bluuebunny/crossref_metadata_2025 huggingface dataset:
Overcoming burdens in the regulation of clinical research in children. Proceedings of a consensus conference, in historical context
Authors: Levine Robert J , Genel Myron , Cuttler Leona , Becker Dorothy J , Nieman Lynnette , Rosenfield Robert L
Published: December 2011

Background

Many investigators are concerned that the modes of implementation and enforcement of the federal regulations designed to protect children are unduly impeding pediatric clinical research.

Objective

To assess regulatory impediments to clinical research involving children and to develop recommendations to ameliorate them.

Participants

The Pediatric Endocrine Society and The Endocrine Society convened a consensus conference involving experts and stakeholders in patient-oriented research involving children and adolescents in 2008.

Consensus process

Following presentations that reviewed problematic issues around key regulations, participants divided into working groups to develop potential solutions that could be adopted at local and federal levels. Presentations to the full assembly were then debated. A writing committee then drafted a summary of the discussions and main conclusions, placing them in historical context, and submitted it to all participants for comment with the aim of developing consensus.

Conclusions

Recommendations designed to facilitate the ethical conduct of research involving children addressed the interpretation of ambiguous regulatory terms such as "minimal risk" and "condition" and called for the development by professional societies of best practice primers for common research procedures that would be informative to both investigators and institutional review boards. A call was issued for improved guidance from the Office for Human Research Protections and Food and Drug Administration as well as for the development by professional societies of a process to monitor progress in improving human subject research regulation. Finally, a need for systematic research to define the nature and extent of institutional obstacles to pediatric research was recognized.
Antibody response of healthy children to pandemic A/H1N1/2009 influenza virus
Authors: Esposito Susanna , Daleno Cristina , Tagliabue Claudia , Scala Alessia , Picciolli Irene , Taroni Francesca , Galeone Carlotta , Baldanti Fausto , Principi Nicola
Published: December 2011

Background

Little is known about the proportion of pediatric pandemic A/H1N1/2009 influenza cases who showed seroconversion, the magnitude of this seroconversion, or the factors that can affect the antibody level evoked by the pandemic A/H1N1/2009 influenza. Aims of this study were to analyse antibody responses and the factors associated with high antibody titres in a cohort of children with naturally acquired A/H1N1/2009 influenza infection confirmed by reverse-transcriptase polymerase chain reaction (RT-PCR).

Results

Demographic, clinical and virologic data were collected from 69 otherwise healthy children with pandemic A/H1N1/2009 influenza (27 females, mean age ± SD: 5.01 ± 4.55 years). Their antibody levels against pandemic A/H1N1/2009 and seasonal A/H1N1 influenza viruses were evaluated by measuring hemagglutination-inhibiting antibodies using standard assays. Sixty-four patients (92.8%) with pandemic A/H1N1/2009 influenza had A/H1N1/2009 antibody levels of ≥40, whereas only 28/69 (40.6%) were seroprotected against seasonal A/H1N1 influenza virus. Those who were seroprotected against seasonal A/H1N1 virus were significantly older, significantly more often hospitalised, had a diagnosis of pneumonia significantly more frequently, and were significantly more often treated with oseltamivir than those who were not seroprotected (p < 0.05). The children with the most severe disease (assessed on the basis of a need for hospitalisation and a diagnosis of pneumonia) had the highest antibody response against pandemic A/H1N1/2009 influenza virus.

Conclusions

Otherwise healthy children seem to show seroprotective antibody titres after natural infection with pandemic A/H1N1/2009 influenza virus. The strength of the immune response seems to be related to the severity of the disease, but not to previous seasonal A/H1N1 influenza immunity.
Primary pleural angiosarcoma as a mimicker of mesothelioma: a case report
Authors: Kao Yu-Chien , Chow Jyh-Ming , Wang Kum-Min , Fang Chia-Lang , Chu Jan-Show , Chen Chi-Long
Published: December 2011

Primary pleural angiosarcoma is a rare and clinically aggressive tumor. Patients usually present with chest pain, dyspnea, hemoptysis and/or cough. Radiologic studies reveal diffuse pleural thickening and pleural effusion with or without mass lesion. The clinical and radiological features both resemble those of mesothelioma, and its definite diagnosis requires careful histologic examination. However, frequent epithelioid feature and immunoreactivity to cytokeratin in primary pleural angiosarcoma further complicate the pathologic diagnosis. The use of proper immunohistochemical stains is often needed to support endothelial differentiation in the tumor cells and to exclude metastatic carcinoma and mesothelioma. We report the case of a 49-year-old male patient with primary pleural angiosarcoma, who presented with initial hemothorax, followed by a rapid progress to an inoperable status.

Virtual Slides

The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1059343251633446
An improved protocol for efficient transformation and regeneration of diverse indica rice cultivars
Authors: Sahoo Khirod K , Tripathi Amit K , Pareek Ashwani , Sopory Sudhir K , Singla-Pareek Sneh L
Published: December 2011

Background

Rice genome sequencing projects have generated remarkable amount of information about genes and genome architecture having tremendous potential to be utilized in both basic and applied research. Success in transgenics is paving the way for preparing a road map of functional genomics which is expected to correlate action of a gene to a trait in cellular and organismal context. However, the lack of a simple and efficient method for transformation and regeneration is a major constraint for such studies in this important cereal crop.

Results

In the present study, we have developed an easy, rapid and highly efficient transformation and regeneration protocol using mature seeds as explants and found its successful applicability to a choice of elite indica rice genotypes. We have optimized various steps of transformation and standardized different components of the regeneration medium including growth hormones and the gelling agent. The modified regeneration medium triggers production of large number of shoots from smaller number of calli and promotes their faster growth, hence significantly advantageous over the existing protocols where the regeneration step requires maximum time. Using this protocol, significantly higher transformation efficiency (up to 46%) and regeneration frequency (up to 92% for the untransformed calli and 59% for the transformed calli) were achieved for the four tested cultivars. We have used this protocol to produce hundreds of independent transgenic lines of different indica rice genotypes. Upon maturity, these transgenic lines were fertile thereby indicating that faster regeneration during tissue culture did not affect their reproductive potential.

Conclusions

This speedy, yet less labor-intensive, protocol overcomes major limitations associated with genetic manipulation in rice. Moreover, our protocol uses mature seeds as the explant, which can easily be obtained in quantity throughout the year and kept viable for a long time. Such an easy, efficient and generalized protocol has the potential to be a major tool for crop improvement and gene-function studies on the model monocot plant rice.
BPEX Pig Health Scheme: a useful monitoring system for respiratory disease control in pig farms?
Authors: Holt Hannah R , Alarcon Pablo , Velasova Martina , Pfeiffer Dirk U , Wieland Barbara
Published: December 2011

Background

Respiratory diseases account for significant economic losses to the UK pig industry. Lesions indicative of respiratory disease in pig lungs at slaughter e.g. pneumonia and pleuritis are frequently recorded to assess herd health or provide data for epidemiological studies. The BPEX Pig Health Scheme (BPHS) is a monitoring system, which informs producers of gross lesions in their pigs' carcasses at slaughter, enabling farm-level decisions to be made. The aim of the study was to assess whether information provided by the BPHS regarding respiratory lesions was associated with respiratory pathogens in the farm, farm management practices and each other.

Results

BPHS reports were obtained from a subset of 70 pig farms involved in a cross-sectional study conducted in 2008-09 investigating the epidemiology of post-weaning multi-systemic wasting syndrome. The reports were combined with data regarding the presence/absence of several pathogens in the herd and potential farm-level risk factors for respiratory disease. Principal component analysis (PCA) performed on BPHS reports generated three principal components, explaining 71% of the total variance. Enzootic pneumonia score, severe pleurisy and acute pleuropneumonia had the highest loadings for the principal component which explained the largest percentage of the total variance (35%) (BPHS component 1), it was thought that this component identifies farms with acute disease. Using the factor loadings a score for each farm for BPHS component 1 was obtained. As farms' score for BPHS component 1 increased, average carcass weight at slaughter decreased. In addition, farms positive for H1N2 and porcine reproductive and respiratory disease virus (PRRSV) were more likely to have higher levels of severe and mild pleurisy reported by the BPHS, respectively.

Conclusions

The study found statistical associations between levels of pleurisy recorded by BPHS at slaughter and the presence H1N2 and PRRSV in the herd. There is also some evidence that farms which submit pigs with these lesions may have reduced productivity. However, more research is needed to fully validate the scheme.
Congenital Dyserythropoietic Anemia Type II: molecular analysis and expression of the SEC23B Gene
Authors: Punzo Francesca , Bertoli-Avella Aida M , Scianguetta Saverio , Della Ragione Fulvio , Casale Maddalena , Ronzoni Luisa , Cappellini Maria D , Forni Gianluca , Oostra Ben A , Perrotta Silverio
Published: December 2011

Background

Congenital dyserythropoietic anemia type II (CDAII), the most common form of CDA, is an autosomal recessive condition. CDAII diagnosis is based on invasive, expensive, and time consuming tests that are available only in specialized laboratories. The recent identification ofSEC23Bmutations as the cause of CDAII opens new possibilities for the molecular diagnosis of the disease. The aim of this study was to characterize molecular genomicSEC23Bdefects in 16 unrelated patients affected by CDAII and correlate the identified genetic alterations withSEC23Btranscript and protein levels in erythroid precursors.

Methods

SEC23Bwas sequenced in 16 patients, their relatives and 100 control participants.SEC23Btranscript level were studied by quantitative PCR (qPCR) in peripheral erythroid precursors and lymphocytes from the patients and healthy control participants. Sec23B protein content was analyzed by immunoblotting in samples of erythroblast cells from CDAII patients and healthy controls.

Results

All of the investigated cases carriedSEC23Bmutations on both alleles, with the exception of two patients in which a single heterozygous mutation was found. We identified 15 differentSEC23Bmutations, of which four represent novel mutations: p.Gln214Stop, p.Thr485Ala, p.Val637Gly, and p.Ser727Phe. The CDAII patients exhibited a 40-60% decrease ofSEC23BmRNA levels in erythroid precursors when compared with the corresponding cell type from healthy participants. The largest decrease was observed in compound heterozygote patients with missense/nonsense mutations. In three patients, Sec23B protein levels were evaluated in erythroid precursors and found to be strictly correlated with the reduction observed at the transcript level. We also demonstrate that Sec23B mRNA expression levels in lymphocytes and erythroblasts are similar.

Conclusions

In this study, we identified four novelSEC23Bmutations associated with CDAII disease. We also demonstrate that the genetic alteration results in a significant decrease ofSEC23Btranscript in erythroid precursors. Similar down-regulation was observed in peripheral lymphocytes, suggesting that the use of these cells might be sufficient in the identification of Sec23B gene alterations. Finally, we demonstrate that decreased Sec23B protein levels in erythroid precursors correlate with down-regulation of theSEC23BmRNA transcript.
Centrosomal and mitotic abnormalities in cell lines derived from papillary thyroid cancer harboring specific gene alterations
Authors: Maric Irena , Viaggi Silvia , Caria Paola , Frau Daniela V , Degan Paolo , Vanni Roberta
Published: December 2011

Background

Differentiated thyroid carcinoma offers a good model to investigate the possible correlation between specific gene mutations and chromosome instability. Papillary thyroid neoplasms are characterized by different mutually exclusive genetic alterations, some of which are associated with aneuploidy and aggressive phenotype.

Results

We investigated the centrosome status and mitotic abnormalities in three thyroid carcinoma-derived cell lines, each maintaining the specific, biologically relevant gene alteration harbored by the parental tumors: RET/PTC1 rearrangement in TPC1; heterozygous and homozygous BRAF

V600E
mutation in K1 and in B-CPAP, respectively. B-CPAP cells showed a statistically significant (P < 0.01) higher frequency of abnormal mitotic figures compared to TPC1 and K1 cells.

Conclusions

Our data indicate that RET/PTC1 oncogenic activity is not related to mitotic chromosome impairment and missegregation whereas, based on the consistent difference in types/frequencies of centrosome and spindle abnormalities observed between K1 and B-CPAP cells, the hetero/homozygous allelic status of BRAF

V600E
mutation seems to be not irrelevant in respect to chromosomal instability development.
Comparison of RNA extraction methods from biofilm samples of Staphylococcus epidermidis
Authors: França Angela , Melo Luís DR , Cerca Nuno
Published: December 2011

Background

Microbial biofilms are communities of bacteria adhered to a surface and surrounded by an extracellular polymeric matrix. Biofilms have been associated with increased antibiotic resistance and tolerance to the immune system. Staphylococcus epidermidis is the major bacterial species found in biofilm-related infections on indwelling medical devices. Obtaining high quality mRNA from biofilms is crucial to validate the transcriptional measurements associated with the switching to the biofilm mode of growth. Therefore, we selected three commercially available RNA extraction kits with distinct characteristics, including those using silica membrane or organic extraction methods, and enzymatic or mechanical cell lysis, and evaluated the RNA quality obtained from two distinct S. epidermidis bacterial biofilms.

Results

RNA extracted using the different kits was evaluated for quantity, purity, integrity, and functionally. All kits were able to extract intact and functional total RNA from the biofilms generated from each S. epidermidis strain. The results demonstrated that the kit based on mechanical lysis and organic extraction (FastRNA® Pro Blue) was the only one that was able to isolate pure and large quantities of RNA. Normalized expression of the icaA virulence gene showed that RNA extracted with PureLink™ had a significant lower concentration of icaA mRNA transcripts than the other kits tested.

Conclusions

When working with complex samples, such as biofilms, that contain a high content extracellular polysaccharide and proteins, special care should be taken when selecting the appropriate RNA extraction system, in order to obtain accurate, reproducible, and biologically significant results. Among the RNA extraction kits tested, FastRNA® Pro Blue was the best option for both S. epidermidis biofilms used.
What works to meet the sexual and reproductive health needs of women living with HIV/AIDS
Authors: Gay Jill , Hardee Karen , Croce‐Galis Melanie , Hall Carolina
Published: December 2011

It is critical to include a sexual and reproductive health lens in HIV programming as most HIV transmission occurs through sexual intercourse. As global attention is focusing on the sexual and reproductive health needs of women living with HIV, identifying which interventions work becomes vitally important. What evidence exists to support sexual and reproductive health programming related to HIV programmes?

This article reviews the evidence of what works to meet the sexual and reproductive health needs of women living with HIV in developing countries and includes 35 studies and evaluations of eight general interventions using various methods of implementation science from 15 countries. Data are primarily from 2000‐2009. Searches to identify effective evaluations used SCOPUS, Popline, Medline, websites and consultations with experts. Evidence was ranked using the Gray Scale.

A range of successful and promising interventions to improve the sexual and reproductive health and rights of women living with HIV include: providing contraceptives and family planning counselling as part of HIV services; ensuring early postpartum visits providing family planning and HIV information and services; providing youth‐friendly services; supporting information and skills building; supporting disclosure; providing cervical cancer screening; and promoting condom use for dual protection against pregnancy and HIV. Provision of antiretrovirals can also increase protective behaviours, including condom use.

While many gaps in programming and research remain, much can be done now to operationalize evidence‐based effective interventions to meet the sexual and reproductive health needs of women living with HIV.
Disturbances of the hypothalamic-pituitary-adrenal axis and plasma electrolytes during experimental sepsis
Authors: Flierl Michael A , Rittirsch Daniel , Weckbach Sebastian , Huber-Lang Markus , Ipaktchi Kyros , Ward Peter A , Stahel Philip F
Published: December 2011

Background

Sepsis continues to be a poorly understood syndrome with a high mortality rate. While we are beginning to decipher the intricate interplay of the inflammatory response during sepsis, the precise regulation of the hypothalamic-pituitary-adrenal (HPA) axis and its impact on electrolyte homeostasis during sepsis remains incompletely understood.

Methods

Sepsis was induced in adult male Sprague-Dawley rats by cecal ligation and puncture (CLP). Plasma samples were obtained as a function of time (6-48 hrs) after CLP and compared with healthy animals (neg ctrl). Samples were analyzed for adrenocorticotropin (ACTH), corticosterone, and aldosterone levels, as well as concentrations of sodium (Na+), potassium (K+), chloride (Cl-), and magnesium (Mg2+).

Results

ACTH levels were found to be significantly reduced 6-24 hrs after CLP in comparison to baseline levels and displayed gradual recovery during the later course (24-48 hrs) of sepsis. Plasma corticosterone concentrations exhibited a bell-shaped response, peaking between 6 and 12 hrs followed by rapid decline and concentrations below negative control levels 48 hrs after injury. Aldosterone levels in septic animals were continuously elevated between 6 and 48 hrs. Whereas plasma Na+ levels were found to be persistently elevated following CLP, levels of K+, Cl- and Mg2+ were significantly reduced as a function of time and gradually recovered during the later course of sepsis.

Conclusions

CLP-induced sepsis resulted in dynamic changes of ACTH, corticosterone, and aldosterone levels. In addition, electrolyte levels showed significant disturbances after CLP. These electrolyte perturbations might be evoked by a downstream effect or a dysfunctional HPA-axis response during sepsis and contribute to severe complications during sepsis.
MicroRNAs and their isomiRs function cooperatively to target common biological pathways
Authors: Cloonan Nicole , Wani Shivangi , Xu Qinying , Gu Jian , Lea Kristi , Heater Sheila , Barbacioru Catalin , Steptoe Anita L , Martin Hilary C , Nourbakhsh Ehsan , Krishnan Keerthana , Gardiner Brooke , Wang Xiaohui , Nones Katia , Steen Jason A , Matigian Nicholas A , Wood David L , Kassahn Karin S , Waddell Nic , Shepherd Jill , Lee Clarence , Ichikawa Jeff , McKernan Kevin , Bramlett Kelli , Kuersten Scott , Grimmond Sean M
Published: December 2011

Background

Variants of microRNAs (miRNAs), called isomiRs, are commonly reported in deep-sequencing studies; however, the functional significance of these variants remains controversial. Observational studies show that isomiR patterns are non-random, hinting that these molecules could be regulated and therefore functional, although no conclusive biological role has been demonstrated for these molecules.

Results

To assess the biological relevance of isomiRs, we have performed ultra-deep miRNA-seq on ten adult human tissues, and created an analysis pipeline called miRNA-MATE to align, annotate, and analyze miRNAs and their isomiRs. We find that isomiRs share sequence and expression characteristics with canonical miRNAs, and are generally strongly correlated with canonical miRNA expression. A large proportion of isomiRs potentially derive from AGO2 cleavage independent of Dicer. We isolated polyribosome-associated mRNA, captured the mRNA-bound miRNAs, and found that isomiRs and canonical miRNAs are equally associated with translational machinery. Finally, we transfected cells with biotinylated RNA duplexes encoding isomiRs or their canonical counterparts and directly assayed their mRNA targets. These studies allow us to experimentally determine genome-wide mRNA targets, and these experiments showed substantial overlap in functional mRNA networks suppressed by both canonical miRNAs and their isomiRs.

Conclusions

Together, these results find isomiRs to be biologically relevant and functionally cooperative partners of canonical miRNAs that act coordinately to target pathways of functionally related genes. This work exposes the complexity of the miRNA-transcriptome, and helps explain a major miRNA paradox: how specific regulation of biological processes can occur when the specificity of miRNA targeting is mediated by only 6 to 11 nucleotides.
The RANKL Distal Control Region Is Required for the Increase in RANKL Expression, But Not the Bone Loss, Associated with Hyperparathyroidism or Lactation in Adult Mice
Authors: Onal Melda , Galli Carlo , Fu Qiang , Xiong Jinhu , Weinstein Robert S. , Manolagas Stavros C. , O'Brien Charles A.
Published: December 2011

Osteoclast-mediated bone resorption plays an essential role in calcium homeostasis and lactation. The cytokine receptor activator of nuclear factor κB ligand (RANKL) is one of a number of factors that controls the production, survival, and activity of osteoclasts. Calciotropic hormones, such as PTH, control RANKL transcription in part via an enhancer known as the distal control region (DCR), and mice lacking this enhancer have fewer osteoclasts under normal physiological conditions. Here, we have addressed the role of the DCR in situations in which activation of the PTH receptor is thought to stimulate bone resorption via elevation of RANKL expression. Dietary calcium deficiency stimulated RANKL expression in the bone of young (1 month old) wild-type, but not DCR knockout (KO), mice. Consistent with this, the cancellous bone loss and the increase in osteoclasts caused by dietary calcium deficiency were blunted in young KO mice. DCR deletion also prevented the increase in RANKL expression caused by dietary calcium deficiency in 6-month-old mice. However, the diet-induced bone loss was similar in wild-type and KO mice at this age. The increase in RANKL expression caused by lactation was also blunted in DCR KO mice, but lactation-induced bone loss was similar in both genotypes. These results demonstrate that, even though the DCR is required for the increase in RANKL expression associated with hyperparathyroidism or lactation, this increase is not required for the bone loss caused by these conditions in adult mice, suggesting that changes in other factors, such as osteoprotegerin or estrogen levels, play a dominant role.
Ambident Reactivity of Chloro(dialkylamino)(diphenylphosphinoyl)methanes
Authors: Morgalyuk Vasilii Petrovich , Strelkova Tat’yana Vladimirovna , Nifant’ev Eduard Eugenievich
Published: December 2011

The chemical properties of chloro(dialkylamino)(diphenylphosphinoyl)methanes have been studied using the simplest compound of this series, chloro(dimethylamino)(diphenylphosphinoyl)methane, as an example. Chloro(dimethylamino)(diphenylphosphinoyl)methane shows ambident reactivity when reacting with electrophiles and nucleophiles depending on coreactant nature, it behaves as either electrophilic substrate or phosphorus nucleophile. This fact can be explained by its dissociation in solutions with both C–Cl bond cleavage to give (dimethylamino)(diphenylphosphinoyl)methyl cation and Cl− anion and C–P bond cleavage to form chloro(dimethylamino)methyl cation and diphenylphosphinite anion. The capability of chloro(dimethylamino)(diphenylphosphinoyl)methane to produce spontaneously Ph2PO–anion allows us to recommend application in organic and organophosphorus synthesis as a synthetic equivalent (synthon) of diphenylphosphinite anion.
Synthesis and evaluation of novel phosphasugar anticancer agents
Authors: Yamaoka Mayumi , Yamashita Mitsuji , Yamada Manabu , Fujie Michio , Kiyofuji Keita , Ozaki Nobuhisa , Asai Kazuhide , Niimi Taishi , Suyama Takuya , Yamashita Junko , Sawada Akiho , Makita Reiko , Sugiyama Masaki , Toda Mitsuo , Nakamura Satoki , Ohnishi Kazunori
Published: December 2011

Starting materials of phosphasugars, 1-phenyl-2-phospholene 1-oxides, were prepared from dienes and phenylphosphonous dichloride (dichlorophenylphosphine). Several substituted novel phosphasugars (3- or 4-halo-substituted)-1-phenyl-2-phospholene 1-oxides as well as 1-phenyl-2-phospholane 1-oxides were prepared from 2-phospholenes. The synthesized compounds were evaluated for their antitumor activities against the leukemia cell lines (U937 and K562) by in vitro 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. 2,3,4-Tribromo-3-methyl-1-phenylphospholane 1-oxide showed superior antitumor activity against U937 and K562 cell lines in a comparative evaluation with Glivec. The analysis by flow cytometry implied that 2,3-dibromo-3-methyl-1-phenylphospholane 1-oxide induced apoptosis to leukemia cell lines.
The role of IgG avidity determination in diagnosis of Epstein-Barr virus infection in immunocompetent and immunocompromised patients
Authors: Vilibic-Cavlek Tatjana , Ljubin-Sternak Suncanica , Kos Ljiljana , Mlinaric-Galinovic Gordana
Published: December 2011

There is a high degree of variability in the serologic response to Epstein-Barr virus (EBV) infection, especially in viral capsid antigen (VCA)-IgM antibodies. Therefore, additional tests are needed to confirm primary infection.We evaluated the value of IgG avidity determination in diagnosis of EBV infection in immunocompetent and immunocompromised patients.A total of 236 serum samples from immunocompetent patients with symptoms suggestive of EBV infection were tested for the presence of VCA-IgM/IgG antibodies and IgG avidity. Using IgG avidity, acute primary infection was confirmed in 56.7% of the immunocompetent patients with positive and in 1.8% of patients with negative VCA-IgM. Recent primary infection was documented in 8.9% of the IgM positive and 3.5% of the IgM negative patients. In patients with indeterminate serology (equivocal IgM), 6.7% were classified by avidity index (AI) as acute primary infection, 10.0% as post-acute and 83.3% as past infection cases.Concerning the 32 immunocompromised patients, recent primary infection was documented in 3 of the 14 IgM positive patients. High AI was detected in 11 of these patients, indicating an IgM response due to reactivation.Determination of IgG avidity in combination with classical serologic markers seems to be a reliable method to confirm primary infection both in immunocompetent and immunocompromised patients. It may be especially useful to differentiate cases of primary infection in patients with undetectable VCA-IgM antibodies or indeterminate routine EBV serology.
Diversity of aminoglycoside modifying enzyme genes among multidrug resistant Acinetobacter baumannii genotypes isolated from nosocomial infections in Tehran hospitals and their association with class 1 integrons
Authors: Asadollahi Khairollah , Taherikalani Morovat , Maleki Abbas , Alizadeh Eshrat , Valadbaigi Hasan , Soroush Setareh , Maleki Hossain , Asadollahi Parisa , Emaneini Mohammad
Published: December 2011

The aim of the present study was to investigate, for the first time, the diversity of the genes encoding aminoglycoside-modifying enzymes (AME) and their association with class 1 integrons in IranianAcinetobacter baumanniistrains.A total of 100 multidrug resistantA. baumannii, isolated from eight distinct hospitals in Tehran, were enrolled in this study. Susceptibility of these isolates to antimicrobial agents including gentamicin and amikacin was determined by E-test. Aminoglycoside resistant isolates were then tested by PCR for AME genes, includingaphA6, aacC1, aacC2, aacA4, aadB, aadA1, classes 1 integron, 5′-CS-3′ and typed by RAPD PCR.The rate of resistance to imipenem, meropenem, gentamicin and amikacin were 39%, 39%, 38% and 32%, respectively. Intermediate resistance phenotype to gentamicin and amikacin was observed in 2% and 5% of all the isolates, respectively. Afteraph6 with 90% (n = 36/40),aadA1,aacC1 andaadB with 82.5% (n = 33/40), 65% (n = 26/40) and 20% (n = 8/40) were the most prevalent AME genes among aminoglycosides resistantA. baumanniiisolates. A combination of two to four different resistance genes was observed in 39 of 40 strains (97.5%), with a total of 7 different combinations. PCR of integrase genes revealed that AME gene was associated with 67% of class 1 integrons. RAPD analysis showed three predominant genotypes A (n = 20), B (n = 10) and 10 unrelated genotypes.The occurrence of identical resistance genes, gene combinations and class 1 integrons associated with these genes in clonally distinct strains indicates that horizontal gene transfer plays a major role in the dissemination of aminoglycoside resistance inA. baumannii.
Bacterial communities in an ultrapure water containing storage tank of a power plant
Authors: Bohus Veronika , Kéki Zsuzsa , Márialigeti Károly , Baranyi Krisztián , Patek Gábor , Schunk János , Tóth Erika
Published: December 2011

Ultrapure waters (UPWs) containing low levels of organic and inorganic compounds provide extreme environment. On contrary to that microbes occur in such waters and form biofilms on surfaces, thus may induce corrosion processes in many industrial applications. In our study, refined saltless water (UPW) produced for the boiler of a Hungarian power plant was examined before and after storage (sampling the inlet [TKE] and outlet [TKU] waters of a storage tank) with cultivation and culture independent methods. Our results showed increased CFU and direct cell counts after the storage. Cultivation results showed the dominance of aerobic, chemoorganotrophic α-Proteobacteria in both samples. In case of TKU sample, a more complex bacterial community structure could be detected. The applied molecular method (T-RFLP) indicated the presence of a complex microbial community structure with changes in the taxon composition: while in the inlet water sample (TKE) α-Proteobacteria (Sphingomonas sp., Novosphingobium hassiacum) dominated, in the outlet water sample (TKU) the bacterial community shifted towards the dominance of α-Proteobacteria (Rhodoferax sp., Polynucleobacter sp., Sterolibacter sp.), CFB (Bacteroidetes, formerly Cytophaga-Flavobacterium-Bacteroides group) and Firmicutes. This shift to the direction of fermentative communities suggests that storage could help the development of communities with an increased tendency toward corrosion.
Evaluation of mammalian codon usage of fimH in DNA vaccine design
Authors: Bagherpour Ghasem , Fooladi Abbas , Mehrabadi Jalil , Nourani Mohammad , Einollahi Behzad
Published: December 2011

UropathogenicEscherichia coli(UPEC) bacteria are the principal cause of urinary tract infections (UTI). Because these bacteria propagate intracellularly, the cellular immune response is an important factor in UTIs. Therefore, we designed a genetic construct to induce a cellular immune response. In order to develop a genetic construct that induces strong cellular immunity against this pathogen, we used thefimHsynthetic gene according to mammalian codon usage, and the gene expression was compared with wild type codon usage. Initially, we designed two constructs, pVAX/fimHmam and pVAX/fimHwt, which contain mammalian and wild type codon usage, respectively. The Cos-7 cell line was transfected separately with a complex of pVAX/fimHmam-ExGene 500 poly cationic polymer and pVAX/fimHwt-ExGene 500 poly cationic polymer. Expression of thefimHgene in both constructs in COS7 cells was confirmed by RT-PCR, SDS-PAGE, and Western blotting. Both of the pVAX/fimHcassettes expressed insertedfimHgenes (mam and wt) in Cos-7 cells. Our results suggest that codon optimization successfully expressed thefimHgene because thefimHgene with mammalian codon usage is compatible with the eukaryotic expression system. Therefore, mammalian codon usage could be appropriate in a pVAX/fimHconstruct as a DNA vaccine.
Antimicrobial susceptibility profiles associated with bacterial meningitis among children: A referral hospital-based study in Iran
Authors: Taghi Haghi-Ashtiani Mohammad , Mamishi Setareh , Shayanfar Nasrin , Mohammadpour Masoud , Yaghmaei Bahareh , Abedini Mina , Farahani Narges , Rezaei Nima
Published: December 2011

Bacterial meningitis continues to be associated with high morbidity and mortality rate worldwide, especially in the pediatric age group. This study was performed to identify the microbial etiologies of meningitis among 31 children, who were admitted in the Emergency Ward of a referral pediatric hospital in Iran. Culture identification showed that Streptococcus pneumoniae (12 subjects), Haemophilus influenzae (11 subjects) were the most common bacteria, followed by Escherichia coli (7 cases) and Neisseria meningitidis (only one case). Antibiotic susceptibility tests revealed that vancomycin had the best effect on S. pneumoniae in comparison with other antibiotics, whereas H. influenzae and E. coli were more susceptible to ceftriaxone, ceftazidime, and ceftizoxime than other antibiotics. In conclusion, despite the advances in antibiotic therapy and vaccine development, bacterial meningitis still is a health problem. S. pneumoniae, H. influenzae, and N. meningitidis are the main sources of bacterial meningitis, but other organisms such as E. coli should also be suspected, when a case is admitted to a referral pediatric hospital.
The effects on immune response of levamisole treatment following infection of U-937 macrophages with Candida albicans
Authors: Kaya Esma , Ozbilge Hatice , Ustundag Mustafa , Torun Yasemin
Published: December 2011

The aim of this study was to investigate the effects on the immune response of levamisole alone and in conjunction withCandida albicansstimulation in human macrophage cell culture by determining the alterations in the levels of cytokine release.Levamisole treatment was performed before, during and after infecting U-937 human macrophage cells withC. albicans. In cell supernatants, interleukin (IL)-1b, IL-12, IL-18, tumour necrosis factor alpha (TNF-α) levels were measured by ELISA.In vitrolevamisole treatment accompanied byC. albicansstimulation significantly increased IL-12, IL-1βand IL-18 production in macrophage cells (p < 0.05). It was observed that when administered beforeC. albicansinfection, levamisole significantly increased IL-12 and IL-1βproduction in macrophage cells (p < 0.05). Another finding was that when applied to macrophage cells simultaneously withC. albicansinfection, or before infection withC. albicans, levamisole suppressed the TNF-βproduction stimulating effect ofC. albicans(p < 0.05).These results indicated that levamisole could be useful in treating patients infected withC. albicansor in protecting individuals under the risk of being infected with this pathogen. There is a need for further experimental and clinical studies on this hypothesis.
Oral Adverse Events Associated with Tyrosine Kinase and Mammalian Target of Rapamycin Inhibitors in Renal Cell Carcinoma: A Structured Literature Review
Authors: Boers-Doets Christine B. , Epstein Joel B. , Raber-Durlacher Judith E. , Ouwerkerk Jan , Logan Richard M. , Brakenhoff Jan A. , Lacouture Mario E. , Gelderblom Hans
Published: December 2011

Learning Objectives

After completing this course, the reader will be able to: Describe the oral manifestations that can appear with TKI/mTORI.Describe the limitations of the current oral assessment tools in assessing these novel presentations and list items needed to assess the presentations properly.

This article is available for continuing medical education credit at CME.TheOncologist.com

Background.

Oral adverse events (OAEs) associated with multitargeted tyrosine kinase inhibitors (TKIs) and mammalian target of rapamycin inhibitors (mTORIs) are underestimated but frequent and novel presentations of mucosal manifestations. Because optimal antitumor activity requires maintaining the optimal dose, it is essential to avoid unintended treatment delays or interruptions.

Methods.

We review the reported prevalence and appearance of OAEs with TKIs and mTORIs and the current oral assessment tools commonly used in clinical trials. We discuss the correlations between OAEs and hand–foot skin reaction (HFSR) and rash.

Results.

The reported prevalence of oral mucositis/stomatitis of any grade is 4% for pazopanib, 28% for sorafenib, 38% for sunitinib, 41% for temsirolimus, and 44% for everolimus. Oral lesions associated with these agents have been reported to more closely resemble aphthous stomatitis than OM caused by conventional agents. In addition, these agents may result in symptoms such as oral mucosal pain, dysgeusia, and dysphagia, in the absence of clinical lesions. Because of these factors, OAEs secondary to targeted agents may be underreported. In addition, a correlation between OAEs and HFSR was identified.

Conclusions.

OAEs caused by TKIs and mTORIs may represent dose-limiting toxicities, especially considering the fact that even low grades of OAEs may be troubling to the patient. We discuss how these novel AEs can be assessed because current mucositis assessment tools have limitations. Prospective studies investigating the pathogenesis, risk factors, and management of OAEs are needed in order to minimize the impact on patient's health-related quality of life.
Optimization of Extraction of D-pinitol and Phenolics from Cultivated and Wild Types of Carob Pods Using Response Surface Methodology
Authors: Turhan Irfan
Published: December 2011

The carob pod, an important fruit of the food industry in the production of concentrated syrup, is rich in potentially health-promoting phenolic compounds and sugars. Moreover, one of the important bioactive components of carob is D-pinitol, and it has some beneficial effects on human metabolism. The optimum conditions for extraction of D-pinitol and total phenolic compounds (TPC) from cultivated and wild types of carob pods were determined using response surface methodology (RSM). The Box-Behnken Design (BBD) was used to investigate the effects of three independent variables; extraction temperature (ºC), dilution rate (w/v), and extraction time (h) on the response, D-pinitol and the total phenolic compounds. The optimum extraction conditions obtained using the response optimizer were an extraction temperature of 80ºC, a dilution rate of 1:4, and an extraction time of 2h. Under the mentioned above conditions, the maximum D-pinitol and TPC concentrations were 9.67 g/L and 5916.55 mg/L in wild carob pod extract, respectively.
Organic carbon stock in some forest soils in Serbia
Authors: Kadovic Ratko , Belanovic Snezana , Knezevic Milan , Danilovic Milorad , Kosanin Olivera , Beloica Jelena
Published: December 2011

The content of organic carbon (C) was researched in topsoil layers (0-20 cm)
in the most represented soils of forest ecosystems in central Serbia: eutric
ranker, eutric cambisol and dystric cambisol. The soils were sampled during
2003, 2004 and 2010. Laboratory analyses included the soil physical and
chemical properties necessary for the quantification of the soil organic
carbon in organic and mineral layers. Mean values of the soil organic carbon
(SOC) stores in organic horizons of the study soils varied between: 1.01?0.4
kg(C).m-2 (dystric cambisol), 0.90?0.41 kg(C).m-2 (eutric ranker) and
0.94?0.36 kg(C).m-2 (eutric cambisol). Average values of organic carbon in
mineral layers (0-20 cm) ranged between: 3.83?1.70 kg(C).m-2 (dystric
cambisol), 6.26?3.41 kg(C).m-2 (eutric ranker) and 4.36?1.91 kg(C).m-2
(eutric cambisol). The average value of total organic carbon stock in the
study soils (both organic and mineral layers) was 5.77 kg(C).m-2. This paper
addresses the methodological aspects of regional estimation of soil organic
carbon content as the potential to be applied in the National Forest
Inventory Program.
Hepatic differentiation from human mesenchymal stem cells on a novel nanofiber scaffold
Authors: Ghaedi Mahboobe , Soleimani Masoud , Shabani Iman , Duan Yuyou , Lotfi Abbas
Published: December 2011

The emerging fields of tissue engineering and biomaterials have begun to provide potential treatment options for liver failure. The goal of the present study is to investigate the ability of a poly L-lactic acid (PLLA) nanofiber scaffold to support and enhance hepatic differentiation of human bone marrow-derived mesenchymal stem cells (hMSCs). A scaffold composed of poly L-lactic acid and collagen was fabricated by the electrospinning technique. After characterizing isolated hMSCs, they were seeded onto PLLA nanofiber scaffolds and induced to differentiate into a hepatocyte lineage. The mRNA levels and protein expression of several important hepatic genes were determined using RT-PCR, immunocytochemistry and ELISA. Flow cytometry revealed that the isolated bone marrow-derived stem cells were positive for hMSC-specific markers CD73, CD44, CD105 and CD166 and negative for hematopoietic markers CD34 and CD45. The differentiation of these stem cells into adipocytes and osteoblasts demonstrated their multipotency. Scanning electron microscopy showed adherence of cells in the nanofiber scaffold during differentiation towards hepatocytes. Our results showed that expression levels of liver-specific markers such as albumin, α-fetoprotein, and cytokeratins 8 and 18 were higher in differentiated cells on the nanofibers than when cultured on plates. Importantly, liver functioning serum proteins, albumin and α-1 antitrypsin were secreted into the culture medium at higher levels by the differentiated cells on the nanofibers than on the plates, demonstrating that our nanofibrous scaffolds promoted and enhanced hepatic differentiation under our culture conditions. Our results show that the engineered PLLA nanofibrous scaffold is a conducive matrix for the differentiation of MSCs into functional hepatocyte-like cells. This represents the first step for the use of this nanofibrous scaffold for culture and differentiation of stem cells that may be employed for tissue engineering and cell-based therapy applications.
$\bar \lambda $-statistically convergent double sequences in probabilistic normed spaces
Authors: Savaş E. , Mohiuddine S.
Published: December 2011

The purpose of this paper is to introduce and study the concepts of double $\bar \lambda $-statistically convergent and double $\bar \lambda $-statistically Cauchy sequences in probabilistic normed space.
Connectedness of the sets of weak efficient solutions for generalized vector equilibrium problems
Authors: Liu Qing-You , Long Xian-Jun , Huang Nan-jing
Published: December 2011

In this paper, a generalized vector equilibrium problem is introduced and studied. A scalar characterization of weak efficient solutions for the generalized vector equilibrium problem is obtained. By using the scalarization result, the existence of the weak efficient solutions and the connectedness of the set of weak efficient solutions for the generalized vector equilibrium problem are proved in locally convex spaces.
Transcranial magnetic resonance imaging–guided focused ultrasound: noninvasive central lateral thalamotomy for chronic neuropathic pain
Authors: Jeanmonod Daniel , Werner Beat , Morel Anne , Michels Lars , Zadicario Eyal , Schiff Gilat , Martin Ernst
Published: December 2011

Object

Recent technological developments open the field of therapeutic application of focused ultrasound to the brain through the intact cranium. The goal of this study was to apply the new transcranial magnetic resonance imaging–guided focused ultrasound (tcMRgFUS) technology to perform noninvasive central lateral thalamotomies (CLTs) as a treatment for chronic neuropathic pain.

Methods

In 12 patients suffering from chronic therapy-resistant neuropathic pain, tcMRgFUS CLT was proposed. In 11 patients, precisely localized thermal ablations of 3–4 mm in diameter were produced in the posterior part of the central lateral thalamic nucleus at peak temperatures between 51°C and 64°C with the aid of real-time patient monitoring and MR imaging and MR thermometry guidance. The treated neuropathic pain syndromes had peripheral (5 patients) or central (6 patients) origins and covered all body parts (face, arm, leg, trunk, and hemibody).

Results

Patients experienced mean pain relief of 49% at the 3-month follow-up (9 patients) and 57% at the 1-year follow-up (8 patients). Mean improvement according to the visual analog scale amounted to 42% at 3 months and 41% at 1 year. Six patients experienced immediate and persisting somatosensory improvements. Somatosensory and vestibular clinical manifestations were always observed during sonication time because of ultrasound-based neuronal activation and/or initial therapeutic effects. Quantitative electroencephalography (EEG) showed a significant reduction in EEG spectral overactivities. Thermal ablation sites showed sharply delineated ellipsoidal thermolesions surrounded by short-lived vasogenic edema. Lesion reconstructions (18 lesions in 9 patients) demonstrated targeting precision within a millimeter for all 3 coordinates. There was 1 complication, a bleed in the target with ischemia in the motor thalamus, which led to the introduction of 2 safety measures, that is, the detection of a potential cavitation by a cavitation detector and the maintenance of sonication temperatures below 60°C.

Conclusions

The authors assert that tcMRgFUS represents a noninvasive, precise, and radiation-free neurosurgical technique for the treatment of neuropathic pain. The procedure avoids mechanical brain tissue shift and eliminates the risk of infection. The possibility of applying sonication thermal spots free from trajectory restrictions should allow one to optimize target coverage. The real-time continuous MR imaging and MR thermometry monitoring of targeting accuracy and thermal effects are major factors in optimizing precision, safety, and efficacy in an outpatient context.