[Experimental Evidence of Proliferation and Reproduction of Highly Differentiated Sertoli Cells].

The main results of studies regarding the biology of Sertoli cells under various experimental conditions are considered. Possible potential mechanisms underlying the transition of highly differentiated Sertoli cells to dedifferentiation, limited by proliferation and reproduction and not accompanied by significant phenotypic changes, are discussed.

[Effect of Ultrasmall Gold Nanoparticles on the Murine Native Sperm Chromatin].

The effects of ultrasmall (2-3 nm) gold nanoparticles on native epididymal sperm chromatin of CBAxC57BL/6 hybrid mice and 129/IMG mice with a mutation in the DNA-polymerase iota gene were studied. It is shown that for both mouse strains after sperm incubation in a solution containing Au nanoparticles, at 23, 37 and 60 degrees C for 30 min followed by 1 hour treatment in dithiothreitol solution, a decrease in the number of nuclei with fully decondensed chromatin was observed compared with the control. Though, the manifestation of this effect in the population of 129/IMG mice mature sperm, was weaker. Also we have demonstrated that sperm of both strains that were incubated in a sol of Au nanoparticles at 60 degrees C behave differently under the action of dithiothreitol. A considerable part (-80%) of sperm of CBAxC57BL/6 hybrid mice treated with Au nanoparticles showed high resistance to the action of dithiothreitol, whereas in the case of 129/IMG mice only -30% did, and a partial or complete chromatin decondensation takes place in the remaining sperm. In general, using the method of nuclear chromatin decondensation in vitro for the native sperm, the patterns that we have identified in earlier studies on previously demembranized sperm are confirmed.

Analysis of exhaled breath for diagnosing head and neck squamous cell carcinoma: a feasibility study.

BACKGROUND: Squamous cell carcinoma of the head and neck (HNSCC) are wide-spread cancers that often lead to disfigurement and loss of important functions such as speech and ingestion. To date, HNSCC has no adequate method for early detection and screening. METHODS: Exhaled breath samples were collected from 87 volunteers; 62 well-defined breath samples from 22 HNSCC patients (larynx and pharynx), 21 patients with benign tumours (larynx and pharynx) and 19 healthy controls were analysed in a dual approach: (i) chemical analysis using gas chromatography/mass spectrometry (GC-MS) and (ii) breath-print analysis using an array of nanomaterial-based sensors, combined with a statistical algorithm. RESULTS: Gas chromatography/mass spectrometry identified ethanol, 2-propenenitrile and undecane as potential markers for HNSCC and/or benign tumours of the head and neck. The sensor-array-based breath-prints could clearly distinguish HNSCC both from benign tumours and from healthy states. Within the HNSCC group, patients could be classified according to tumour site and stage. CONCLUSIONS: We have demonstrated the feasibility of a breath test for a specific, clinically interesting application: distinguishing HNSCC from tumour-free or benign tumour states, as well as for staging and locating HNSCC. The sensor array used here could form the basis for the development of an urgently needed non-invasive, cost-effective, fast and reliable point-of-care diagnostic/screening tool for HNSCC.

[Bovine sperm chromatin is not protected from the effects ultrasmall gold nanoparticles].

The response of ejaculated bovine spermatozoa to gold nanoparticles was studied by the standard method of nuclear chromatin decondensation in vitro. After the treatment of semen samples with a hydrosol containing gold nanoparticles with an average diameter of 3.0 nm and a concentration of 1 x 10(15) particles/mL, the ability of sperm nuclei to decondense in the presence of sodium dodecyl sulfate (SDS) and dithiothreitol (DTT) dramatically changed compared to the control. The frequencies of gametes with nondecondensed ("intact"), partially decondensed, and completely decondensed nuclei correlated as 40 : 32 : 28% and 0 : 36 : 64% in the experiment and the control, respectively. Moreover, the appearance of a sufficiently large number of gametes with destructed and almost completely destroyed nuclei was noticed in the spermatozoa treated with gold nanoparticles. This article suggests the putative mechanisms of action of ultrasmall gold nanoparticles on the structural and functional integrity of the deoxyribonucleoprotein (DNP) complex of mature male gametes.

[Effect of gold nanoparticles on mouse spermatogenesis].

The response of the mouse male germ cells exposed to gold nanoparticles (approximately 2.5 nm) was studied. Our investigation demonstrates that treatment with Au nanoparticles for four days does not impair the architecture of the spermatogenic epithelium. Cytogenetic evaluation using micronucleus assay showed that gold nanoparticles can affect the chromosomes of early primary spermatocytes. However, gold nanoparticles did not induce chromosome abnormalities in spermatogonial stem cells. Further, the cauda epididymal sperm was isolated on the 14th day after treatment and was incubated in SDS solution (Na sodium dodecyl) and then in a solution containing DTT (dithiothreitol) to induce nuclear chromatin decondensation. Observations showed that after four days of treatment of spermiogenic (postmeiotic) cells with gold nanoparticles the decondensation process had no differences from the control. On the contrary, in the experiment with the same cells and period of fixation but with a single exposure to gold nanoparticles, the number of mature gametes with totally decondensed nuclei reached 100% as opposed to 44% in the controls.

Direct maxillary irrigation therapy in non-operated chronic sinusitis: a prospective randomised controlled trial.

OBJECTIVE: This study aimed to compare the effectiveness of pharmacological therapy with and without direct maxillary sinus saline irrigation for the management of chronic rhinosinusitis without polyps. METHODS: In this prospective randomised controlled trial, 39 non-operated patients were randomly assigned to be treated with direct maxillary sinus saline irrigation in conjunction with systemic antibiotics and topical sprays (n = 24) or with pharmacological therapy alone (n = 15). Endoscopy, Sino-Nasal Outcome Test and Lund-MacKay computed tomography scores were obtained before, six weeks after and one to two years after treatment. RESULTS: Post-treatment Lund-Mackay computed tomography scores were significantly improved in both cohorts, with no inter-cohort difference identified. Post-treatment nasal endoscopy scores were significantly improved in the study group but were similar to those measured in the control group. The Sino-Nasal Outcome Test-20 results showed improvement in both cohorts, with no difference between treatment arms. CONCLUSION: Maxillary sinus puncture and irrigation with saline, combined with pharmacological treatment improves endoscopic findings in patients with chronic rhinosinusitis without polyps, but has no beneficial effect on symptoms and imaging findings over conservative treatment alone.

Inferior turbinate reduction: comparing post-operative bleeding between different surgical techniques.

OBJECTIVE: Post-operative bleeding is one of the most common and severe complications of turbinate surgery. This study compared post-operative bleeding following partial turbinectomy, submucosal turbinate reduction and endoscopic turbinoplasty. METHODS: Post-operative bleeding was assessed in patients who underwent inferior turbinate intervention by partial turbinectomy, submucosal turbinate reduction or endoscopic turbinoplasty between January 2016 and November 2017 and had completed at least one month of follow up. RESULTS: Of 1035 patients who underwent inferior turbinate surgery during the study period, 751 were included. Of these, 56 (7.5 per cent) presented to the emergency room with post-operative bleeding; 31 (8.4 per cent) had undergone partial turbinectomy, 19 (10.7 per cent) had undergone submucosal turbinate reduction and 6 (3.0 per cent) had undergone endoscopic turbinoplasty. The odds ratio of requiring an intervention to control bleeding was significantly lower in the endoscopic turbinoplasty group than in the submucosal turbinate reduction group (odds ratio = 3.26, 95 per cent confidence interval = 1.02-10.43). CONCLUSION: Endoscopic turbinoplasty had the lowest rate of post-operative bleeding and the lowest rate of patients requiring intervention.

Regeneration potential of transplanted adult mouse sertoli cells.

The regeneration potential of differentiated Sertoli cells subjected to thermal treatment was studied by the method of cell transplantation. Cells from mice with artificial cryptorchism (1.5 months after fixation of the testes in the body) and after culturing (10 days, 37°C) were transplanted. Transplantation of Sertoli cells from 2-3-month-old and 2-day-old mice served as controls. The cells were transplanted into the testes of recipient mice, from which sex cells and Sertoli cells were removed by busulfan and cadmium salt treatment. Adult mouse Sertoli cells exposed to thermal treatment exhibited much higher regeneration potential than intact cells. Two months after transplantation, mature Sertoli cells subjected to thermal treatment populated the recipient testicular tubules, formed new tubules, and in some cases supported the development of sex cells similarly as immature cells from newborn mice.

[Ageing of the spermatogenesis system].

This review summarizes the data characterizing the effect of ageing on the development of male germ cells and their hereditary structures. We have studied causes of spermatogenesis reduction at late stages of ontogenesis. We have focused on age-specific changes of the structural-functional integrity of stem spermatogonial cells and their microenvironment (niche). We also examined several unique and specific features of the spermatogenic system in senescence-accelerated mutant mice (SAM), with accelerated ageing.

[Estimation of the frequencies of induced mutations in spermatogenic cells of senescence-accelerated prone mice of the SAMP1 strain].

The results obtained in this work demonstrate the dynamics of cytogenetic changes of spermatogenic cells in senescence-accelerated prone mice, strain SAMP1, after a single exposure to a chemical mutagen, dipin, at a genetically active dose of 30 mg/kg. In the time interval between days 3 and 28 the frequency of induced spermatogonial micronuclei does not significantly exceed the level of spontaneous mutagenesis. The lack of an experimental effect of micronuclei in this time interval is probably a consequence of mitotic delay and (or) of the death of a considerable part of genetically defective cells in the spermatogonial compartment. Different stages of meiosis exhibit different chemical sensibilities: the yield of round spermatids with micronuclei is maximum after treatment of early primary spermatocytes (preleptotene-leptotene stage) with dipin. The high sensibility of preleptotene and leptotene spermatocytes is confirmed by the sperm head shape abnormality assay. Chromosome damage caused by dipin in spermatogonial stem cells is irreversible, as evidenced by a sharp increase in the frequencies of spermatogonial and meiotic micronuclear aberrations within long periods after treatment. Increased genetic instability in the stem compartment does not lead to irreversible degradation of the system of development of male sex cells in senescence-accelerated SAMP1 mice.

[Response of the spermatogenic system to chemical mutagen dipin in SAMP1 senesce-accelerated mice].

Specific features of spermatogenesis were studied in senesce-accelerated mice of the line SAMP-1 after one-time injection of the chemical mutagen dipin. Quantitative and histomorphological changes in the spermatogenic epithelium proved to develop gradually. Cell loss and disorganization of spermatogenesis reached the peak as late as on days 28 and 35 after the injection. Differentiating spermatogonia manifested increased sensitivity to dipin. In prophase I of meiosis, developing spermatocytes proved to be less sensitive to the cytotoxic action of dipin at the pachytene than at the preleptotene-leptotene stages. Spermatogenesis in most seminiferous tubules was restored by day 56 after dipin treatment. At the end of the experiment (day 100), both quantitative parameters and morphological pattern of spermatogenesis did not differ significantly from those in the control. Thus, the cytotoxic action of dipin does not lead to irreversible structural disorganization of the spermatogenic epithelium in SAMP1 mice. Radioautography revealed a large proportion of highly differentiated Sertoli cells with 3H-thymidine-labeled nuclei in experimental animals. In some cases, structures resembling embryonic seminiferous tubules were revealed in the vicinity of rete testis in testis sections of experimental mice. These structures contained the cells morphologically similar to gonocytes and young Sertoli cells.

[Analysis of spermatogenesis in senescence-accelerated mice].

A comparative analysis of age-related dynamics of spermatogenesis has been performed in mutant mouse lines predisposed or resistant to accelerated senescence (SAMP1 and SAMR1 respectively). The results show that quantitative and morphohistological trends in the development of sperm cells and Sertoli cells in both lines are similar in both lines. Their comparison with data obtained in our previous studies (Zakhidov et al., 2001; Gordeeva et al., 2001) shows that sharp quantitative and qualitative changes in the structure of the spermatogenic system have occurred in senescence-accelerated mice of new generations, which confirms the fact of dynamic instability of the germinal lineage. The role of stem spermatogonial cells in restoration of spermatogenesis in animals reaching the critical age is discussed.

A systematic review of the evidence base for vidian neurectomy in managing rhinitis.

BACKGROUND: Vidian neurectomy has been proposed as a surgical option for rhinitis refractory to medical treatment. However, the evidence base for its benefit remains controversial. METHODS: A systematic review was performed. Studies reporting original data on patients with rhinitis treated by vidian neurectomy were included. Patient-reported outcome measures were the primary outcome investigated; specific peri-operative morbidities were the secondary outcome. RESULTS: A total of 1012 articles fulfilled the search criteria, 32 of which were included in the study. Patient-reported outcome measures were compared before and after surgery in eight studies. There were 529 patients represented in these trials. Significant improvement in rhinorrhoea was reported in all eight studies. Temporary dry eyes was reported in 24.63 per cent of cases (272 out of 1104 cases). There was no report of cranial nerve deficit or eye movement disturbance. CONCLUSION: Endoscopic vidian neurectomy does have a role in the surgical management of refractory rhinitis, particularly in patients with non-allergic rhinitis, but a well-designed cohort trial would be advantageous to clarify long-term outcomes.

An attempt to influence DNA content in postmitotic Purkinje cells of the cerebellum.

Feulgen-DNA content has been measured cytophotometrically in granule and Purkinje cells of the rat and mouse cerebellum. The study has confirmed that under normal conditions a very small part of the P. cell population in rats (less than 3%) possesses a Feulgen-DNA surplus ranging from 2C to 4C. In mice, the hyperdiploid (H2C) P. cells are even more rare. The occurrence of H2C-P. cells in rats and probably also in mice has not been substantially changed in animals exposed to factors interfering with chromatin structure and/or its template activity; the H2C-P. cells seem to be slightly more frequent after injecting mice with corticoids (Urbason) or in animals suffering from ectromelia or hereditary Purkinje cell degeneration. The incidence of H2C-P. cells has neither been substantially affected by experimental conditions which are known to lead to functional and/or metabolic stimulation of the cerebellum. Functional changes in the number of H2C-P. cell nuclei may, however, be short-term or transient in character and therefore might have excaped detection in our models. The findings rule out an impact of some reasons suspected for the artefactual origin of Feulgen H2C DNA values as e.g. compactness of the chromatin.

[Ultrastructure and the number of nucleoli in the neurons of the rat cerebellar cortex]. / Ul'trastruktura i chislo iadryshek v neironakh kory mozzhechka krysy.

The ultrastructure and number of nucleoli in rat cerebellar nervous cells (the Purkinje and granule cells) were studied using the silver staining procedure and serial ultrathin sections. In the Purkinje cells one large nucleonemal nucleolus about 4 mcm in diameter was usually observed; fibrillar centers are weakly expressed. About 6% of the Purkinje cells had two nucleoli different in size. In granule cells 2-3 nucleoli about 1 mcm in diameter were detected. They contained a distinct fibrillar center and a dense fibrillar RNP-component, RNP-granules were barely visible. These data show that the Purkinje cells are more active transcriptionally than granule cells. The distinctions in the ultrastructure and number of nucleoli in the Purkinje and granule cells correspond to an earlier reported hypothesis about the intra- and interpopulation heterogeneity of cerebellar nervous cells for topography of ribosomal genes.

[Number of Purkinje cells with an increased DNA content in rat cerebellum]. / Kolichestvo kletok Purkin'e s uvelichennym soderzhaniem DNK v mozzhechke krysy.

Using cytospectrophotometry, DNA quantities were measured in the Purkinje cells from a large number of animals. The Purkinje cells appeared to be either diploid or having hypercontent of DNA (HC). HC-nuclei were detected in the cerebellum of rats of different lines; the number of nuclei not depending of the animal's sex or age. No difference in the number of HC-nuclei was recorded in the animals of different litters. Out of 45 Wistar and non-inbred rats, HC-nuclei was found in 19 animals (0.5 to 7.0 per animal).

[The possible cytophotometry of nucleolar proteins. The prospects for research on the status of the nucleolus apparatus]. / Vozmozhnost' tsitofotometrii iadryshkovykh belkov. Perspektivy issledovanii sostoianiia iadryshkovogo apparata.

Possibility of cytophotometry application was established for the acid nucleolar proteins reacting with silver nitrate. The increase in transcription in the Purkinje neurons results in the expansion of the Ag-proteins areas, whereas their amount may be not increased. The amount and area of the Ag-proteins double in hepatocytes through polyploidization, but the number of nucleoli does not correspond to the gene dosage. This lack of correspondence was also revealed in micronucleoli. The number of nucleoli is not similar in the nuclei of some binuclear hepatocytes, and this disproportionality increases through polyploidization.

[Intra- and interpopulation heterogeneity of the cerebellar nerve cells in topography and ribosomal gene content]. / Vnutri- i mezhpopuliatsionnaia geterogennost' nervnykh kletok mozzhechka po topografii i soderzhaniiu ribosomnykh genov.

Using the method of in situ hybridization of ribosomal DNA with DNA of isolated nuclei, a study was made of localization and relative content of ribosomal genes in the Purkinje cells and other cells of the rat cerebellum. It has been shown that various cells of cerebellum have, on the average, the same number of ribosomal genes. A subpopulation with amplified ribosomal genes have been found among the nuclei of Purkinje's cells.

[Ribosomal RNA synthesis and the count and type of nucleoli in rat hepatocytes]. / Sintez ribosomnoi RNK, chislo i tip iadryshek v gepatotsitakh krysy.

Changes in the activity and number of nucleoli has been studied in hepatocytes of the same rat before and after partial hepatectomy. In the latter case, the rate of ribosomal RNA (rRNA) synthesis was increased by a factor of 1.7, on the average. The synthesis was evaluated by incorporation of adenosine triphosphate in situ after alpha-amanitine treatment of squashed cells. In 2c, 4c and 8c nuclei, the number of labeled nucleoli was less than in nucleolonemic nucleoli stained with silver nitrate. These values were drawn together during stimulation of rRNA synthesis but the difference was often preserved especially for highly ploid nuclei. The lack of activity in part of nucleolonemic nucleoli was assumed. After stimulation of RNA synthesis, the number of nucleolonemic and satellite nucleoli decreased, while the size of preserved nucleoli was enlarged. The number of nucleoli did not correspond to the nuclear ploidy; their total size was proportional to the gene dosage. The mean level of rRNA synthesis was similar in nuclei of the same ploidy with two, three and four nucleoli. A great variability was detected in the rate of rRNA synthesis as well as in the number of nucleoloneme and satellite nucleoli in hepatocytes of the control and experimental rats. Some difference was observed in the label degree for nucleoli of the same nucleus.

[Oogenesis in hydra. II. Cytophotometric determination of DNA concentration in the nuclei of somatic and sex cells]. / Issledovanie oogeneza u gidry. Soobshchenie II. Tsitofotometricheskoe opredelenie soderzhaniia DNK v iadrakh polovykh i somaticheskikh kletok

During sexual reproduction in Hydra, interstitial cells in the female sex zone of the body (i-cells) undergo mitotic division and form a thickening in the epiderm. The proliferation of i-cells is accompanied by the increase of cytoplasm volume and by the appearance in the cytoplasm of a great number of membranous structures (rough endoplasmic reticulum, Golgi apparatus and mitochondria), enzymatic granules, lipid inclusions and glycogen. All cells of the epidermal thickening soon (in approximately twenty four hours) acquire the characteristics of typical phagocytes. However it is the cell situated inside the group of syncytially connected ones and adjacent to mesogloea that begins to grow rapidly and phagocytize surrounding cells. The cells of the epidermal thickening, though they are often given the name of oogonia, were found to have a tetraploid DNA content in their nuclei. The presence of four unseparated centrioles of equal size suggests that all preparatory processes for division were completed. A conclusion was drawn that cells of the epidermal thickening undergo premeiotic DNA synthesis prior to their phagocytizing by the growing oocyte and, thus, are oocytes themselves. The oogonial stage in Hydra coincides with the early period of mitotic reproduction of i-cells. The data obtained are discussed from the viewpoint of the formation of the accessory gonad apparatus.