Genetic and epigenetic changes in primary metastatic and nonmetastatic colorectal cancer

Colorectal cancer (CRC) develops as multistep process, which involves genetic and epigenetic alterations. K-Ras, p53 and B-Raf mutations and RASSF1A, E-Cadherin and p16INK4A promoter methylation were investigated in 202 CRCs with and without lymph node and/or liver metastasis, to assess whether gene abnormalities are related to a metastogenic phenotype. K-Ras, B-Raf and p53 mutations were detected in 27, 3 and 32% of the cases, with K-Ras mutations significantly associated with metastatic tumour (P=0.019). RASSF1A, E-Cadherin and p16INK4A methylation was documented in 20, 44 and 33% of the cases with p16INK4A significantly associated with metastatic tumours (P=0.001). Overall, out of 202 tumours, 34 (17%) did not show any molecular change, 125 (62%) had one or two and 43 (21%) three or more. Primary but yet metastatic CRCs were prevalent in the latter group (P=0.023) where the most frequent combination was one genetic (K-Ras in particular) and two epigenetic alterations. In conclusion, this analysis provided to detect some molecular differences between primary metastatic and nonmetastatic CRCs, with K-Ras and p16INK4A statistically altered in metastatic tumours; particular gene combinations, such as coincidental K-Ras mutation with two methylated genes are associated to a metastogenic phenotype

Selective staining and eradication of cancer cells by protein-carrying DARPin-functionalized liposomes

Since their discovery, liposomes have been widely employed in biomedical research. These nano-size spherical vesicles consisting one or few phospholipid bilayers surrounding an aqueous core are capable of carrying a wide variety of bioactive compounds, including drugs, peptides, nucleic acids, proteins and others. Despite considerable success achieved in synthesis of liposome constructs containing bioactive compounds, preparation of ligand-targeted liposomes comprising large quantities of encapsulated proteins that are capable of affecting pathological cells still remains a big challenge. Here we described a novel method for preparation of small (80\u201390 nm in diameter) unilamellar liposomes containing very large quantities (thousands of protein molecules per liposome) of heme-containing cytochrome c, highly fluorescent mCherry and highly toxic PE40 (Pseudomonas aeruginosa Exotoxin A domain). Efficient encapsulation of the proteins was achieved through electrostatic interaction between positively charged proteins (at pH lower than pI) and negatively charged liposome membrane. The proteoliposomes containing large quantities of mCherry or PE40 and functionalized with designed ankyrin repeat protein (DARPin)_9-29, which targets human epidermal growth factor receptor 2 (HER2) were shown to specifically stain and kill in sub-nanomolar concentrations HER2-positive cells, overexpressing HER2, respectively. Specific staining and eradication of the receptor-positive cells demonstrated here makes the DARPin-functionalized liposomes carrying large quantities of fluorescent and/or toxic proteins a promising candidate for tumor detection and therapy

Combinatorial CRISPR-Cas9 screens for de novo mapping of genetic interactions.

We developed a systematic approach to map human genetic networks by combinatorial CRISPR-Cas9 perturbations coupled to robust analysis of growth kinetics. We targeted all pairs of 73 cancer genes with dual guide RNAs in three cell lines, comprising 141,912 tests of interaction. Numerous therapeutically relevant interactions were identified, and these patterns replicated with combinatorial drugs at 75% precision. From these results, we anticipate that cellular context will be critical to synthetic-lethal therapies

Скрининг латентной туберкулезной инфекции с применением аллергена туберкулезного рекомбинантного

In recent years the significant deterioration of health has been observed in schoolchildren. As per official data only 20% of children graduating from school are considered to be completely healthy, and 60% suffer from chronic diseases, i.e. belong to health groups III, IV and V, among which there are many children belonging to tuberculosis risk groups and suffering from undetected latent tuberculous infection. The efficiency of the new technique aimed to detect active tuberculous infection in children and adolescents of health groups III, IV and V has been evaluated. Screening proved the reduction in the number of patients in need of TB doctor advising by 8.4 fold compared to traditional mass diagnostics with tuberculin. Hyperergic reactions to diaskintest are 6 fold less common compared to Mantoux test. And 99.2% of children referred to TB doctor were registered to the dispensary follow-up versus 23.4% of children referred to TB doctor as per mass tuberculin testing results. (p < 0.001). Thus the validity of referral to TB doctor increased 4.2 fold. The detection rate of active forms of tuberculosis made 0.4 per 1000 examinations versus 0.1 in case of mass screening with tuberculin testing.В последние годы наблюдается существенное ухудшение состояния здоровья школьников. По официальным данным, лишь 20% детей, заканчивающих школу, считаются полностью здоровыми, а 60% имеют хронические заболеваниями, т. е. III, IV или V группу здоровья, среди которых велика доля детей, относящихся к группам риска по туберкулезу и имеющих невыявленную латентную туберкулезную инфекцию. Проведена оценка эффективности нового метода выявления активной туберкулезной инфекции у детей и подростков III, IV и V групп здоровья. Скрининговое исследование показало уменьшение числа пациентов, нуждающихся в консультации фтизиатра, в 8,4 раза по сравнению с традиционной массовой туберкулинодиагностикой. Гиперергические реакции на пробу с диаскинтестом выявляются в 6 раз реже по сравнению с пробой Манту. При этом на диспансерный учет у врача-фтизиатра взято 99,2% детей от направленных на консультацию против 23,4% детей, направленных по результатам массовой туберкулинодиагностики (p < 0,001). Таким образом, в 4,2 раза увеличилась обоснованность направления ребенка к врачу-фтизиатру. Выявляемость активных форм туберкулеза новым методом составила 0,4 на 1 000 обследованных против 0,1 при проведении массовой традиционной туберкулинодиагностики

Dbf4-dependent kinase (DDK)-mediated proteolysis of CENP-A prevents mislocalization of CENP-A in Saccharomyces cerevisiae

The evolutionarily conserved centromeric histone H3 variant (Cse4 in budding yeast, CENP-A in humans) is essential for faithful chromosome segregation. Mislocalization of CENP-A to non-centromeric chromatin contributes to chromosomal instability (CIN) in yeast, fly, and human cells and CENP-A is highly expressed and mislocalized in cancers. Defining mechanisms that prevent mislocalization of CENP-A is an area of active investigation. Ubiquitin-mediated proteolysis of overexpressed Cse4 (GALCSE4)byE3 ubiquitin ligases such as Psh1 prevents mislocalization of Cse4, and psh1D strains display synthetic dosage lethality (SDL) with GALCSE4. We previously performed a genome-wide screen and identified five alleles of CDC7 and DBF4 that encode the Dbf4-dependent kinase (DDK) complex, which regulates DNA replication initiation, among the top twelve hits that displayed SDL with GALCSE4. We determined that cdc7-7 strains exhibit defects in ubiquitin-mediated proteolysis of Cse4 and show mislocalization of Cse4. Mutation of MCM5 (mcm5-bob1) bypasses the requirement of Cdc7 for replication initiation and rescues replication defects in a cdc7-7 strain. We determined that mcm5-bob1 does not rescue the SDL and defects in proteolysis of GALCSE4 in a cdc7-7 strain, suggesting a DNA replication-independent role for Cdc7 in Cse4 proteolysis. The SDL phenotype, defects in ubiquitin-mediated proteolysis, and the mislocalization pattern of Cse4 in a cdc7-7 psh1D strain were similar to that of cdc7-7 and psh1D strains, suggesting that Cdc7 regulates Cse4 in a pathway that overlaps with Psh1. Our results define a DNA replication initiation-independent role of DDK as a regulator of Psh1-mediated proteolysis of Cse4 to prevent mislocalization of Cse4.Fil: Eisenstatt, Jessica R.. National Institutes of Health; Estados UnidosFil: Boeckmann, Lars. National Institutes of Health; Estados UnidosFil: Au, Wei Chun. National Institutes of Health; Estados UnidosFil: Garcia, Valerie. National Institutes of Health; Estados UnidosFil: Bursch, Levi. National Institutes of Health; Estados UnidosFil: Ocampo, Josefina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina. National Instituto of Child Health & Human Development; Estados UnidosFil: Costanzo, Michael. National Institutes of Health; Estados Unidos. University of Toronto; CanadáFil: Weinreich, Michael. Van Andel Research Institute; Estados UnidosFil: Sclafani, Robert A.. University of Colorado; Estados UnidosFil: Baryshnikova, Anastasia. University of Princeton; Estados UnidosFil: Myers, Chad L.. University of Minnesota; Estados UnidosFil: Boone, Charles. University of Toronto; Canadá. National Institutes of Health; Estados UnidosFil: Clark, David J.. National Institutes of Health; Estados UnidosFil: Baker, Richard. University of Massachusetts; Estados UnidosFil: Basrai, Munira A.. National Institutes of Health; Estados Unido

The rapeutic effects of gestational contraception

The purpose of the study is to study the clinical efficacy of the etonogestrel subcutaneous implant.Цель исследования – изучение клинической эффективности подкожного импланта этоногестрела

Скрининг детей и подростков на туберкулезную инфекцию в России – прошлое, настоящее, будущее

The article presents a brief overview of the establishment and development of screening system in the pediatric population for tuberculosis and latent tuberculosis infection - from Koch alt-tuberculin to modern recombinant tuberculosis allergens for intradermal administration and in vitro laboratory tests. It gives extensive data on clinical research and the introduction into practical public health of the Russian Federation of the test with recombinant tuberculosis allergen as a screening method for tuberculosis in children from 8 to 14 years old inclusively.Представлен краткий обзор становления и развития системы скрининга детского населения на туберкулез и на латентную туберкулезную инфекцию – от альт-туберкулина Коха до современных рекомбинантных аллергенов туберкулезных для внутрикожного введения и лабораторных тестов in vitro. Приведены обширные данные о клиническом исследовании и внедрении в практическое здравоохранение РФ пробы с аллергеном туберкулезным рекомбинантным в качестве скрининга на туберкулез у детей с 8 до 14 лет включительно

Assessment of the prevalence of somatic pathology in women in the gestational period

The purpose of the study is a comparative analysis of the structure of extragenital pathology in primiparous women of different age groups.Цель исследования – сравнительный анализ структуры экстрагенитальной патологии у первородящих женщин разных возрастных групп

Milestones in the Observations of Cosmic Magnetic Fields

Magnetic fields are observed everywhere in the universe. In this review, we concentrate on the observational aspects of the magnetic fields of Galactic and extragalactic objects. Readers can follow the milestones in the observations of cosmic magnetic fields obtained from the most important tracers of magnetic fields, namely, the star-light polarization, the Zeeman effect, the rotation measures (RMs, hereafter) of extragalactic radio sources, the pulsar RMs, radio polarization observations, as well as the newly implemented sub-mm and mm polarization capabilities. (Another long paragraph is omitted due to the limited space here)Comment: Invited Review (ChJA&A); 32 pages. Sorry if your significant contributions in this area were not mentioned. Published pdf & ps files (with high quality figures) now availble at http://www.chjaa.org/2002_2_4.ht