A narnavirus-like element from the trypanosomatid protozoan parasite Leptomonas seymouri

Genome sequences were determined for a novel RNA virus, Leptomonas seymouri Narna-like virus 1 (LepseyNLV1). A 2.9-kb segment encodes an RNA-dependent RNA polymerase (RdRp), while a smaller 1.5-kb segment showed no database search matches. This is the first report of bisegmented Narnaviridae from insect trypanosomatids

A Narnavirus in the trypanosomatid protist plant pathogen Phytomonas serpens

We describe here a new RNA virus (PserNV1) from the plant protist parasite Phytomonas serpens (family Trypanosomatidae, Kinetoplastida, supergroup Excavata). The properties of PserNV1 permit assignment to the genus Narnavirus (Narnaviridae), the first reported from a host other than fungi or oomycetes

A novel bunyavirus-like virus of trypanosomatid protist parasites

We report here the sequences for all three segments of a novel RNA virus (LepmorLBV1) from the insect trypanosomatid parasite Leptomonas moramango. This virus belongs to a newly discovered group of bunyavirus-like elements termed Leishbunyaviruses (LBV), the first discovered from protists related to arboviruses infecting humans

Leishmania aethiopica field isolates bearing an endosymbiontic dsRNA virus induce pro-inflammatory cytokine response.

BACKGROUND: Infection with Leishmania parasites causes mainly cutaneous lesions at the site of the sand fly bite. Inflammatory metastatic forms have been reported with Leishmania species such as L. braziliensis, guyanensis and aethiopica. Little is known about the factors underlying such exacerbated clinical presentations. Leishmania RNA virus (LRV) is mainly found within South American Leishmania braziliensis and guyanensis. In a mouse model of L. guyanensis infection, its presence is responsible for an hyper-inflammatory response driven by the recognition of the viral dsRNA genome by the host Toll-like Receptor 3 leading to an exacerbation of the disease. In one instance, LRV was reported outside of South America, namely in the L. major ASKH strain from Turkmenistan, suggesting that LRV appeared before the divergence of Leishmania subgenera. LRV presence inside Leishmania parasites could be one of the factors implicated in disease severity, providing rationale for LRV screening in L. aethiopica. METHODOLOGY/PRINCIPAL FINDINGS: A new LRV member was identified in four L. aethiopica strains (LRV-Lae). Three LRV-Lae genomes were sequenced and compared to L. guyanensis LRV1 and L. major LRV2. LRV-Lae more closely resembled LRV2. Despite their similar genomic organization, a notable difference was observed in the region where the capsid protein and viral polymerase open reading frames overlap, with a unique -1 situation in LRV-Lae. In vitro infection of murine macrophages showed that LRV-Lae induced a TLR3-dependent inflammatory response as previously observed for LRV1. CONCLUSIONS/SIGNIFICANCE: In this study, we report the presence of an immunogenic dsRNA virus in L. aethiopica human isolates. This is the first observation of LRV in Africa, and together with the unique description of LRV2 in Turkmenistan, it confirmed that LRV was present before the divergence of the L. (Leishmania) and (Viannia) subgenera. The potential implication of LRV-Lae on disease severity due to L. aethiopica infections is discussed

Tilting the balance between RNA interference and replication eradicates Leishmania RNA virus 1 and mitigates the inflammatory response.

Many Leishmania (Viannia) parasites harbor the double-stranded RNA virus Leishmania RNA virus 1 (LRV1), which has been associated with increased disease severity in animal models and humans and with drug treatment failures in humans. Remarkably, LRV1 survives in the presence of an active RNAi pathway, which in many organisms controls RNA viruses. We found significant levels (0.4 to 2.5%) of small RNAs derived from LRV1 in both Leishmania braziliensis and Leishmania guyanensis, mapping across both strands and with properties consistent with Dicer-mediated cleavage of the dsRNA genome. LRV1 lacks cis- or trans-acting RNAi inhibitory activities, suggesting that virus retention must be maintained by a balance between RNAi activity and LRV1 replication. To tilt this balance toward elimination, we targeted LRV1 using long-hairpin/stem-loop constructs similar to those effective against chromosomal genes. LRV1 was completely eliminated, at high efficiency, accompanied by a massive overproduction of LRV1-specific siRNAs, representing as much as 87% of the total. For both L. braziliensis and L. guyanensis, RNAi-derived LRV1-negative lines were no longer able to induce a Toll-like receptor 3-dependent hyperinflammatory cytokine response in infected macrophages. We demonstrate in vitro a role for LRV1 in virulence of L. braziliensis, the Leishmania species responsible for the vast majority of mucocutaneous leishmaniasis cases. These findings establish a targeted method for elimination of LRV1, and potentially of other Leishmania viruses, which will facilitate mechanistic dissection of the role of LRV1-mediated virulence. Moreover, our data establish a third paradigm for RNAi-viral relationships in evolution: one of balance rather than elimination

Inhibitor of growth protein 3 epigenetically silences endogenous retroviral elements and prevents innate immune activation

Endogenous retroviruses (ERVs) are subject to transcriptional repression in adult tissues, in part to prevent autoimmune responses. However, little is known about the epigenetic silencing of ERV expression. Here, we describe a new role for inhibitor of growth family member 3 (ING3), to add to an emerging group of ERV transcriptional regulators. Our results show that ING3 binds to several ERV promoters (for instance MER21C) and establishes an EZH2-mediated H3K27 trimethylation modification. Loss of ING3 leads to decreases of H3K27 trimethylation enrichment at ERVs, induction of MDA5-MAVS-interferon signaling, and functional inhibition of several virus infections. These data demonstrate an important new function of ING3 in ERV silencing and contributing to innate immune regulation in somatic cells

Leishmania aethiopica field isolates bearing an endosymbiontic dsRNA virus induce pro-inflammatory cytokine response

Infection with Leishmania parasites causes mainly cutaneous lesions at the site of the sand fly bite. Inflammatory metastatic forms have been reported with Leishmania species such as L. braziliensis, guyanensis and aethiopica. Little is known about the factors underlying such exacerbated clinical presentations. Leishmania RNA virus (LRV) is mainly found within South American Leishmania braziliensis and guyanensis. In a mouse model of L. guyanensis infection, its presence is responsible for an hyper-inflammatory response driven by the recognition of the viral dsRNA genome by the host Toll-like Receptor 3 leading to an exacerbation of the disease. In one instance, LRV was reported outside of South America, namely in the L. major ASKH strain from Turkmenistan, suggesting that LRV appeared before the divergence of Leishmania subgenera. LRV presence inside Leishmania parasites could be one of the factors implicated in disease severity, providing rationale for LRV screening in L. aethiopica.A new LRV member was identified in four L. aethiopica strains (LRV-Lae). Three LRV-Lae genomes were sequenced and compared to L. guyanensis LRV1 and L. major LRV2. LRV-Lae more closely resembled LRV2. Despite their similar genomic organization, a notable difference was observed in the region where the capsid protein and viral polymerase open reading frames overlap, with a unique -1 situation in LRV-Lae. In vitro infection of murine macrophages showed that LRV-Lae induced a TLR3-dependent inflammatory response as previously observed for LRV1.In this study, we report the presence of an immunogenic dsRNA virus in L. aethiopica human isolates. This is the first observation of LRV in Africa, and together with the unique description of LRV2 in Turkmenistan, it confirmed that LRV was present before the divergence of the L. (Leishmania) and (Viannia) subgenera. The potential implication of LRV-Lae on disease severity due to L. aethiopica infections is discussed

Гуманитарные дисциплины как инструмент формирования учебно-профессиональной самостоятельности студентов негуманитарных вузов

The article deals with the features and capabilities of the humanitarian disciplines as the tool for the formation of students» professional in­dependence. The socio-philosophical and psycho­logical-pedagogical aspects of professional indepen­dence of non-humanitarian universities» students are studied. The problem of the formation of educational and professional independence of students is present­ed at the theoretical and practical level. The research methodology is based on active and interactive ele­ments and forms, as well as the competence-activity approach. The authors refer to the study by S. I. Mo­rozova on the nature, structure, levels of university students» educational and professional independence (EPI) formation. EPI is an integral quality of a stu­dent»s personality, characterized by the ability to or­ganize educational and professional activities aimed at mastering future profession and general cultural competencies on their own. It is considered that qual­itative formation of the general cultural competencies among students of non-humanitarian universities, i.d. the ability to independently acquire knowledge, skills, the need for self-education are possible through hu­manitarian disciplines, organized self-educational activities of the students. Self-educational activities are activities oriented at student focusing on improv­ing their professional competencies and personali­ty as well. The main parameters of self-educational activities include independent search for materials, critical thinking, seeing the problem and solving it, and the self-development ability. Independent stu­dent’s work is considered to be a means of forming student’s interest in continuous self-education. In the article independent work is interpreted as one of the types of the learning process and one of the forms of its organization, aimed at the formation of educa­tional and professional independence of students and contributing to the formation of a creative self-think­ing person, ready for further professional activity and continuous self-education. The authors describe the experience of using interactive and active methods and forms of forming the educational and profession­al independence of students of non-humanitarian uni­versities on the example of historical and philosoph­ical disciplines.В статье анализируются особенности и возможности гуманитарных дисциплин в каче­стве инструмента формирования профессиональ­ной самостоятельности студентов. Исследуются социально-философские и психолого-педагогические аспекты профессиональной самостоятельности студентов негуманитарных вузов. Проблема фор­мирования учебно-профессиональной самостоя­тельности студентов представлена на теорети­ко-практическом уровне. Методология исследования базируется на активных и интерактивных методах и формах и компетентностно-деятельностном под­ходе. Мы ссылаемся на исследование С. И. Морозо­вой о сущности, структуре, уровнях сформированно­сти учебно-профессиональной самостоятельности (УПС) студентов вузов. УПС – это интегральное качество личности студента, характеризующее­ся способностью самостоятельно организовывать свою учебно-профессиональную деятельность, на­правленную на овладение своей будущей профессией и общекультурными компетенциями. Мы считаем, что качественно сформировать у студентов негу­манитарных вузов общекультурные компетенции: способность самостоятельно приобретать знания, умения, навыки, потребность в самовоспитании, самообразовании, способность достигать целей и результатов в учебно-профессиональной деятель­ности, способность к рефлексии, привычку испыты­вать удовольствие от процесса обучения, результа­тов учебно-профессиональной самостоятельности возможно посредством гуманитарных дисциплин, организовав самообразовательную деятельность студентов. Самообразовательную деятельность – это деятельность, в процессе которой студент ориентирован на совершенствование своих профес­сиональных компетенций, а также собственной личности. Основными параметрами самообразова­тельной деятельности являются: самостоятель­ный поиск материалов, их критическое осмысление, выбор между различными точками зрения, или фор­мирование собственного видения и решения про­блемы, способность к саморазвитию. Средством, формирующим интерес студента к постоянному самообразованию, является самостоятельная ра­бота. В нашей статье самостоятельная работа интерпретируется как один из видов процесса об­учения и одна из форм его организации, направлен­ные на формирование учебно-профессиональной самостоятельности студентов и способствующие формированию творческой самостоятельно мысля­щей личности, готовой к последующей профессио­нальной деятельности и непрерывному самообразо­ванию. Изложен опыт применения интерактивных и активных методов и форм формирования учеб­но-профессиональной самостоятельности студен­тов негуманитарных вузов на примере историче­ских и философских дисциплин

Relationship between Helicobacter pylori Virulence Genes and Clinical Outcomes in Saudi Patients

Helicobacter pylori has been strongly associated with gastritis, gastric and duodenal ulcers, and it is a risk factor for gastric cancer. Two major virulence factors of H. pylori have been described: the cytotoxin-associated gene product (cagA) and the vacuolating toxin (vacA). Since considerable geographic diversity in the prevalence of H. pylori virulence factors has been reported, the aim of this work was to determine if there is a significant correlation between different H. pylori virulence genes (cagA and vacA) in 68 patients, from Saudi Arabia, and gastric clinical outcomes. H. pylor was recognized in cultures of gastric biopsies. vacA and cagA genes were detected by polymerase chain reaction (PCR). The cagA gene was obtained with 42 isolates (61.8%). The vacA s- and m- region genotypes were determined in all strains studied. Three genotypes were found: s1/m1 (28%), s1/m2 (40%) and s2/m2 (26%). The s2/m1 genotype was not found in this study. The relation of the presence of cagA and the development of cases to gastritis and ulcer was statistically significant (P < 0.05). The study showed a significant correlation between the vacA s1/m2 genotype and gastritis cases, and a significant correlation between vacA s1/m1 genotype and peptic ulcer cases. The results of this study might be used for the identification of high-risk patients who are infected by vacA s1/m1 genotype of H. pylori strains. In conclusion, H. pylori strains of vacA type s1 and the combination of s1/m1 were associated with peptic ulceration and the presence of cagA gene