Neoglycoprotein binding to colorectal tumour cells: Comparison between primary and secondary lesions

Summary Biotinylated neoglycoproteins are useful to determine the expression of sugar receptors (lectins) histochemically in routinely processed tissue sections. Assessment of the presence of distinct receptor classes with specificity to-galactosides and to- or-N-acetylgalactosamine, selected on the basis of their potential relevance for recognition processes within the metastatic cascade in murine model systems, was performed for a common human tumour type, colorectal cancer. The four different types of neoglycoproteins, derived from covalent attachment of commercially available derivatives of-N-acetylgalactosamine, differed only quantitatively in their capacity to detect specific binding on cultured cells and tissue sections, thus posing no major restriction on the choice of synthetic process for histochemical efficiency of the product. Glycocytological application revealed specific probe binding and a regulation of level of receptor expression for a human colon carcinoma cell line primarily forN-acetylgalactosamine-specific receptors upon retinoic acid-induced differentiation. Monitoring of sections of the 12 cases of primary and secondary colorectal lesions invariably disclosed the presence of the respective receptors, the extent of cell labelling in primary tumours and metastases being similar. Establishment of metastases, even in different target organs, is apparently not followed by a major phenotypic variation in this feature.     

Resistance mechanisms to plant viruses: an overview

To obtain virus-resistant host plants, a range of operational strategies can be followed nowadays. While for decades plant breeders have been able to introduce natural resistance genes in susceptible genotypes without knowing precisely what these resistance traits were, currently a growing number of (mostly) dominant resistance genes have been cloned and analyzed. This has led not only to a better understanding of the plant's natural defence systems, but also opened the way to use these genes beyond species borders. Besides using natural resistance traits, also several novel, "engineered" forms of virus resistance have been developed over the past 15 years. The first successes were obtained embarking from the principle of pathogen-derived resistance (PDR) by transforming host plants with viral genes or sequences with the purpose to block a specific step during virus multiplication in the plant. As an unforeseen spin-off of these investments, the phenomenon of post-translational gene silencing (PTGS) was discovered, which to date is by far the most successful way to engineer resistance. It is generally believed that PTGS reflects a natural defence system of the plant, and part of the hypothesized components required for PTGS have been identified. As counteracting strategy, and confirming PTGS to be a natural phenomenon, a considerable number of viruses have acquired gene functions by which they can suppress PTGS. In addition to PDR and PTGS, further strategies for engineered virus resistance have been explored, including the use of pokeweed antiviral protein (PAP), 2',5'-oligoadenylate synthetase and "plantibodies". This paper will give a brief overview of the major strategies that have become operational during the past 10 years.     

Alteration of the LRP1B gene region is associated with high grade of urothelial cancer

We have delineated regions of interest at chromosome 2q21.2, 2q36.3, and 2q37.1 by deletion mapping of 114 urothelial cancers (UC). Altogether, 17%, 18%, and 63% of the G1, G2, and G3 tumors displayed loss of heterozygosity at chromosome 2q, respectively, The region at 2q21.2 was narrowed down to the LRP1B gene (NT_005129.6). Hemi- and homozygous deletion at the LRP1B gene region was seen in 31 of 114 UCs. Only 8% of the UCs with G1 and none with G2 tumors showed loss of heterozygosity at the LRP1B gene, whereas 49% of the G3 UCs had allelic loss at this region. RT-PCR analysis of the LRP1B gene showed the lack of expression of several exons in 2 of 9 cases analyzed. Our analysis suggests that the LRP1B gene is a candidate tumor suppressor gene in UCs.     

Molecular cloning and sequence analysis of the 3′-terminal region of iris severe mosaic virus RNA

Summary Sequence analysis of iris severe mosaic potyvirus genomic RNA revealed an unusual E/G cleavage site between the deduced large nuclear-inclusion protein and coat protein sequences. The latter showed an N-terminus of only 15 amino acids. The 3 non-translated region of the viral RNA was 340 nucleotides long.     

Cyberbullying Perpetration and Victimization Among Middle-School Students

Objectives. We examined correlations between gender, race, sexual identity, and technology use, and patterns of cyberbullying experiences and behaviors among middle-school students.Methods. We collected a probability sample of 1285 students alongside the 2012 Youth Risk Behavior Survey in Los Angeles Unified School District middle schools. We used logistic regressions to assess the correlates of being a cyberbully perpetrator, victim, and perpetrator–victim (i.e., bidirectional cyberbullying behavior).Results. In this sample, 6.6% reported being a cyberbully victim, 5.0% reported being a perpetrator, and 4.3% reported being a perpetrator–victim. Cyberbullying behavior frequently occurred on Facebook or via text messaging. Cyberbully perpetrators, victims, and perpetrators–victims all were more likely to report using the Internet for at least 3 hours per day. Sexual-minority students and students who texted at least 50 times per day were more likely to report cyberbullying victimization. Girls were more likely to report being perpetrators–victims.Conclusions. Cyberbullying interventions should account for gender and sexual identity, as well as the possible benefits of educational interventions for intensive Internet users and frequent texters.Cyberbullying is the “willful and repeated harm inflicted [on another] through the use of computers, cell phones, or other electronic devices.”1^(p5) Among 6th- through 10th-grade students nationally, 4% reported being cyberbullying perpetrators, 5% reported being a victim of cyberbullying, and 5% reported being perpetrators–victims (meaning that they have both perpetrated and been victimized by cyberbullying) during the previous 2 months.2 Among middle-school students in the southeastern and northwestern United States, a nonprobability sample of 3767 students by Kowalski and Limber3 found that 11% reported being a victim of cyberbullying, 4% were cyberbullying perpetrators, and 7% were perpetrators–victims during the previous 2 months. Cyberbullying may be more insidious than traditional bullying, because cyberbullying can quickly reach wide audiences (e.g., e-mails sent to an entire school), can be perpetrated anonymously, and is not bound to in-person interactions.4,5 Although cyberbullying has garnered widespread media attention, to our knowledge, no previous study has explored correlates of cyberbullying with a representative probability sample in an urban middle-school sample.Cyberbullying is associated with a host of health and behavioral health consequences. Research has suggested that cyberbullying may have a greater effect on depression and suicidal ideation than traditional offline bullying.6 Both perpetration and victimization are associated with mental health consequences, including lower self-esteem,7 recent depressive symptoms,5,8,9 and suicidal ideation.5,8,10 Cyberbullying perpetrators are more likely to have problems with their behavior, peer relationships, and emotions, and are less likely to be prosocial than their peers who are neither cyberbullying perpetrators nor victims of cyberbullying.11 Specifically, female cyberbullying perpetrators express greater anxiety and depression than their female peers who are not cyberbullying perpetrators.12 Cyberbullying victimization is also strongly associated with substance use, violent behavior, and risky sexual behavior among high-school students.10Cyberbullying disproportionately affects youths who are already vulnerable to mental health and behavioral health disparities, including members of sexual minorities (i.e., gay, lesbian, bisexual), girls, and racial and ethnic minorities. More than half of sexual-minority middle- and high-school students nationally report being a cyberbully victim during the previous year, with almost one fifth reporting often or frequent victimization.13 Female students are significantly more likely to be cyberbully perpetrators–victims than their male peers.9,14 Studies have reported that male students are significantly more likely to be cyberbullying perpetrators2,15 and significantly less likely to be cyberbully victims.2,6,9 Middle-school boys are more likely to cyberbully others because of their race, sexual identity, or both.16 African American students are more likely to be cyberbully perpetrators and Hispanic students are more likely to be cyberbully perpetrators–victims than their White peers.2Cyberbullying may occur across a variety of technology platforms, and the specific qualities of each platform may affect how cyberbullying is perpetrated and experienced. Earlier studies suggested that instant messaging, chat rooms, and message boards were the most common mediums for middle-school students who experienced cyberbullying.3,17 More recently, youths have migrated to social media platforms such as Facebook,18–20 necessitating an examination of cyberbullying across more contemporary and emerging platforms for youth interaction.More frequent use of technology has also been associated with cyberbullying. Students who use the Internet for at least 3 hours per day and those who use instant messaging and Web cams are significantly more likely to have been cyberbully victims at least 7 times during the previous year.17 Frequent Internet users are significantly more likely to be cyberbully perpetrators, victims, and perpetrators–victims.14Research has shown that even though cyberbullying takes place in a virtual space, most cyberbullying perpetrators know their victims and vice versa. Moreover, 73% of victims reported being “pretty sure” or “totally sure” about the identity of their cyberbully, with 51% of cyberbullying perpetrators identified as a classmate, 43% as someone who they only knew online, and 20% as an in-person, nonclassmate relation.17 Cyberbullying perpetrators of middle-school victims were most often a classmate or a stranger; cyberbullying perpetrators most often reported that they cyberbullied classmates, friends, and strangers.3This study expanded previous work with high-school samples2 and nonprobability samples of middle-school students3,17 to explore the role of sexual identity, gender, race, and technology use patterns in a random sample of urban middle-school students in Los Angeles County, California. The study focused on demographic characteristics including sexual identity, patterns of technology use (i.e., frequency of Internet use, texting, parental rules regarding Internet use), and platforms on which middle-school students experience cyberbullying (i.e., Facebook, Twitter, e-mail, text messaging) to inform interventions to disrupt this maladaptive behavior and help protect early adolescents from the consequences of cyberbullying.     

Effect of ozone on periodontopathogenic species—an in vitro study

The in vitro study was aimed to determine the effect of ozone on periodontopathogenic microorganisms. Ozone was generated for 6 s–2 × 24 s (corresponding to 0.56 mg–2 × 2.24 mg of ozone) against 23 mainly anaerobic periodontopathogenic species. Agar diffusion test was used as a screening method. Then, the killing activity was tested in a serum-free environment and with 25% v/v inactivated serum. Further, the effect of ozone on bactericidal activity of native serum was analyzed against Fusobacterium nucleatum, Porphyromonas gingivalis, and Aggregatibacter actinomycetemcomitans. Agar diffusion test showed a high efficacy of ozone against microorganisms, especially against Porphyromonas gingivalis. This result was confirmed by the killing tests; most of the strains in a concentration of 10⁵ were completely eliminated after twofold 18-s application of ozone. Only four of the six potentially “superinfecting” species (Staphylococcus aureus, Enterococcus faecalis, Enterobacter cloacae, Candida albicans) survived in part. Addition of heat-inactivated serum reduced the killing rate of ozone by 78% after 6-s and by 47% after twofold 18-s exposures; no strain was completely eradicated after any application of ozone. The bactericidal effect of native serum was enhanced after application of ozone; no effect was visible on the included A. actinomycetemcomitans strain which was found to be completely resistant to the bactericidal action of serum. In conclusion, (a) ozone has a strong antibacterial activity against putative periodontopathogenic microorganisms, and (b) the bactericidal effect is reduced in the presence of serum. Ozone may have potential as an adjunctive application to mechanical treatment in periodontitis patients.     

The potential association between gingival crevicular fluid inflammatory mediators and adverse pregnancy outcomes: a systematic review

Objectives

The association between periodontal disease and adverse pregnancy outcomes (APO), primarily preterm birth (PTB), is still controversially discussed in the literature. Therefore, the aim of the present systematic review was to analyze the existing literature on the potential association between inflammatory mediators detected in gingival crevicular fluid (GCF) and APO.

Materials and methods

MEDLINE (PubMed) and EMBASE databases were searched for entries up to April 2012 and studies were selected by two independent reviewers.

Results

The majority of the eight studies included confirmed a positive association between GCF mediators, such as interleukin-1β, prostaglandin E₂, and tumor necrosis factor-alpha, and APO. Due to the heterogeneity and variability of the available studies, no meta-analysis could be performed.

Conclusions

A positive association between GCF inflammatory mediator levels and APO/PTB might be present but the results need to be considered with great caution because of the heterogeneity and variability among the studies. Further studies with an adequate number of patients allowing for an appropriate analysis are warranted to definitely confirm this association.

Clinical relevance

The present findings suggest that an association between GCF inflammatory mediator levels and APO might exist.     

Extracellular vesicles or free circulating DNA: where to search for BRAF and cKIT mutations?

Clinical evidence in oncology argues for the advantages of performing molecular analysis of blood biomarkers to provide information about systemic changes and tumor heterogeneity.Whereas the diagnostic value of cell-free circulating DNA (fcDNA) has successfully been demonstrated in several studies, DNA enclosed in extracellular vesicles (EV) has only recently been described, and its potential diagnostic value is unclear. We established a protocol for separation of EV and fc fractions and tested for presence of mutant BRAFV600E mediating resistance to Vemurafenib and cKITD816V mediating resistance to Imatinib in blood of patients with melanoma and mastocytosis. Our results show that EV contain significantly higher amounts of total DNA as compared to the fc fraction. However, about ten-fold higher copy numbers of the wild type and mutant BRAF and cKIT were detected in the fcDNA fraction supporting its diagnostic value and pointing to differences in fc and EV DNA content.