The impact of environmental and behavioural cofactors on the development of cervical disorders in HR-HPV-infected women in Serbia.

Persistent infection with one or more highly oncogenic human papillomaviruses (HPVs) or high-risk-HPV (HR-HPV) is necessary but not a sufficient aetiological agent for the development of cervical neoplasia. A number of viral, host, environmental and behavioural factors are suggested to be associated with the progression of cervical disorder. This study aimed to evaluate the impact of environmental and behavioural cofactors on the development of cervical disorders in HR-HPV-infected women in Serbia. A total of 541 women have been tested by PCR for the presence of HPV on the cervix. HPV genotypes were determined by direct DNA sequencing. Women identified as HR-HPV-positive were further classified into four subgroups according to their cytological status. All relevant information about demographical and behavioural factors was obtained by interviewer-based questionnaire. A number of analytical and descriptive statistical methods were used for processing the data. The cofactors found to be of significance for the progression of cervical disease were older age, body mass index >25, lower educational level, long-term smoking, previous genital infections and cervical interventions. On the other hand, condom use was found to have a protective role. Information about these cofactors might be very important for the development of more efficient cancer prevention programmes and promotion of anti-HPV vaccination.

Endovascular Management of Acute Stroke in the Elderly: A Systematic Review and Meta-Analysis.

BACKGROUND: Acute ischemic stroke occurs more frequently, presents with more severe symptoms, and has worse outcomes in elderly patients. The safety and efficacy of endovascular therapy for acute stroke in this age group has not been fully established. PURPOSE: We present the results of a systematic review and meta-analysis examining clinical, procedural, and radiologic outcomes of endovascular therapy for acute stroke in patients older than 80 years of age. DATA SOURCES: We searched PubMed, MEDLINE, and EMBASE from 1992 to week 35 of 2017 for studies evaluating endovascular therapy for acute stroke in the elderly. STUDY SELECTION: Two independent reviewers selected studies and abstracted data. The primary end point was good functional outcome at 3 months defined as modified Rankin Scale score of ≤2. DATA ANALYSIS: Data were analyzed using random-effects meta-analysis. DATA SYNTHESIS: Seventeen studies reporting on 860 patients were included. The rate of good functional outcome at 3 months was 27% (95% CI, 21%-32%). Mortality at 3 months was 34% (95% CI, 23%-44%). Successful recanalization was achieved in 78% of patients (95% CI, 72%-85%). Procedure-related complications occurred in 11% (95% CI, 4%-17%). The incidence of intracranial hemorrhage was 24% (95% CI, 15%-32%), and for symptomatic intracranial hemorrhage, it was 8% (95% CI, 5%-10%). The mean time to groin was 251 minutes (95% CI, 224-278 minutes). Procedure time was 99 minutes (95% CI, 67-131 minutes). LIMITATIONS: I2 values were above 50% for all outcomes, indicating substantial heterogeneity. CONCLUSIONS: Good functional recovery in octogenarians treated with endovascular therapy for acute stroke can be achieved in a high proportion of patients despite the higher incidence of comorbidity in this cohort. Outcomes are inferior to those reported for younger patients; however, endovascular therapy can allow at least 1 in 4 patients older than 80 years of age to regain independent function at 3 months. More research is required to improve patient selection in the elderly, but age should not be a discriminator when deciding to offer endovascular therapy for patients with acute stroke.

Specific detection of GII-1 lineage of infectious bronchitis virus.

Infectious bronchitis virus (IBV) is a worldwide prevalent RNA virus that causes highly contagious and economically devastating disease in chicken. The virus exists in many different genetic forms which made the disease control very difficult. The present study describes the development and validation of TaqMan probe-based real-time reverse transcription-polymerase chain reaction (real-time RT-PCR) targeting the S1 coding region of S gene characteristic for the GII-1 lineage (formerly the D1466-like variant) of IBV. These strains are quite different from other European IBV belonging to different lineages of the GI genotype. The developed method was 30-fold more sensitive than used so far for standard nested RT-PCR with detection limit of 56 RNA copies per reaction. The specificity of the assay was also evaluated with a panel of different poultry pathogens. Repeatability and reproducibility of the method was very high with coefficients of variation lower than 4%. One hundred and twenty-seven IBV-positive samples were tested by this method and GII-1 strains were detected in four of them (3·15%) which indicate a decrease in the GII-1 IBV prevalence in Poland. The assay was proven to be a valuable tool for rapid diagnosis of GII-1 lineage of IBV strains and moreover it enabled the monitoring of viral loads which can be used to assess disease progression. SIGNIFICANCE AND IMPACT OF THE STUDY: This study reports a TaqMan probe-based real-time reverse transcription-polymerase chain reaction (real-time RT-PCR) for rapid and accurate identification of GII-1 lineage (formerly D1466-like variant) of infectious bronchitis virus (IBV). The assay revealed to be more sensitive than standard nested RT-PCR assay, previously used for this purpose. The developed assay has been tested on numerous field samples and revealed 3·15% prevalence of this lineage of IBV in Polish chicken population. Moreover, this new assay enables the assessment of viral load measurement which might be useful for epidemiology and pathogenesis studies.

Cross-sectional survey of selected enteric viruses in Polish turkey flocks between 2008 and 2011.

BACKGROUND: Enteric diseases are an important health problem for the intensive poultry industry, resulting in considerable economic losses. Apart from such microbiological agents associated with enteritis as bacteria and parasites, a lot of research has been recently conducted on viral origin of enteric diseases. However, enteric viruses have been identified in intestinal tract of not only diseased but also healthy poultry, so their role in enteritis is still unclear. The present study aimed at determination of the prevalence of four enteric viruses, namely astrovirus, coronavirus, parvovirus and rotavirus in meat-type turkey flocks in Poland as well as at statistical evaluation of the occurrence of the studied viruses and their relationships with the health status and the age of birds. Two hundred and seven flocks of birds aged 1-20 weeks originating from different regions of the country were investigated between 2008 and 2011. Clinical samples (10 individual faecal swabs/flock) were duly processed and examined using molecular methods targeting the conservative regions of viral genomes: RNA-dependent RNA polymerase gene of astrovirus, non-structural 1 gene of parvovirus, non-structural protein 4 gene of rotavirus, and 5' untranslated region fragment of turkey coronavirus. Different statistical methods (i.e. the independence chi-square test, the correspondence analysis and the logistic regression model) were used to establish any relationships between the analyzed data. RESULTS: Overall, 137 (66.2%, 95% CI: 59.3-72.6) of the 207 turkey flocks sampled were infected with one or more enteric viruses. Among the 137 flocks, 74 (54%, 95% CI: 45.3-62.6) were positive for one virus, whereas 54 (39.4%, 9 5% CI: 31.2-48.1) and 9 (6.6%, 95% CI: 3.1-12.1) were co-infected with two or three different enteric viruses, respectively. No flock was simultaneously infected with all four viruses studied. The prevalence of astrovirus infection was 44.9% (95% CI: 38.0-52.0), parvovirus 27.5% (95% CI: 21.6-34.2), rotavirus 18.8% (95% CI: 13.8-24.8), and coronavirus 9.7% (95% CI: 6.0-14.5). Young turkeys aged 1-4 weeks old had the highest (82.1%, 95% CI:71.7-89.8) prevalence of viral infection. Applied statistical methods have indicated the dependence of rotavirus infection as well as the co-infection with multiple viruses and the health status of turkeys. Furthermore, our results statistically confirm that especially young birds are susceptible to infection with rotavirus and astrovirus. CONCLUSIONS: The study demonstrated the presence of astrovirus, coronavirus, parvovirus and rotavirus infections in Polish turkey farms. These viruses were detected in both healthy and diseased birds. However, the presented results provide valuable feedback which could help to evaluate the role of some enteric viruses in the etiology of enteritis in turkey.

Red blood cell ω-3 fatty acid levels and markers of accelerated brain aging.

OBJECTIVE: Higher dietary intake and circulating levels of docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) have been related to a reduced risk for dementia, but the pathways underlying this association remain unclear. We examined the cross-sectional relation of red blood cell (RBC) fatty acid levels to subclinical imaging and cognitive markers of dementia risk in a middle-aged to elderly community-based cohort. METHODS: We related RBC DHA and EPA levels in dementia-free Framingham Study participants (n = 1575; 854 women, age 67 ± 9 years) to performance on cognitive tests and to volumetric brain MRI, with serial adjustments for age, sex, and education (model A, primary model), additionally for APOE ε4 and plasma homocysteine (model B), and also for physical activity and body mass index (model C), or for traditional vascular risk factors (model D). RESULTS: Participants with RBC DHA levels in the lowest quartile (Q1) when compared to others (Q2-4) had lower total brain and greater white matter hyperintensity volumes (for model A: ß ± SE = -0.49 ± 0.19; p = 0.009, and 0.12 ± 0.06; p = 0.049, respectively) with persistence of the association with total brain volume in multivariable analyses. Participants with lower DHA and ω-3 index (RBC DHA+EPA) levels (Q1 vs. Q2-4) also had lower scores on tests of visual memory (ß ± SE = -0.47 ± 0.18; p = 0.008), executive function (ß ± SE = -0.07 ± 0.03; p = 0.004), and abstract thinking (ß ± SE = -0.52 ± 0.18; p = 0.004) in model A, the results remaining significant in all models. CONCLUSION: Lower RBC DHA levels are associated with smaller brain volumes and a "vascular" pattern of cognitive impairment even in persons free of clinical dementia.

Evidence of persistence and multiple genetic modifications of H7N7 low-pathogenic avian influenza virus in wild mallards in Poland provided by phylogenetic studies.

Genetic characterization of the whole genome of four avian influenza H7N7 viruses isolated in three successive winter seasons (2007 to 2009) from wild mallards in three cities in Poland was performed. All of the tested strains were of low pathogenicity and no molecular marker associated with an increased adaptation to poultry, mammals or resistance to antiviral drugs was found. The major outcome of the phylogenetic studies was that the isolate A/mallard/Poland/446/09 (detected in December 2009) shared a recent common ancestor with A/mallard/Poland/41/09 (isolated in February 2009) in relation to HA and PB1 genes, with A/mallard/Poland/16/09 (found in January 2009) regarding NA and NS genes, and with A/mallard/Poland/01/08 (recovered in December 2007) as regards the NS gene. Interestingly, A/mallard/Poland/16/09 and A/mallard/Poland/446/09 were isolated at the same sampling site almost exactly 1 year apart, which points to resident population of mallards (and other resident waterfowl) as responsible for the perpetuation of avian influenza virus (AIV) in the given area between successive winters. On the other hand, the ornithological data discussed in detail in the paper strongly suggest that the virus transmission between close sites but located in different urban areas is most probably achieved by migratory birds, a fact additionally supported by a close relatedness between different gene segments of Polish H7N7 and AIV detected in wild birds in Europe. A high heterogeneity of the gene pool found in the study is indicative of frequent reassortment events. Additionally, two H7N7 isolates were shown to possess selected genes closely related to AIV detected in domestic poultry in Italy and the Czech Republic. The present study corroborates the importance of active surveillance in wild birds as a valuable tool for early warning of avian influenza in poultry.

Association of parental dementia with cognitive and brain MRI measures in middle-aged adults.

OBJECTIVES: Studies of autosomal dominant Alzheimer disease (AD) have shown structural and cognitive changes in mutation carriers decades prior to clinical disease. Whether such changes are detectable in offspring of persons with sporadic dementia remains unknown. We related prospectively verified parental dementia to brain MRI and cognitive testing in the offspring, within a 2-generational community-based cohort. METHODS: A total of 717 Framingham offspring (mean age: 59 +/- 8 years) were studied. In multivariate analyses, we compared offspring with and without verified parental dementia (and AD) for 1) performance on tests of memory, abstract reasoning, and cognitive flexibility, and 2) volumetric brain MRI measures of total cerebral brain volume (TCBV), hippocampal volume (HV), and white matter hyperintensity volume (WMHV), assessed cross-sectionally and longitudinally. RESULTS: When testing the association of parental dementia and AD with baseline cognitive performance, we observed an interaction of parental dementia and AD with APOE epsilon4 status (p < 0.002). In APOE epsilon4 carriers only (n = 165), parental dementia was associated with poorer scores on tests of verbal memory (beta = -1.81 +/- 0.53, p < 0.001) and visuospatial memory (beta = -1.73 +/- 0.47, p < 0.001). These associations were stronger for parental AD (beta = -1.97 +/- 0.52, p < 0.001, beta = -1.95 +/- 0.48, p < 0.001), equivalent to 14-16 years of brain aging. Among APOE epsilon4 carriers, offspring of participants with dementia were also more likely to show an annual decline in TCBV in the top quartile (odds ratio = 4.67 [1.26-17.30], p = 0.02). Regardless of APOE epsilon4 status, participants with parental dementia were more likely to be in the highest quartile of decline in executive function test scores (odds ratio = 1.61 [1.02-2.53], p = 0.04). CONCLUSIONS: Among middle-aged carriers of the APOE epsilon4 allele, parental dementia and Alzheimer disease were associated with poorer verbal and visuospatial memory and a higher rate of global brain atrophy.

Identification of senescence-associated genes from daylily petals.

The petals of daylily (Hemerocallis hybrid) have a genetically based program that leads to senescence and cell death ca. 24 h after the flower opens. In order to determine the components of this program, six cDNAs, whose levels increase during petal senescence, were isolated and sequenced and designated DSA3, 4, 5, 6, 12 and 15. All six DSAs are members of gene families and all but DSA5 and DSA6 have one to three other very similar genes. GenBank database homology searches indicate that DSA3 is most similar at the amino acid level to an in-chain fatty acid hydroxylase which is bound to cytochrome P450, DSA4 may be an aspartic proteinase, DSA5 is as yet unidentified, DSA6 is a putative S1-type nuclease, DSA12 is very similar to a cytochrome P450-containing allene oxide synthase, and DSA15 may be a fatty acid elongase. Except for DSA12, the genes are expressed at low levels in daylily roots. Levels of the DSA mRNAs in leaves are less than 4% of the maximum detected in petals, and there are no clear differences between younger and older leaves. With the exception of DSA4, accumulation of the DSA mRNAs is increased 3.2 to 43 times by a concentration of abscisic acid that causes premature senescence of the petals. The relationship of the putative DSA gene products to senescence and cell death of daylily petals is discussed.

Distribution and reciprocal interactions of 3H-melatonin and 125I-thyroxine in peripheral, neural, and endocrine tissues of bullfrog tadpoles.

Tissue distribution of 125I-thyroxine (T4) and 3H-melatonin and the effect of each hormone on the tissue content of the other were studied because previous work indicated that melatonin antagonized metamorphosis through peripheral, as well as thyroidal effects. Late pre- to prometamorphic Rana catesbeiana tadpoles on an 18 light:6 dark cycle were used for injection of hormones in vivo or to supply tissues for in vitro hormone administration. Labeled melatonin uptake was highest in intestine, ventral skin and pituitary; lowest in thyroid and brain and intermediate in hindlimb, tail and gills. The tissue content of labeled T4 was distributed in nearly the same way, except that the thyroid level was relatively higher, and pituitary lower, than that of labeled melatonin. The pineal, studied only in the tracer T4 experiments, had the highest content of labeled T4 of all tissues. Simultaneous injection of either 0.007 or 0.2 microgram T4 increased 3H-melatonin uptake into peripheral tissues that undergo major metamorphic changes but not into neural or endocrine organs. In contrast, 0.033, 3.75 or 15 micrograms melatonin had no significant influence on the content of 125I-T4 in any tissue studied in vivo. Results of in vitro labeling of selected tissues were generally in agreement with the in vivo work except that the 125I-T4 content of intestinal segments from late prometamorphic larvae was lower in melatonin-treated than in control groups. The results suggest that peripheral tissues are a major site for T4-melatonin interactions and that T4 may modulate its own action through influencing melatonin levels in target tissues and perhaps in the thyroid. Because melatonin had no effect on tissue T4 content in young tadpoles, retardation of metamorphic events by melatonin does not seem to involve modulation of T4 availability to the tissues.

Effect of melatonin on the response of the thyroid to thyrotropin stimulation in vitro.

Thyroidal-melatonin interactions are of particular importance to amphibian development since the thyroid controls the progress of metamorphosis while melatonin may coordinate its rate with prevailing environmental conditions. Melatonin antagonized thyroxine (T4) action at the tissue level and directly inhibited baseline T4 secretion in culture, so the present work sought to determine if it antagonized the response of the thyroid to thyroid stimulating hormone (TSH) as well. A preliminary experiment showed that, in Rana pipiens, the concentration of TSH (0.2 microg/ml) used in the culture of tadpole thyroids stimulated T4 secretion as much as frog pituitaries, but more than late premetamorphic tadpole pituitaries. There was no significant effect of 1 to 15 microg/ml melatonin in TSH-containing thyroid cultures of various Rana species of tadpoles and frogs in experiments with media collected once every 24 or 48 hr, although 15 microg/ml melatonin tended to depress T4 secretion. In a final experiment, a higher melatonin concentration was used as well as more frequent media collections. Thyroids from prometamorphic Rana catesbeiana tadpoles were cultured in L-15 media with periodic stimulation by 0.2 microg/ml TSH, or TSH and 10 or 100 microg/ml melatonin. Media were collected at the end of two 3-hr TSH pulses, and every 8 hr thereafter for the next 3 days. Melatonin was administered until the end of Day 2 while TSH was given only on Day 2 in addition to the original 3-hr pulses. The secretion of T4 was inhibited significantly by 10 microg/ml melatonin at only two of the early media collections. In contrast, 100 micro;g/ml melatonin significantly depressed T4 secretion in response to TSH at all but one interval and completely inhibited the thyroidal response to TSH reintroduced into the media on Day 2. The findings suggest that a high concentration of melatonin is inhibitory to the thyroidal response to TSH, but that lower concentrations do not significantly overcome the TSH stimulus.

Effect of melatonin on the anuran thyroid gland: follicle cell proliferation, morphometry, and subsequent thyroid hormone secretion in vitro after melatonin treatment in vivo.

The thyroid gland controls the progress of metamorphosis, although other hormones influence metamorphic rate, including melatonin, which may coordinate metamorphosis with seasonal and light conditions. Melatonin directly antagonized the action of thyroxine (T4) in promoting regression of tadpole tail tips in vitro, and this study sought to determine if it affects the thyroid axis of tadpoles as well. In an experiment sampling at 8-hr intervals for 24 hr, after melatonin treatment (15 micrograms/day for 12 days) of premetamorphic Rana pipiens tadpoles at approximately 1100 hr on 18L:6D, thyroid follicle cell height and lumen diameter were lowered by melatonin, but follicle cell proliferation was not significantly depressed. In a second experiment conducted under the same conditions, but sampling at 3-hr intervals for 24 hr, melatonin significantly lowered follicle cell labeling index and suppressed its ultradian (7.6 hr) rhythm, while shifting the peak of follicle lumen diameter to the dark instead of the light. Thus, melatonin tended to depress the thyroid of young tadpoles and suppress or shift its rhythms. Melatonin (10 micrograms/day for 5 days) injected into prometamorphic Rana catesbeiana tadpoles at 1930 hr on 18L:6D significantly altered subsequent in vitro thyroid function as determined by radioimmunoassay of media collected at intervals for 54 hr from cultured thyroids of injected control and melatonin groups, and a noninjected control group. Melatonin decreased T4 secretion during the first 30 hr, but not during the last 24 hr of culture, suppressed 3,5,3'-triidothyronine (T3) secretion for 12 hr, and then raised T3 output into the media above the control for the remainder of the culture period, increasing the T3:T4 ratio. Injection alone increased both T3 and T4 secretion for the first 30 hr, but did not change the T3:T4 ratio. The findings show that exogenous melatonin administered in vivo significantly modulated thyroid activity and morphometry directly and/or indirectly and comprise the first demonstration of an effect of melatonin on the amphibian thyroid gland.

Circadian rhythms of thyroid secretion, morphometry, and cell division in prometamorphic and climax Rana tadpoles.

Rhythmicity of thyroid follicle structure, cell division, and T4 secretion in vitro was studied in anuran tadpoles on a 12L:12D cycle with light onset at 0800 hr. Stage XIII Rana pipiens larvae had a significant circadian rhythm of follicle lumen diameter (acrophase 1052 hr). Follicle cell height was higher at 0900 hr than at other times but the rhythm was not significant. At Stage XX, both follicle cell height (acrophase 2318 hr) and cell division (acrophase 1929 hr) were rhythmic while lumen diameter, though highest at 0900 hr, showed no significant daily changes. Thyroids from Rana catesbeiana larvae at Stages XVII to XVIII had a significant circadian rhythm of T4 secretion (acrophase 2034 hr) in vitro as measured by radioimmunoassay of media collected every 6 hr for 24 hr. The findings indicate that the daily sequence of thyroid structure and function in preclimax Rana tadpoles includes larger follicle lumina early in the light and maximum cell division, T4 secretion, and follicle cell height within a 4-hr interval beginning around the onset of dark.