HPV genotypes in the oral cavity/oropharynx of children and adolescents: cross-sectional survey in Poland

Human papillomaviruses (HPVs) are a very complex group of pathogenic viruses, with more than 80 types, causing human infection. Given the prevalence of HPV infection and its relationship with the development of cervical and many other cancers, HPV vaccine development has been a major public health initiative worldwide in the last decade. The aim of the presented study was to identify HPV DNA by MY-PCR in 4,150 school children and adolescents, aged 10–18 years in the Wielkopolska region, Poland. All individuals were asked to fill in extensive questionnaires; further normal, oral squamous cells were collected from each pupil. Cellular DNA was isolated and used as a MY-PCR template to estimate the incidence of HPV-active infection. Forty five subjects (1.08% of the sample) were carriers of oropharyngeal HPVs. HPV status and variables of interest, such as age, gender, socioeconomical status, and risk factors (smoking and sexual intercourse history, alcohol consumption) were not correlated. The presence of HPVs in the oral cavity was cumulated in several schools of the region. DNA sequencing of MY-PCR products revealed only four HPV genotypes. The most frequent genotype was HPV11 (38/45 HPV-positive cases), while other more rare genotypes were HPV6 (3/45), HPV12 (3/45), and HPV57 (1/45). Conclusion: Our findings presented herein, reveal a relatively low prevalance of oropharyngeal HPVs in Polish adolescents and fill an important gap in the knowledge of oral HPV infections of children above 10 years and adolescents

Cholinergic Agonists and Antagonists Have an Effect on the Metabolism of the Beetle <i>Tenebrio Molitor</i>

Synthetic insecticides are still widely used in plant protection. The main target for their action is the nervous system, in which the cholinergic system plays a vital role. Currently available insecticides have low selectivity and act on the cholinergic systems of invertebrates and vertebrates. Acetylcholine, a cholinergic system neurotransmitter, acts on cells by two types of receptors: nicotinic and muscarinic. In mammals, the role of muscarinic acetylcholine receptors (mAChRs) is quite well-known but in insects, is still not enough. Based on data indicating that the muscarinic cholinergic system strongly affects mammalian metabolism, we investigated if it similarly occurs in insects. We investigated the influence of agonists (acetylcholine, carbachol, and pilocarpine) and antagonists (tropane alkaloids: atropine and scopolamine) of mAChRs on the level of selected metabolites in Tenebrio molitor beetle trophic tissues. We analyzed the glycogen content in the fat body and midgut, the total free sugar concentration in the hemolymph and the lipid amount in the fat body. Moreover, we analyzed the levels of insulin-like peptides in the hemolymph. The tested compounds significantly influenced the mentioned parameters. They increased the glycogen content in the fat body and midgut but decreased the concentration of free sugars in the hemolymph. The observed effects were tissue-specific, and were also time- and dose-dependent. We used nonligated and neck-ligated larvae (to eliminate the influence of head factors on tissue metabolism) to determine whether the observed changes are the result of direct or indirect impacts on tissues. The obtained data suggest that the cholinergic system affects the fat body and midgut indirectly and directly and a pleiotropic role for mAChRs exists in the regulation of energy metabolism in insects. Moreover, tested compounds significantly affected the level of insulin-like peptides in hemolymph. Our studies for the first time showed that mAChRs are involved in regulation of insect metabolism of trophic tissues, and act on them directly and indirectly. Improved knowledge about insect cholinergic system may help in searching more selective and environment-friendly solutions in pest management

Neuropeptidomes of Tenebrio molitor L. and Zophobas atratus Fab. (Coleoptera, Polyphaga: Tenebrionidae)

Neuropeptides are signaling molecules that regulate almost all physiological processes in animals. Around 50 different genes for neuropeptides have been described in insects. In Coleoptera, which is the largest insect order based on numbers of described species, knowledge about neuropeptides and protein hormones is still limited to a few species. Here, we analyze the neuropeptidomes of two closely related tenebrionid beetles: Tenebrio molitor and Zophobas atratus-both of which are model species in physiological and pharmacological research. We combined transcriptomic and mass spectrometry analyses of the central nervous system to identify neuropeptides and neuropeptide-like and protein hormones. Several precursors were identified in T. molitor and Z. atratus, of which 50 and 40, respectively, were confirmed by mass spectrometry. This study provides the basis for further functional studies of neuropeptides as well as for the design of environmentally friendly and species specific peptidomimetics to be used as biopesticides. Furthermore, since T. molitor has become accepted by the European Food Safety Authority as a novel food, a deeper knowledge of the neuropeptidome of this species will prove useful for optimizing production programs at an industrial scale

Owady - alternatywne organizmy modelowe do badań chorób człowieka

W ostatniej dekadzie nastąpił gwałtowny wzrost zainteresowania wykorzystaniem bezkręgowców, w tym owadów, jako organizmów modelowych w badaniach chorób człowieka. Opublikowano liczne prace, w których wykazano możliwość wykorzystania owadzich modeli w badaniach chorób neurodegeneracyjnych, cukrzycy, otyłości, czy chorób serca. Szybki rozwój technik biologii molekularnej, biotestów fizjologicznych i farmakologicznych, a także poznanie genomów muszki Drosophila melanogaster, chrząszcza Tribolium castaneum, jedwabnika Bombyx mori i pszczoły Apis mellifera dodatkowo stanowią silne wsparcie metodologiczne tego kierunku badań. Poznano szereg genów owadów, będących ortologami genów ludzkich, odpowiedzialnych za rozwój różnych chorób, zidentyfikowano wiele białek kodowanych przez te geny, scharakteryzowano ich fenotypy morfologiczne i fizjologiczne, a także opisano działanie u owadów niektórych leków stosowanych w chorobach neurodegeneracyjnych i kardiologicznych człowieka.Over the last decade the interest of using invertebrates, including insects, as model organisms in studies of human diseases has rapidly increased. Until now, hundreds of papers which demonstrate the possibility of using insect models in studies on human neurodegenerative diseases, diabetes, obesity or heart diseases were published. The rapid development of molecular biology techniques, physiological and pharmacological bioassays, as well as genome sequencing of fruit fly Drosophila melanogaster, red flour beetle Tribolium castaneum, silkworm Bombyx mori and honey bee Apis mellifera also provides a strong methodological support for this research approach. As a result of these studies a number of insect genes, orthologs of human genes known to be responsible for the development of various diseases, and many proteins encoded by these genes were identified. Also morphological and physiological phenotypes of different genes and the action of some drugs applied in neurodegenerative and cardiac diseases in humans have been characterized in insects

The expression of RARs and RXRs in cervical cancer tissues HPV(+)

Introduction: Retinoids are essential for the maintenance of epithelial differentiation and play a fundamental role in chemoprevention of epithelial carcinogenesis. Retinoids exert their biological functions through nuclear receptors, retinoic acid receptor (RAR) and retinoid X receptor (RXR). The present study made an effort to analyze serum blood concentration of retinoids in women with cervical cancer as well as to assess an expression of RARs and RXRs in postoperative cervical cancer tissues HPV 16/18 positive. Material and methods: The study material included tissue samples of 42 squamous cell carcinoma, 7 samples of adenocarcinoma cervix uteri and 26 samples carcinoma in situ. The assessment of serum level of retinol, expression of retinoid receptors and presence of HPVs genome was performed. Results and conclusions: The mean retinol content in blood serum of patients with cervical cancer associated with HPV infection type 16 and/or 18 was lower than in controls. The level of RARα, RARβ and RXRβ mRNA expression was significantly decreased in the study group of women with CIS, squamous cell carcinoma and adenocarcinoma cervix to compare morphologically to the control women. In squamous cell carcinoma and carcinoma in situ were found to exhibit a decreased expression of RARα by about 75%, RARβ by 90%, RXRβ by 70% and 83% respectively compared to the control tissues. Among adenocarcinomas RARα, RARβ, RXRβ were expressed in 10%. In the study cancer tissues RARγ, RXRα and RXRγ were expressed on the same level as in the control tissues.Wprowadzenie: Retinoidy są ważnymi czynnikami biorącymi udział w regulacji różnicowania i proliferacji komórek nabłonkowych, a także odgrywają ważną rolę w ich chemioprofilaktyce i nowotworzeniu. Retinoidy wywierają swój efekt biologiczny poprzez receptory jądrowe RAR i RXR. Cel pracy: Celem przeprowadzonych badań była analiza stężenia retinolu w surowicy krwi kobiet z rakiem szyjki macicy oraz ocena ekspresji mRNA RAR i RXR w komórkach raka szyjki macicy zakażonych wirusem brodawczaka ludzkiego (HPV) typu 16. i 18. Materiał i metoda: Materiał badany obejmował 42 skrawki pooperacyjne tkanek raka płaskonabłonkowego, 7 skrawków gruczolakoraka szyjki macicy, a także 26 skrawków raka przedinwazyjnego. U osób, od których pochodziły skrawki tkanek, oceniono surowiczy poziom retinolu, a w tkankach ekspresję receptorów retinoidowych na poziomie mRNA i obecność sekwencji genomu HPV z zastosowaniem technik PCR. Wyniki i wnioski: Średnia zawartość retinolu w surowicy krwi pacjentek z rakiem szyjki macicy z towarzyszącym zakażeniem wirusem HPV typu 16. i 18. była niższa niż w grupie kontrolnej, ale nie znamienna statystycznie. Poziom RARα, RARβ oraz ekspresja mRNA dla RXRβ były znamiennie niższe w grupie badanej – u pacjentek z rakiem przedinwazyjnym (CIS), z rakiem płaskonabłonkowym i z gruczolakorakiem szyjki macicy, w porównaniu z kobietami z grupy kontrolnej. W przypadku raka płaskonabłonkowego i raka przedinwazyjnego stwierdzono zmniejszenie ekspresji RARα mniej więcej o 75%, RARβ o 90%, a RXRβ odpowiednio o 70% i 83% w porównaniu z komórkami tkanek kontrolnych. W przypadku gruczolakoraków ekspresja RARγ, RXRα and RXRγ była na poziomie takim jak w tkankach grupy kontrolnej

Beetles as Model Organisms in Physiological, Biomedical and Environmental Studies – A Review

Model organisms are often used in biological, medical and environmental research. Among insects, Drosophila melanogaster, Galleria mellonella, Apis mellifera, Bombyx mori, Periplaneta americana, and Locusta migratoria are often used. However, new model organisms still appear. In recent years, an increasing number of insect species has been suggested as model organisms in life sciences research due to their worldwide distribution and environmental significance, the possibility of extrapolating research studies to vertebrates and the relatively low cost of rearing. Beetles are the largest insect order, with their representative – Tribolium castaneum – being the first species with a completely sequenced genome, and seem to be emerging as new potential candidates for model organisms in various studies. Apart from T. castaneum, additional species representing various Coleoptera families, such as Nicrophorus vespilloides, Leptinotarsa decemlineata, Coccinella septempunctata, Poecilus cupreus, Tenebrio molitor and many others, have been used. They are increasingly often included in two major research aspects: biomedical and environmental studies. Biomedical studies focus mainly on unraveling mechanisms of basic life processes, such as feeding, neurotransmission or activity of the immune system, as well as on elucidating the mechanism of different diseases (neurodegenerative, cardiovascular, metabolic, or immunological) using beetles as models. Furthermore, pharmacological bioassays for testing novel biologically active substances in beetles have also been developed. It should be emphasized that beetles are a source of compounds with potential antimicrobial and anticancer activity. Environmental-based studies focus mainly on the development and testing of new potential pesticides of both chemical and natural origin. Additionally, beetles are used as food or for their valuable supplements. Different beetle families are also used as bioindicators. Another important research area using beetles as models is behavioral ecology studies, for instance, parental care. In this paper, we review the current knowledge regarding beetles as model organisms and their practical application in various fields of life science