking in the Medicago truncatula root epidermis.

Medicago truncatula root organ cultures (ROCs) expressing a nuclear‐localized cameleon reporter were used as a bioassay to detect AM‐associated Ca2+ spiking responses and LC‐MS to characterize targeted molecules in GSEs.

This approach has revealed that short‐chain chitin oligomers (COs) can mimic AM GSE‐elicited Ca2+ spiking, with maximum activity observed for CO4 and CO5. This spiking response is dependent on genes of the common SYM signalling pathway (DMI1/DMI2) but not on NFP, the putative Sinorhizobium meliloti Nod factor receptor. A major increase in the CO4/5 concentration in fungal exudates is observed when Rhizophagus irregularis spores are germinated in the presence of the synthetic strigolactone analogue GR24. By comparison with COs, both sulphated and nonsulphated Myc lipochito‐oligosaccharides (LCOs) are less efficient elicitors of Ca2+ spiking in M. truncatula ROCs.

We propose that short‐chain COs secreted by AM fungi are part of a molecular exchange with the host plant and that their perception in the epidermis leads to the activation of a SYM‐dependent signalling pathway involved in the initial stages of fungal root colonization.
Experiences, understandings and support needs of family carers of older patients with delirium: a descriptive mixed methods study in a hospital delirium unit
Authors: Toye Christine , Matthews Anne , Hill Andrew , Maher Sean
Published: February 2013

Background

Delirium is common in older patients. Little is known of support needs of families of older hospital patients with delirium.

Aims and objectives

To inform nursing practice, we sought to describe families' experiences, understanding of delirium and delirium care, and support needs.

Design

Descriptive mixed methods.

Methods

Review of questionnaire items used with families of people with terminal delirium informed development of a new questionnaire to evaluate the support needs in our study population. In a tertiary hospital delirium unit, we recruited 17 family carers of older patients with (non‐terminal) delirium to respond to this questionnaire. Twelve participants (11 female) also took part in interviews addressing the study's aims. Descriptive statistics were calculated and thematic analysis was undertaken.

Results

From the survey, key family issues included distress about the patient's condition, worries about future care, and a need for more information about how the patient might feel and how families could support the patient. Themes from interviews included The admission experience, Worries and concerns, Feeling supported, and The discharge experience. Limited understanding of delirium underpinned all themes. Families experienced shock and sadness at the change in the patient; they were reassured by the specialist care but needed more information about delirium, its effects and outcomes, and how they could help with care. Meeting long‐term postdischarge needs was a key concern.

Conclusions

In this study, families with a hospitalised older relative who had delirium described a distressing experience and needs for informational support. Further research is needed outside of delirium‐specific units and in samples including a greater proportion of male relatives.

Implications for practice

Nurses should work within the interdisciplinary team to ensure a planned, sensitive and timely approach to informing the patient's family about delirium and its implications for their relative, recognising the family carer's role and likely distress.
Bell‐Polynomial Approach and Integrability for the Coupled Gross–Pitaevskii Equations in Bose–Einstein Condensates
Authors: Wang Yu‐Feng , Tian Bo , Wang Ming
Published: February 2013

Under investigation in this paper are the coupled Gross–Pitaevskii equations, which describe the dynamics of two‐component Bose–Einstein condensates. Infinitely many conservation laws are obtained based on the Lax pair. Via the Hirota method, Bell‐polynomial approach and symbolic computation, bilinear forms, Bell‐polynomial‐typed transformation, and bilinear‐typed Bäcklund transformation are also derived. One‐ and two‐soliton‐like solutions are expressed explicitly. The gain/loss coefficient G(t) can influence the velocity of the solitonic envelopes. Head‐on and overtaking elastic interactions are shown and analyzed. Inelastic interactions between two soliton‐like envelopes are presented as well.
Bottled memories: On how alcohol affects eyewitness recall
Authors: Hagsand Angelica , Hjelmsäter Emma Roos Af , Granhag Pär Anders , Fahlke Claudia , Söderpalm‐Gordh Anna
Published: February 2013

This study investigated how different doses of alcohol affected eyewitness recall. Participants (N = 126) were randomly assigned to three groups with different blood alcohol concentration (BAC), either a control group (mean BAC 0.00%, N = 42), a lower alcohol dose group (mean BAC 0.04%, N = 40), or a higher alcohol dose group (mean BAC 0.06%, N = 44). After consumption, participants witnessed a movie of a mock crime and were interviewed one week later. The main results showed that witnesses with the higher intoxication level recalled fewer details compared to witnesses with the lower intoxication level. The amount of alcohol consumed did not have an impact on the accuracy rate. No sex differences were found. The results are discussed in the light of past research. We conclude that more studies are needed before recommendations can be made to an applied setting.
Long term cryopreservation in 5% DMSO maintains unchanged CD34+ cells viability and allows satisfactory hematological engraftment after peripheral blood stem cell transplantation
Authors: Abbruzzese L. , Agostini F. , Durante C. , Toffola R. T. , Rupolo M. , Rossi F. M. , Lleshi A. , Zanolin S. , Michieli M. , Mazzucato M.
Published: February 2013

Peripheral blood stem cell cryopreservation is associated with cell damage and decreased viability. We evaluated the impact of up to 10 years of cryopreservation (5% DMSO) on viability of CD34+ cells utilizing graft samples of consecutive patients (2002–2012) with different malignancies who underwent stem cell collection and transplantation. Viability of CD34+ cells from oncohaematological patients measured after 5 weeks (97·2 ± 0·6%) or after 9–10 years of cryopreservation (95·9 ± 0·5%) was unaffected. Haemoglobin, granulocyte and platelet recovery after transplantation of long‐term cryopreserved grafts occurred within 8–13 days. CD34+ stem cells can be safely stored up to 9–10 years, without affecting cell viability and clinical effectiveness.
Standardization of CD62P measurement: results of an international comparative study
Authors: Levin E. , Serrano K. , Devine D. V. ,   for the Biomedical Excellence for Safer Transfusion (BEST) Collaborative
Published: February 2013

Background and Objectives

Despite long being a mainstay in describing platelet activation via degranulation, interlaboratory variation remains an issue in measurement of membrane CD62P by flow cytometry. Our objective was to identify actions that may minimize this variation.

Materials and Methods

Sixteen laboratories participated in an international comparative study. Two sets of platelet samples were prepared in one laboratory. Set 1 was stained and fixed; set 2 was fixed and required staining at participating laboratories. A single‐staining method was used, and platelet populations were selected based on forward scatter/side scatter characteristics. Calibration beads were used to standardize measurement across different instruments.

Results

There was a large discrepancy in reported CD62P values among study sites [interlaboratory coefficient of variance (CV): 36–78%]. When electronic data were re‐analysed by a single analyst using a consistent gating strategy and a stable reference point, variation decreased markedly (CV < 12%), indicating a problem with isotype control samples, possibly related to sample fixation or shipment.

Conclusion

Consensus regarding gating strategies and use of a reliable reference point would greatly improve agreement in interlaboratory CD62P measurement.
Multi‐transgenic pigs for xenoislet transplantation
Authors: Ayares David , Phelps Carol , Vaught Todd , Ball Suyapa , Monahan Jeff , Walters Anneke , Giraldo Angelica , Bertera Suzanne , van der Windt Dirk , Wijkstrom Martin , Cooper David K. C. , Bottino Rita , Trucco Massimo
Published: February 2013

Background:  Islets obtained from genetically‐engineered (GE) pigs with Gal‐knockout (GTKO) and high expression of hCD46 have shown long‐term function and successful correction of insulin independence in nonhuman primates (NHP). Two anti‐coagulant genes (hTFPI and hCD39), and an immunosuppressive gene (pCTLA4Ig) have been added to the GTKO/hCD46 background, using an islet‐specific expression system, for inhibition of thrombosis, inflammation, early islet loss, and rejection. Transplant outcomes are being tested in a diabetic monkey model using adult islets from these multi‐transgenic pigs.

Methods:  Three individual vectors (CD39ins, TFPIins and CTLA4Igins), each under control of the rat insulin II promoter and mouse PDX‐1 enhancer, were transfected alone or in combination into GTKO/hCD46 pig fibroblast cells. GTKO/hCD46/CD39ins, GTKO/hCD46/TFPIins/CTLA4Igins, and GTKO/hCD46/CD39ins/TFPIins/CTLA4Igins cell lines were used for nuclear transfer (NT) to produce pregnancies. Cloned pigs were analyzed for transgene expression in pancreas, heart, and liver by IHC, Western, and real‐time PCR. Glucose metabolism in the various GE pigs was determined by measurement of blood glucose, c‐peptide, and via intravenous glucose tolerance tests (ivGTT) or arginine stimulation test. GE islets from adult pigs were transplanted intraportally into STZ‐diabetic monkeys using an ATG/MMF/anti‐CD154 immunosuppression regimen.

Results:  Viable multi‐transgenic pigs with three, four, or five different genetic modifications were produced. Three different pig lines were established: (i) GTKO/hCD46/CD39ins, (ii) GTKO/hCD46/TFPIins/CTLA4Igins, and (iii) GTKO/hCD46/CD39ins/TFPIins/CTLA4Igins. IHC revealed robust islet‐specific expression of CD39, TFPI and CTLA4Ig in pancreas, with only background expression in heart and liver. Western analysis showed strong pCTLA4Ig expression only in the pancreas. Blood glucose metabolism was normal in all GE pigs tested. Islets from GE pigs containing the TFPI transgene, when mixed with human blood, showed prolonged clotting times. Early islet loss (IBMIR) was reduced 5–7 fold in NHP transplants using islets from 4‐GE and 5‐GE pigs vs. 2‐GE pig islets.

Conclusions:  (i) Healthy multi‐transgenic pigs with islet‐specific expression of hCD39, hTFPI, and pCTLA4Ig were produced. (ii) Glucose metabolism was normal in these GE pigs. (iii) GE islets showed anti‐coagulation activity in vitro, and protection from IBMIR in vivo in NHP transplants. (iv) GE islets transplanted into diabetic monkeys demonstrated prolonged survival, function, and complete normalization of blood glucose levels for up to 1 year.
The potential of local immunomodulation and tolerance induction in porcine islet xenotransplantation
Authors: van Bürck Lelia , Seissler Jochen
Published: February 2013

Transplantation of pancreas or isolated islet cells is currently the only option to cure type 1 diabetes. The success of islet transplantation is still limited by the requirement of large numbers of high quality islets and the shortage of organ donors. Porcine islets are a promising cell source, but the intensive immunosuppressive regimen required to suppress rejection prevents the translation into clinical practice.

We aimed to develop a novel method to inhibit the human‐anti‐pig immune reaction by the expression of immunomodulatory molecules in porcine beta cells. Thus, a transgenic pig was generated expressing LEA29Y – a second generation human CTLA4‐Ig fusion protein, which inhibits activation of T cells by CD80/CD86‐CD28 costimulation – under the control of the porcine insulin promotor. Islet‐like clusters (ICC) from neonatal pigs were isolated and transplanted under the kidney capsule of diabetic NOD‐scid‐IL2γnull (NSG) mice. After an in vivo maturation period mice transplanted with wildtype (wt) as well as with LEA29Y transgenic (tg) ICCs developed normal glucose homeostasis. Within 30 days after the transfer of human PBMCs 80% of NSG mice transplanted with wt‐ICCs developed diabetes indicating xenograft rejection. By contrast, LEA‐tg ICCs were completely protected from rejection in all animals (1). Immunohistochemistry revealed a massive intra‐islet T cell infiltration, which was absent in the LEA‐tg ICCs. This proof of principle study suggests that specific expression of immunomodulatory molecules in beta cells does not disturb beta cell function and may have the potential to modulate immune response locally at the transplantation site without systemic immunosuppression.

To overcome the strong xenogeneic barrier of the human and cellular immune system a combination of LEA29Y with additional immunomodulatory factors may be required. Recently, Yi and coworkers demonstrated that the treatment with in vitro expanded regulatory T cells (Treg) prevents porcine islet rejection in humanized NSG mice by the suppression of the T cell‐mediated graft destruction (2). Other potential candidates to induce a state of tolerance against porcine islets currently under investigation are molecules targeting innate immunity and factors that prevent the reoccurrence of autoimmunity. Recent advances in xenotransplantation suggest that it may be possible to start with clinical trials using porcine neonatal or adult islets within the near future.

References:  1. KLYMIUK N, VAN BÜRCK L, BÄHR Aet al. Xenografted islet‐cell‐clusters from INSLEA29Y transgenic pigs rescue diabetes and prevent immune rejection in humanized mice. Diabetes 2012; 61:1527–1532.

2. YI S, JI M, WU J et al. Adoptive transfer with in vitro expanded human regulatory T cells protects against porcine islet xenograft rejection via interleukin‐10 in humanized mice. Diabetes 2012; 61:1180–1191.
Friction Resistance of Refined, Bleached and Deodorized (RBD) Palm Olein Using Modified Pin-on-disk Tribotester
Authors: Izhan M. I. , Rafiq A. K. Mohammed , Syahrullail S.
Published: February 2013

Daya geseran untuk minyak sawit olein yang ditapis, diluntur dan dinyahbau (RBD olein) telah dinilai dengan menggunakan alat tribotester pin-atas-cakera. RBD olein telah dipilih kerana sifat-sifat unggul tribologi dan pengeluarannya yang besar di Malaysia. Dalam kajian ini, 5ml minyak RBD olein telah
digunakan pada cakera beralur. Pin yang mempunyai hujung rata telah digunakan sebagai bahan ujikaji. Cakera dan pin diperbuat daripada titanium alloy. Beban normal yang dikenakan pada pin ialah 4.96N dan 19.68N. Ujikaji telah dijalankan selama satu jam. Kadar haus pin dan pekali geseran telah dikira. Ujikaji yang sama telah dijalankan dengan menggunakan minyak hidraulik komersial dan minyak mineral jenis parafinik yang tidak mengandungi bahan tambah. Pekali geseran yang dihasilkan oleh pelinciran minyak
sawit RBD olein adalah terendah berbanding minyak mineral parafinik pada kedua-dua jenis beban(4.96N dan 19.68N). Bagaimanapun, minyak sawit RBD olein menunjukkan kadar haus yang tinggi berbanding minyak mineral parafinik pada beban normal 19.68N.

Kata kunci: Pin-atas-cakera; minyak sawit; palm olein; haus; pekali geseran

A friction resistance of refined, bleached and deodorized (RBD) palm olein was evaluated using a modified pin-on-disk tribotester. RBD palm olein was selected due to its superior tribological properties and large production in Malaysia. In this research, 5ml of RBD palm olein was applied on the rotating grooved-disk. The flat-ended pin was used as a specimen. The material was titanium for both pin and disk. The normal loads applied are 4.96N and 19.68N. The experiments were run for one hour. The wear rate of the pin and friction coefficient was calculated. Same experiments were conducted using commercial hydraulic oil and additive-free paraffinic mineral oil. The results were compared with RBD palm olein.
The friction coefficient with the lubrication of RBD palm olein was the lowest for 4.96N and 19.68N under normal load conditions. Wear rate obtained by RBD palm olein at normal load 4.96N was the lowest; however, RBD palm olein showed a high wear rate when high normal load (19.68N) was applied.

Keywords: Pin-on-disk; palm oil; palm olein; wear; friction coefficient
Treatment of Industrial Textile Wastewater Using Polyarcrylamide (PAM) and Polyaluminium Chloride (PAC)
Authors: Ngadi Norzita , Yahya Noor Yahida , Muhamad Nabilah
Published: February 2013

Dalam kajian ini, poliacrylamid (PAM) dan polialuminium klorida (PAC) telah digunakan sebagai bahan pengental untuk merawat air sisa industri tekstil. Eksperimen ini telah dijalankan dengan menggunakan ujian balang. Kesan dos, kelajuan percampuran dan masa pemendapan ke atas keberkesanan proses flokulasi telah dikaji. Air sisa yang terawat dianalisis untuk penyingkiran warna, pengurangan COD dan pengurangan kekeruhan. Keputusan yang diperolehi menunjukkan bahawa PAM adalah lebih baik daripada PAC dalam merawat air sisa tekstil. PAM mencatatkan penurunan parameter tertinggi, iaitu 6 NTU untuk kekeruhan, 744 mg / l untuk COD, dan scala kurang daripada 0.5 untuk warna. Prestasi tebaik PAM dicapai pada dos 0.07 g dan apabila proses flokulasi dijalankan pada 200 rpm kelajuan percampuran
dan 30 min masa pemendapan. Kajian ini juga mendapati bahawa parameter operasi yang dikaji (iaitu dos, kelajuan percampuran dan masa pemendapan) tidak banyak mempengaruhi ke atas penyingkiran warna, dan penurunan kekeruhan dan COD apabila PAC digunakan sebagai bahan pengental.

Kata kunci: Pengelompokan; poliacrylamid (PAM); polialuminium klorida (PAC); air sisa tekstil

In this study, polyacrylamide (PAM) and polyaluminum chloride (PAC) was used as a flocculant to treat industrial textile wastewater. The experiment was conducted using a Jar test experiment. The effect of dosage, mixing speed and settling time on the performance of the fflocculation process was investigated.
The treated textile wastewater was analyzed by its color removal, turbidity and COD reductions. The results obtained showed that PAM performed better in treating the textile wastewater compared to PAC.PAM recorded the highest reduction of parameters, which are 6 NTU for turbidity, 744 mg/l for COD, and scale less than 0.5 for colour. The best performance of PAM was achieved at dosage 0.07 g and when the flocculation process was conducted at 200 rpm of mixing speed and 30 min of settling time. It was also found that the investigated operating parameters (i.e. dosage, mixing speed and settling time) did not influence much on removal of color and reduction of turbidity and COD when PAC was used as flocculant.

Keywords: Flocculation; polyacrylamide (PAM); polyaluminum chloride (PAC); textile wastewater
Profil Personaliti dalam Kalangan Pensyarah UTM yang Cemerlang dalam Pengajaran dan Pembelajaran Berdasarkan Penilaian Pengajaran Pensyarah
Authors: Abd Ghani Faizah
Published: February 2013

This study aims to identify the personality profile among UTM lecturers who were excellent in teaching and learning based on the lecturers‟ teaching evaluation system (PPP) for semester 2 session 2007/2008 and semester 1 2008/2009. This study is corellational study and used the Big Five Personality Traits as
framework research. A total of 52 lecturers had been selected using purposive sampling method. NEO-PIR inventory set by McCrae and Costa (1992) and translated by Khairul Anwar (2000) had been used in this study. It shows a high level of reliability which demonstrated the alpha Cronbach value of .870 for Neuroticism, .804 for Extraversion, .790 for Openness to Experience, .682 for Agreeableness and .904 for Conscientiousness. Overall, the value of alpha Cronbach is .813. The data collected were analyzed using 'Statistical Packages for Social Sciences(SPSS v12 for windows). The result of the descriptive analysis showed that the majority of excellent lecturers have conscientiousness profile of personality (mean=182.57), followed by agreeableness (mean=175.75) and extraversion (mean=160.019). The finding also showed that there was no significant between the profile of personality and achievement among the excellent lecturers.

Keywords:Personality profile; lecturer; excellent; teaching and learning; teaching evaluation

Kajian ini bertujuan untuk mengkaji profil personaliti pensyarah UTM yang cemerlang dalam pengajaran dan pembelajaran berdasarkan Penilaian Pengajaran Pensyarah (PPP) bagi semester 2 sesi 2007/2008 dan semester 1 sesi 2008/2009. Kajian ini merupakan kajian korelasi dan menggunakan Teori Tret Personaliti
Big Five sebagai kerangka kajian. Seramai 52 orang pensyarah cemerlang dalam PPP dilibatkan sebagai responden. Persampelan bertujuan digunakan dalam kajian ini dan Soal selidik NEO-PI-R oleh McCrae and Costa (1992) yang diterjemahkan oleh Khairul Anwar (2000) digunakan sebagai instrumen kajian. Soal Selidik ini mempunyai nilai kebolehpercayaan yang baik berdasarkan nilai alpha Cronbach .870 bagi domain Neuroticism (N), .804 bagi domain Extraversion (E), .790 bagi domain Openness to Experience (O), .682 bagi domain Agreeableness (A), dan .904 bagi domain Conscientiousness (C). Nilai alpha Cronbach bagi keseluruhan item adalah .813. Data yang telah dikumpul dianalisis menggunakan „Statistical Packages for Social Sciences’ (SPSS) versi 12.0 for windows. Analisis deskriptif menunjukkan bahawa majoriti pensyarah yang cemerlang memiliki profil personaliti conscientiousness
(min=182.57) diikuti dengan agreeableness (min=175.75) dan extraversion (min=160.019). Dapatan kajian menunjukkan tidak terdapat hubungan profil personaliti dengan pencapaian pensyarah yang cemerlang.

Kata kunci: Profil personaliti; pensyarah; cemerlang; pengajaran dan pembelajaran; penilaian pengajaran
Skeletal muscle mitochondrial dysfunction during chronic obstructive pulmonary disease: central actor and therapeutic target
Authors: Meyer Alain , Zoll Joffrey , Charles Anne Laure , Charloux Anne , de Blay Frédéric , Diemunsch Pierre , Sibilia Jean , Piquard François , Geny Bernard
Published: February 2013

New Findings

What is the topic of this review?

Muscle dysfunction is a common complication and an important independent prognostic factor in chronic obstructive pulmonary disease (COPD).

What advances does it highlight?

In COPD patients, the vastus lateralis muscle presents with alterations that include a decrease in mitochondrial density and biogenesis, impaired mitochondrial respiration and coupling, as well as increased mitochondrial production of reactive oxygen species, possibly associated with increased mitochondrial apoptosis. These mitochondrial changes are accessible to conventional therapies, such as exercise and tobacco cessation, but potentially also to innovative management strategies, such as antioxidant treatment and supplementation with polyunsaturated fatty acids. Mitochondrial pathophysiology represents an emerging area of research in muscle dysfunction associated with COPD and has promising therapeutic implications.

Muscle dysfunction is a common complication and an important prognostic factor in chronic obstructive pulmonary disease (COPD). As therapeutic strategies are still needed to treat this complication, gaining more insight into the process that leads to skeletal muscle decline in COPD appears to be an important issue. This review focuses on mitochondrial involvement in limb skeletal muscle alterations (decreased muscle mass, strength, endurance and power and increased fatigue) in COPD. Mitochondria are the main source of energy for the cells; they are involved in production of reactive oxygen species and activate an important pathway that leads to apoptosis. In COPD patients, skeletal muscles are characterized by decreased mitochondrial density and biogenesis, impaired activity and coupling of mitochondrial respiratory chain complexes, increased mitochondrial production of reactive oxygen species and, possibly, increased apoptosis. Of particular interest, a sedentary lifestyle, hypoxia, hypercapnia, tobacco smoking, corticosteroid therapy and, possibly, inflammation participate in this mitochondrial dysfunction, which is accessible to conventional therapies, such as exercise and tobacco cessation, as well as, potentially, to more innovative approaches, such as antioxidant treatment and supplementation with polyunsaturated fatty acids.
Blocking NRG1 and Other Ligand-Mediated Her4 Signaling Enhances the Magnitude and Duration of the Chemotherapeutic Response of Non–Small Cell Lung Cancer
Authors: Hegde Ganapati V. , de la Cruz Cecile C. , Chiu Cecilia , Alag Navneet , Schaefer Gabriele , Crocker Lisa , Ross Sarajane , Goldenberg David , Merchant Mark , Tien Janet , Shao Lily , Roth Leslie , Tsai Siao-Ping , Stawicki Scott , Jin Zhaoyu , Wyatt Shelby K. , Carano Richard A. D. , Zheng Yanyan , Sweet-Cordero E. Alejandro , Wu Yan , Jackson Erica L.
Published: February 2013

Inhibition of Her4 signaling enhances the response to chemotherapy and delays tumor regrowth after cessation of treatment.
Dual Inactivation of Akt and ERK by TIC10 Signals Foxo3a Nuclear Translocation, TRAIL Gene Induction, and Potent Antitumor Effects
Authors: Allen Joshua E. , Krigsfeld Gabriel , Mayes Patrick A. , Patel Luv , Dicker David T. , Patel Akshal S. , Dolloff Nathan G. , Messaris Evangelos , Scata Kimberly A. , Wang Wenge , Zhou Jun-Ying , Wu Gen Sheng , El-Deiry Wafik S.
Published: February 2013

TIC10 is a small molecule that activates Foxo3a through dual inactivation of Akt and ERK, up-regulates the expression of the TRAIL gene, an endogenous tumor suppressor, and effectively improves the therapeutic properties and utility of TRAIL as an anticancer therapy.
Targeting Protein Prenylation in Progeria
Authors: Young Stephen G. , Yang Shao H. , Davies Brandon S. J. , Jung Hea-Jin , Fong Loren G.
Published: February 2013

Results of a recent clinical trial in children with Hutchinson-Gilford progeria spur scientists to consider new therapeutic strategies.
Hha Controls Escherichia coli O157:H7 Biofilm Formation by Differential Regulation of Global Transcriptional Regulators FlhDC and CsgD
Authors: Sharma Vijay K. , Bearson Bradley L.
Published: February 2013

Although molecular mechanisms promoting adherence of enterohemorrhagic
Escherichia coli
(EHEC) O157:H7 on epithelial cells are well characterized, regulatory mechanisms controlling biofilm formation are not fully understood. In this study, we demonstrate that biofilm formation in EHEC O157:H7 strain 86-24 is highly repressed compared to that in an isogenic
hha
mutant. The
hha
mutant produced large quantities of biofilm compared to the wild-type strain at 30ï¿½C and 37ï¿½C. Complementation of the
hha
mutant reduced the level of biofilm formation to that of the wild-type strain, indicating that Hha is a negative regulator of biofilm production. While swimming motility and expression of the flagellar gene
fliC
were significantly reduced, the expression of
csgA
(encoding curlin of curli fimbriae) and the ability to bind Congo red were significantly enhanced. The expression of both
fliC
and
csgA
and the phenotypes of motility and curli production affected by these two genes, respectively, were restored to wild-type levels in the complemented
hha
mutant. The
csgA
deletion abolished biofilm formation in the
hha
mutant and wild-type strain, and
csgA
complementation restored biofilm formation to these strains, indicating the importance of
csgA
and curli in biofilm formation. The regulatory effects of Hha on flagellar and curli gene expression appear to occur via the induction and repression of FlhDC and CsgD, as demonstrated by reduced
flhD
and increased
csgD
transcription in the
hha
mutant, respectively. In gel shift assays Hha interacted with
flhDC
and
csgD
promoters. In conclusion, Hha regulates biofilm formation in EHEC O157:H7 by differential regulation of FlhDC and CsgD, the global regulators of motility and curli production, respectively.
Genetic Relationships of Vibrio parahaemolyticus Isolates from Clinical, Human Carrier, and Environmental Sources in Thailand, Determined by Multilocus Sequence Analysis
Authors: Theethakaew Chonchanok , Feil Edward J. , Castillo-Ramï¿½rez Santiago , Aanensen David M. , Suthienkul Orasa , Neil Douglas M. , Davies Robert L.
Published: February 2013

Vibrio parahaemolyticus
is a seafood-borne pathogenic bacterium that is a major cause of gastroenteritis worldwide. We investigated the genetic and evolutionary relationships of 101
V. parahaemolyticus
isolates originating from clinical, human carrier, and various environmental and seafood production sources in Thailand using multilocus sequence analysis. The isolates were recovered from clinical samples (
n
= 15), healthy human carriers (
n
= 18), various types of fresh seafood (
n
= 18), frozen shrimp (
n
= 16), fresh-farmed shrimp tissue (
n
= 18), and shrimp farm water (
n
= 16). Phylogenetic analysis revealed a high degree of genetic diversity within the
V. parahaemolyticus
population, although isolates recovered from clinical samples and from farmed shrimp and water samples represented distinct clusters. The tight clustering of the clinical isolates suggests that disease-causing isolates are not a random sample of the environmental reservoir, although the source of infection remains unclear. Extensive serotypic diversity occurred among isolates representing the same sequence types and recovered from the same source at the same time. These findings suggest that the O- and K-antigen-encoding loci are subject to exceptionally high rates of recombination. There was also strong evidence of interspecies horizontal gene transfer and intragenic recombination involving the
recA
locus in a large proportion of isolates. As the majority of the intragenic recombinational exchanges involving
recA
occurred among clinical and carrier isolates, it is possible that the human intestinal tract serves as a potential reservoir of donor and recipient strains that is promoting horizontal DNA transfer, driving evolutionary change, and leading to the emergence of new, potentially pathogenic strains.
Phylogenetic Delineation of the Novel Phylum Armatimonadetes (Former Candidate Division OP10) and Definition of Two Novel Candidate Divisions
Authors: Lee K. C. Y. , Herbold C. W. , Dunfield P. F. , Morgan X. C. , McDonald I. R. , Stott M. B.
Published: February 2013

Small-subunit (SSU) rRNA gene sequences associated with the phylum
Armatimonadetes
were analyzed using multiple phylogenetic methods, clarifying both the phylum boundary and the affiliation of previously ambiguous groupings. Here we define the
Armatimonadetes
as 10 class-level groups and reclassify two previously associated groups as candidate divisions WS1 and FBP.
Evaluating the Pathogenic Potential of Environmental Escherichia coli by Using the Caenorhabditis elegans Infection Model
Authors: Merkx-Jacques Alexandra , Coors Anja , Brousseau Roland , Masson Luke , Mazza Alberto , Tien Yuan-Ching , Topp Edward
Published: February 2013

The detection and abundance of
Escherichia coli
in water is used to monitor and mandate the quality of drinking and recreational water. Distinguishing commensal waterborne
E. coli
isolates from those that cause diarrhea or extraintestinal disease in humans is important for quantifying human health risk. A DNA microarray was used to evaluate the distribution of virulence genes in 148
E. coli
environmental isolates from a watershed in eastern Ontario, Canada, and in eight clinical isolates. Their pathogenic potential was evaluated with
Caenorhabditis elegans
, and the concordance between the bioassay result and the pathotype deduced by genotyping was explored. Isolates identified as potentially pathogenic on the basis of their complement of virulence genes were significantly more likely to be pathogenic to
C. elegans
than those determined to be potentially nonpathogenic. A number of isolates that were identified as nonpathogenic on the basis of genotyping were pathogenic in the infection assay, suggesting that genotyping did not capture all potentially pathogenic types. The detection of the adhesin-encoding genes
sfaD
,
focA
, and
focG
, which encode adhesins; of
iroN
2
, which encodes a siderophore receptor; of
pic
, which encodes an autotransporter protein; and of
b1432
, which encodes a putative transposase, was significantly associated with pathogenicity in the infection assay. Overall,
E. coli
isolates predicted to be pathogenic on the basis of genotyping were indeed so in the
C. elegans
infection assay. Furthermore, the detection of
C. elegans
-infective environmental isolates predicted to be nonpathogenic on the basis of genotyping suggests that there are hitherto-unrecognized virulence factors or combinations thereof that are important in the establishment of infection.
Changes in Enterococcal Populations and Related Antibiotic Resistance along a Medical Center-Wastewater Treatment Plant-River Continuum
Authors: Leclercq Roland , Oberlï¿½ Kenny , Galopin Sï¿½bastien , Cattoir Vincent , Budzinski Hï¿½lï¿½ne , Petit Fabienne
Published: February 2013

To determine if hospital effluent input has an ecological impact on downstream aquatic environment, antibiotic resistance in
Enterococcus
spp. along a medical center-retirement home-wastewater treatment plant-river continuum in France was determined using a culture-based method. Data on antibiotic consumption among hospitalized and general populations and levels of water contamination by antibiotics were collected. All isolated enterococci were genotypically identified to the species level, tested for
in vitro
antibiotic susceptibility, and typed by multilocus sequence typing. The
erm
(B) and
mef
(A) (macrolide resistance) and
tet
(M) (tetracycline resistance) genes were detected by PCR. Along the continuum, from 89 to 98% of enterococci, according to the sampled site, were identified as
Enterococcus faecium
. All
E. faecium
isolates from hospital and retirement home effluents were multiply resistant to antibiotics, contained
erm
(B) and
mef
(A) genes, and belonged to hospital-adapted clonal complex 17 (CC17). Even though this species remained dominant in the downstream continuum, the relative proportion of CC17 isolates progressively decreased in favor of other subpopulations of
E. faecium
that were more diverse, less resistant to antibiotics, and devoid of the classical macrolide resistance genes and that belonged to various sequence types. Antibiotic concentrations in waters were far below the MICs for susceptible isolates. CC17
E. faecium
was probably selected in the gastrointestinal tract of patients under the pressure of administered antibiotics and then excreted together with the resistance genes in waters to progressively decrease along the continuum.
Rewiring Lactococcus lactis for Ethanol Production
Authors: Solem Christian , Dehli Tore , Jensen Peter Ruhdal
Published: February 2013

Lactic acid bacteria (LAB) are known for their high tolerance toward organic acids and alcohols (R. S. Gold, M. M. Meagher, R. Hutkins, and T. Conway, J. Ind. Microbiol.
10:
45–54, 1992) and could potentially serve as platform organisms for production of these compounds. In this study, we attempted to redirect the metabolism of LAB model organism
Lactococcus lactis
toward ethanol production. Codon-optimized
Zymomonas mobilis
pyruvate decarboxylase (PDC) was introduced and expressed from synthetic promoters in different strain backgrounds. In the wild-type
L. lactis
strain MG1363 growing on glucose, only small amounts of ethanol were obtained after introducing PDC, probably due to a low native alcohol dehydrogenase activity. When the same strains were grown on maltose, ethanol was the major product and lesser amounts of lactate, formate, and acetate were formed. Inactivating the lactate dehydrogenase genes
ldhX
,
ldhB
, and
ldh
and introducing codon-optimized
Z. mobilis
alcohol dehydrogenase (ADHB) in addition to PDC resulted in high-yield ethanol formation when strains were grown on glucose, with only minor amounts of by-products formed. Finally, a strain with ethanol as the sole observed fermentation product was obtained by further inactivating the phosphotransacetylase (PTA) and the native alcohol dehydrogenase (ADHE).
Redundancy among Manganese Peroxidases in Pleurotus ostreatus
Authors: Salame Tomer M. , Knop Doriv , Levinson Dana , Yarden Oded , Hadar Yitzhak
Published: February 2013

Manganese peroxidases (MnPs) are key players in the ligninolytic system of white rot fungi. InPleurotus ostreatus(the oyster mushroom) these enzymes are encoded by a gene family comprising nine members,mnp1to -9(mnpgenes). Mn2+amendment toP. ostreatuscultures results in enhanced degradation of recalcitrant compounds (such as the azo dye orange II) and lignin. In Mn2+-amended glucose-peptone medium,mnp3,mnp4, andmnp9were the most highly expressedmnpgenes. After 7 days of incubation, the time point at which the greatest capacity for orange II decolorization was observed,mnp3expression and the presence of MnP3 in the extracellular culture fluids were predominant. To determine the significance of MnP3 for ligninolytic functionality in Mn2+-sufficient cultures,mnp3was inactivated via the Δku80strain-basedP. ostreatusgene-targeting system. In Mn2+-sufficient medium, inactivation ofmnp3did not significantly affect expression of nontargeted MnPs or their genes, nor did it considerably diminish the fungal Mn2+-mediated orange II decolorization capacity, despite the significant reduction in total MnP activity. Similarly, inactivation of eithermnp4ormnp9did not affect orange II decolorization ability. These results indicate functional redundancy within theP. ostreatusMnP gene family, enabling compensation upon deficiency of one of its members.
Evidence from Serpula lacrymans that 2,5-Dimethoxyhydroquinone Is a Lignocellulolytic Agent of Divergent Brown Rot Basidiomycetes
Authors: Korripally Premsagar , Timokhin Vitaliy I. , Houtman Carl J. , Mozuch Michael D. , Hammel Kenneth E.
Published: February 2013

Basidiomycetes that cause brown rot of wood are essential biomass recyclers in coniferous forest ecosystems and a major cause of failure in wooden structures. Recent work indicates that distinct lineages of brown rot fungi have arisen independently from ligninolytic white rot ancestors via loss of lignocellulolytic enzymes. Brown rot thus proceeds without significant lignin removal, apparently beginning instead with oxidative attack on wood polymers by Fenton reagent produced when fungal hydroquinones or catechols reduce Fe
3+
in colonized wood. Since there is little evidence that white rot fungi produce these metabolites, one question is the extent to which independent lineages of brown rot fungi may have evolved different Fe
3+
reductants. Recently, the catechol variegatic acid was proposed to drive Fenton chemistry in
Serpula lacrymans
, a brown rot member of the Boletales (D. C. Eastwood et al., Science 333:762-765, 2011). We found no variegatic acid in wood undergoing decay by
S. lacrymans
. We found also that variegatic acid failed to reduce
in vitro
the Fe
3+
oxalate chelates that predominate in brown-rotting wood and that it did not drive Fenton chemistry
in vitro
under physiological conditions. Instead, the decaying wood contained physiologically significant levels of 2,5-dimethoxyhydroquinone, a reductant with a demonstrated biodegradative role when wood is attacked by certain brown rot fungi in two other divergent lineages, the Gloeophyllales and Polyporales. Our results suggest that the pathway for 2,5-dimethoxyhydroquinone biosynthesis may have been present in ancestral white rot basidiomycetes but do not rule out the possibility that it appeared multiple times via convergent evolution.
Global Small RNA Chaperone Hfq and Regulatory Small RNAs Are Important Virulence Regulators in Erwinia amylovora
Authors: Zeng Quan , McNally R. Ryan , Sundin George W.
Published: February 2013

Hfq is a global small RNA (sRNA) chaperone that interacts with Hfq-regulated sRNAs and functions in the posttranscriptional regulation of gene expression. In this work, we identified Hfq to be a virulence regulator in the Gram-negative fire blight pathogen
Erwinia amylovora
. Deletion of
hfq
in
E. amylovora
Ea1189 significantly reduced bacterial virulence in both immature pear fruits and apple shoots. Analysis of virulence determinants in strain Ea1189Δ
hfq
showed that Hfq exerts pleiotropic regulation of amylovoran exopolysaccharide production, biofilm formation, motility, and the type III secretion system (T3SS). Further characterization of biofilm regulation by Hfq demonstrated that Hfq limits bacterial attachment to solid surfaces while promoting biofilm maturation. Characterization of T3SS regulation by Hfq revealed that Hfq positively regulates the translocation and secretion of the major type III effector DspE and negatively controls the secretion of the putative translocator HrpK and the type III effector Eop1. Lastly, 10 Hfq-regulated sRNAs were identified using a computational method, and two of these sRNAs, RprA and RyhA, were found to be required for the full virulence of
E. amylovora
.
Functional Analysis of the Protein Veg, Which Stimulates Biofilm Formation in Bacillus subtilis
Authors: Lei Ying , Oshima Taku , Ogasawara Naotake , Ishikawa Shu
Published: February 2013

Biofilm is a complex aggregate of cells that adhere to each other and produce an extracellular matrix. In
Bacillus subtilis
, an extracellular polysaccharide (EPS) and amyloid fiber (TasA), synthesized by the
epsA
-
epsO
and
tapA
-
sipW
-
tasA
operons, respectively, are the primary components of the extracellular matrix. In the current study, we investigated the functional role of the previously uncharacterized
veg
gene in
B. subtilis
. Overproduction of Veg, a small protein highly conserved among Gram-positive bacteria, stimulated biofilm formation via inducing transcription of the
tapA
-
sipW
-
tasA
operon. Moreover, overproduced Veg restored the impairment of biofilm formation in mutants carrying a deletion of of
sinI
,
slrA
, or
slrR
, encoding an antirepressor of SinR that acts as the master regulator of biofilm formation, while biofilm morphology in the absence of SinR was not affected by either additional
veg
deletion or overproduction, indicating that Veg negatively regulates SinR activity independently of the known antirepressors. Expression of
sinR
was not affected in Veg-overproducing cells, and amounts of SinR were similar in cells expressing different levels of Veg, strongly suggesting that Veg modulates the repressor activity of SinR. Interestingly, the results of
in vivo
pulldown assays of the SinR complex indicate that Veg inhibits the interactions between SinR and SlrR. Based on these findings, we propose that Veg or a Veg-induced protein acts as an antirepressor of SinR to regulate biofilm formation.
Dectin-1 Is Required for Resistance to Coccidioidomycosis in Mice
Authors: Viriyakosol Suganya , Jimenez Maria del Pilar , Gurney Michael A. , Ashbaugh Mark E. , Fierer Joshua
Published: February 2013

We assessed the role of Dectin-1 in the immune response to the pathogenic fungusCoccidioides, bothin vitroandin vivo, using mice with a targeted mutation inClec7a. Elicited peritoneal macrophages responded to formalin-killed spherules (FKS) and alkali-treated FKS by secreting proinflammatory cytokines in a Dectin-1- and β-glucan-dependent manner. The responses of bone marrow-derived dendritic cells (BMDC) to the same stimulants were more complex; interleukin 1β (IL-1β) and tumor necrosis factor alpha (TNF-α) secretion was independent of Dectin-1, while IL-6, IL-10, and granulocyte-macrophage colony-stimulating factor (GM-CSF) were largely but not entirely dependent on Dectin-1. After intranasal infection, Dectin-1−/−mice had lower concentrations of IL-12p70, gamma interferon (IFN-γ), IL-1β, and the Th17 cytokines IL-22, IL-23, and 17A in the lung lavage fluid, which may explain why they were significantly more susceptible to pulmonary coccidioidomycosis two weeks after infection. The Dectin-1 mutation was even more deleterious in (B6 × DBA/2)F2mice, which are more resistant to coccidioidomycosis than B6 mice by virtue of protective genes from DBA/2, a genetically resistant strain. We also found that two susceptible strains of mice (B6 and BALB/c) expressed much less Dectin-1 in their lungs than did resistant DBA/2 mice. We conclude that Dectin-1 is necessary for resistance toCoccidioides immitis, that Dectin-1 promotes both Th1 and Th17 protective immune responses to this infection, and that there is a correlation between expression of Dectin-1 by the inflammatory infiltrate and resistance to coccidioidomycosis.

IMPORTANCECoccidioidomycosis is a fungal infection endemic in the southwestern United States and neighboring Mexico, causing ~150,000 lung infections in people and resulting in ~17,000 hospitalizations annually in California alone. Very little is known about innate immunity to this fungus. This paper shows that Dectin-1, the primary β-glucan receptor on myeloid cells, is required for resistance to this pathogen. Dectin-1 is part of the innate immune system, and it is needed to direct the acquired immune response toward into a pathway that will lead to macrophage activation. Lungs from infected mice lacking Dectin-1 had lower concentrations of Th1 and Th17 cytokines, two cytokine pathways that are very important for acquired T cell immunity toCoccidioidesspp. This is the first demonstration that Dectin-1 is required for host resistance to a dimorphic, primary pathogenic fungus.
Perceptual Motor Features of Expert Acupuncture Lifting-Thrusting Skills
Authors: Li Jing , Grierson Lawrence E M , Wu Mary X , Breuer Ronny , Carnahan Heather
Published: February 2013

Background

Little is known with regard to how acupuncture skills are optimally taught, learnt and evaluated despite clear evidence that technical skill acquisition is important to trainee success in health professions.

Objectives

This study reports an investigation of the sensorimotor aspects of the acupuncture lifting-thrusting skill in order to highlight the important kinematic and kinetic features of the action. The study also explores the role of perceptual acuity in accurate acupuncture performance.

Methods

Twelve novice and 12 expert acupuncturists watched a standardised video demonstrating the mild reinforcing and reducing technique of lifting-thrusting on an acupuncture point and then performed 10 trials of the technique on an artificial skin pad mounted on a six-axis force transducer with an infrared light-emitting diode affixed to the index finger of their dominant hand. The force transducer measured the force applied by participants as they needled the acupuncture point while an optoelectric camera measured the position of the diode. Subsequently, the participants engaged in two tests of general perceptual acuity.

Results

Repeated measures analyses of variance indicated that experts are more consistent in their trial-by-trial amplitude (p=0.03) and lifting-thrusting velocity (p=0.029) than novices. Measures of perceptual acuity revealed no differences between novices and experts.

Conclusions

Movement amplitude and velocity consistency are the action features of the mild reinforcing and reducing lifting-thrusting skill that differentiate the performances of experts from novices. The acquisition of acupuncture expertise is a function of extended practice rather than any inherent perceptual ability.