Abstract 20433: Viral Myocarditis: Viral Genome Detection Yield on Endomyocardial Biopsy is Influenced by Methodological Factors

Circulation ◽  
2014 ◽  
Vol 130 (suppl_2) ◽  
Author(s):  
Elisa Carturan ◽  
Stefania Rizzo ◽  
Gaetano Thiene ◽  
Cristina Basso
Keyword(s):  
Endomyocardial Biopsy ◽  
Viral Genome ◽  
Viral Myocarditis ◽  
Molecular Techniques ◽  
Time Interval ◽  
Tissue Fixation ◽  
Viral Genomes ◽  
Consensus Document ◽  
Significant Difference

Introduction: Myocarditis is diagnosed on endomyocardial biopsy (EMB) by histological, immune-histochemical and molecular techniques to detect viral genomes. The role of EMB for the diagnosis and its contribution to patient management has been addressed by a Consensus document of American and European Cardiovascular Pathology Societies. Hypothesis: Different methods of tissue fixation and the number of myocardial samples could impact on viral genome yield on EMB. Methods: Consecutive EMBs referred at our Institution for histology/ immunohistochemistry evaluation in the time interval 1996-2012 were enrolled. Molecular investigation by RT-PCR and PCR technique was performed in all EMBs with inflammatory cardiomyopathy diagnosis at histology and immunohistochemistry. Results: A total of 467 EMBs have been diagnosed as myocarditis: 79 in pediatric (1 month to 18 years; 47M/ 32F) and 388 adult (19 to 75 years; 256M/132 F) patients (pts.). Viral etiology was identified in 28 (36%) pediatric and 101 (26%) adult pts, and the most prevalent type of viruses were enterovirus (8/28, 36%; 26/101, 24%). In a more recent subgroup of 137 EMBs (virus positive 30/137, 22%) seasonality, type of fixation and number of EMB samples were assessed. The seasonal distribution of myocarditis was higher in winter than in other seasons (33% vs. autumn 28%, spring 20% and summer 19%) without any significant difference in terms of virus positive EMBs. The number of EMB samples per pt. was ≤3 (either formalin or RNAlater) in 94/137 (69%) and >3 in 43/137 (31%), with a lower prevalence of virus positive in the former (17/94, 18% vs. 13/43, 30%). Eighty-one EMB samples were frozen while 56 were paraffin embedded, with a higher prevalence of viral genome in the former (26/81, 32% vs. 4/56, 7%; p=0.001). Conclusions: The diagnosis of myocarditis on EMB samples requires standardized protocols including molecular techniques. Viral genomes are identifiable in more than one third of pediatric and one fourth of adult cases. Methodological factors like the type of tissue fixation and the number of samples could impact on viral genome detection on EMB.

scholarly journals Molecular detection of cattle Sarcocystis spp. in North-West Italy highlights their association with bovine eosinophilic myositis

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Selene Rubiola ◽  
Tiziana Civera ◽  
Felice Panebianco ◽  
Davide Vercellino ◽  
Francesco Chiesa
Keyword(s):  
18S Rdna ◽  
Inflammatory Myopathy ◽  
Molecular Techniques ◽  
Diaphragm Muscle ◽  
Economic Losses ◽  
Intermediate Hosts ◽  
Slaughter Cattle ◽  
Significant Difference ◽  
Pcr Targeting

Abstract Background Cattle are intermediate hosts of six Sarcocystis species, among which Sarcocystis hominis and Sarcocystis heydorni can infect humans through the consumption of raw or undercooked meat. In addition to the zoonotic potential, there is increasing interest in these protozoa because of the evidence supporting the role of Sarcocystis spp. in the occurrence of bovine eosinophilic myositis (BEM), a specific inflammatory myopathy which leads to carcass condemnation and considerable economic losses. Actually, all the prevalence studies carried out on cattle in Italy have been based on either morphological or 18S rDNA-based molecular techniques, most likely leading to misidentification of closely related species. Therefore, there is a strong need for new data on the prevalence of the different Sarcocystis spp. in cattle in Italy and their association with bovine eosinophilic myositis. Methods To reach our aim, individual striated muscle samples from BEM condemned carcasses (N = 54) and diaphragm muscle samples from randomly sampled carcasses (N = 59) were obtained from Northwest Italy slaughterhouses. Genomic DNA was extracted and analyzed by multiplex-PCR targeting 18S rDNA and cox1 genes. PCR products amplified using the genus-specific primer set in absence of the specific fragment for S. hirsuta, S. cruzi, S. hominis or S. bovifelis were sequenced to achieve species identification. Results Sarcocystis DNA was detected in 67.8% of the samples from slaughter cattle and in 90.7% of the samples from BEM condemned carcasses. S. cruzi was identified as the most prevalent species in slaughter cattle (61%), followed by S. bovifelis (10.2%), S. hominis (8.5%) and S. hirsuta (1.7%). Notably, among the different Sarcocystis spp. detected, the presence of S. bovifelis and S. hominis was significantly higher in samples isolated from BEM condemned carcasses (46.3% and 40.7% respectively), while there was no statistically significant difference between the presence of S. cruzi or S. hirsuta in BEM condemned carcasses (42.6% and 1.8%, respectively) and randomly sampled carcasses. Furthermore, DNA sequence analysis revealed the presence of a putative new species in two carcasses. Conclusions Our study contributes to updating the data on the prevalence of the different Sarcocystis spp. in cattle in Italy, highlighting the presence of three Sarcocystis spp., S. cruzi, S. hominis and S. bovifelis, in BEM lesions and allowing us to speculate on the possible role of S. hominis and S. bovifelis as the major sarcosporidian species involved in bovine eosinophilic myositis. Graphic Abstract

Inheritance of Chromosomally Integrated Human Herpesvirus 6 DNA

Blood ◽  
1999 ◽  
Vol 94 (5) ◽  
pp. 1545-1549 ◽  
Author(s):  
Masanori Daibata ◽  
Takahiro Taguchi ◽  
Yuiko Nemoto ◽  
Hirokuni Taguchi ◽  
Isao Miyoshi
Keyword(s):  
Dna Sequences ◽  
Viral Genome ◽  
Family Members ◽  
Human Herpesvirus ◽  
Single Step ◽  
Human Herpesvirus 6 ◽  
Viral Genomes ◽  
Herpesvirus 6

Abstract Human herpesvirus 6 (HHV-6) genome has been detected in several human lymphoproliferative disorders with no signs of active viral infection, and found to be integrated into chromosomes in some cases. We previously reported a woman with HHV-6–infected Burkitt’s lymphoma. Fluorescence in situ hybridization showed that the viral genome was integrated into the long arm of chromosome 22 (22q13). The patient’s asymptomatic husband also carried HHV-6 DNA integrated at chromosome locus 1q44. To assess the possibility of chromosomal transmission of HHV-6 DNA, we looked for HHV-6 DNA in the peripheral blood of their daughter. She had HHV-6 DNA on both chromosomes 22q13 and 1q44, identical to the site of viral integration of her mother and father, respectively. The findings suggested that her viral genomes were inherited chromosomally from both parents. The 3 family members were all seropositive for HHV-6, but showed no serological signs of active infection. To confirm the presence of HHV-6 DNA sequences, we performed polymerase chain reaction (PCR) with 7 distinct primer pairs that target different regions of HHV-6. The viral sequences were consistently detected by single-step PCR in all 3 family members. We propose a novel latent form for HHV-6, in which integrated viral genome can be chromosomally transmitted. The possible role of the chromosomally integrated HHV-6 in the pathogenesis of lymphoproliferative diseases remains to be explained.

scholarly journals Italian observational study on HPV infection, E6, and p16 expression in men with penile cancer

2020 ◽  
Vol 17 (1) ◽  
Author(s):  
Narcisa Muresu ◽  
Giovanni Sotgiu ◽  
Laura Saderi ◽  
Illari Sechi ◽  
Antonio Cossu ◽  
...  
Keyword(s):  
Penile Cancer ◽  
Tertiary Care ◽  
Hpv Infection ◽  
Molecular Techniques ◽  
Care Hospital ◽  
Hpv 16 ◽  
Hpv Dna ◽  
Diagnosis And Prognosis ◽  
Significant Difference

Abstract Background Human Papillomavirus (HPV) infection is one of the most important causes of cancer. It can play a role in cervical and extra-cervical cancers. Penile cancer is rare, even if an increasing trend was recently reported. Aim of the present study was to assess the prevalence and distribution of HPV genotypes in cases of penile cancer diagnosed in Sardinia, Italy. Surrogate markers of HPV infection (i.e., E6 and p16 genes) were also evaluated in all cases. Methods An observational, retrospective study which recruited all cases of penile cancer diagnosed between 2002 and 2019 at a tertiary care hospital in Sardinia, Italy, was carried out. HPV-DNA detection and genotyping were performed by Real-time PCR. Specimens were tested for oncogene E6 mRNA and for p16(INK4a) expression. Results HPV prevalence was 28.1% (9/32); HPV-16 was the most prevalent genotype (7/9, 77.8%). p16INK4a positivity was found in 66.7% of the samples with a statistically significant difference between HPV-positive and -negative groups. E6-transcript was detected in 71% of the HPV-16 positive samples. The overall survival was not statistically different between HPV-positives and -negatives. Discussion The present study confirms the etiologic role of HPV in penile cancer and supports the adoption of vaccination strategies in men and women. Further studies should clarify the diagnostic and prognostic role of E6 and p16 proteins. Conclusion HPV infection can favor the occurrence of penile cancer, whose diagnosis and prognosis could be improved with the implementation of validated molecular techniques.

Inheritance of Chromosomally Integrated Human Herpesvirus 6 DNA

Blood ◽  
1999 ◽  
Vol 94 (5) ◽  
pp. 1545-1549 ◽  
Author(s):  
Masanori Daibata ◽  
Takahiro Taguchi ◽  
Yuiko Nemoto ◽  
Hirokuni Taguchi ◽  
Isao Miyoshi
Keyword(s):  
Dna Sequences ◽  
Viral Genome ◽  
Family Members ◽  
Human Herpesvirus ◽  
Single Step ◽  
Human Herpesvirus 6 ◽  
Viral Genomes ◽  
Herpesvirus 6

Human herpesvirus 6 (HHV-6) genome has been detected in several human lymphoproliferative disorders with no signs of active viral infection, and found to be integrated into chromosomes in some cases. We previously reported a woman with HHV-6–infected Burkitt’s lymphoma. Fluorescence in situ hybridization showed that the viral genome was integrated into the long arm of chromosome 22 (22q13). The patient’s asymptomatic husband also carried HHV-6 DNA integrated at chromosome locus 1q44. To assess the possibility of chromosomal transmission of HHV-6 DNA, we looked for HHV-6 DNA in the peripheral blood of their daughter. She had HHV-6 DNA on both chromosomes 22q13 and 1q44, identical to the site of viral integration of her mother and father, respectively. The findings suggested that her viral genomes were inherited chromosomally from both parents. The 3 family members were all seropositive for HHV-6, but showed no serological signs of active infection. To confirm the presence of HHV-6 DNA sequences, we performed polymerase chain reaction (PCR) with 7 distinct primer pairs that target different regions of HHV-6. The viral sequences were consistently detected by single-step PCR in all 3 family members. We propose a novel latent form for HHV-6, in which integrated viral genome can be chromosomally transmitted. The possible role of the chromosomally integrated HHV-6 in the pathogenesis of lymphoproliferative diseases remains to be explained.

scholarly journals Molecular Detection of Cattle Sarcocystis Spp. In North-West Italy Targeting Cox1 And 18S Genes Highlights Its Association With Bovine Eosinophilic Myositis.

2021 ◽  
Author(s):  
Selene Rubiola ◽  
Tiziana Civera ◽  
Felice Panebianco ◽  
Davide Vercellino ◽  
Francesco Chiesa
Keyword(s):  
Inflammatory Myopathy ◽  
Molecular Techniques ◽  
Economic Losses ◽  
Intermediate Hosts ◽  
Prevalence Studies ◽  
North West ◽  
Slaughter Cattle ◽  
Significant Difference ◽  
Pcr Targeting

Abstract Background: Cattle are intermediate hosts of six Sarcocystis species, among which Sarcocystis hominis and Sarcocystis heydorni can infect humans through the consumption of raw or undercooked meat. In addition to the zoonotic potential, there is increasing interest in these protozoa due to the evidences supporting the role of Sarcocystis spp. in the occurrence of bovine eosinophilic myositis (BEM), a specific inflammatory myopathy which leads to carcass condemnation and considerable economic losses. Actually, all the prevalence studies carried out on cattle in Italy have been based either on morphological or 18S rDNA-based molecular techniques, most likely leading to misidentification of closely related species. Therefore, there is a strong need for new data on the prevalence of the different Sarcocystis spp. in cattle in Italy and their association with bovine eosinophilic myositis.Methods: To reach our aim, individual cattle samples from BEM condemned carcasses (N=54) and randomly sampled carcasses (N=59) were obtained from Piedmont slaughterhouses. Genomic DNA was extracted and analyzed by multiplex-PCR targeting 18S and cox1 genes. PCR products amplified using the genus specific primer set in absence of the specific fragment for S. hirsuta, S. cruzi, S. hominis or S. bovifelis, were sequenced to achieve species identification.Results: Sarcocystis DNA was detected in 67.8% of the samples from slaughter cattle and in 90.7% of the samples from BEM condemned carcasses. S. cruzi was identified as the most prevalent species (61%), followed by S. bovifelis (10.2%), S. hominis (8.5%) and S. hirsuta (1.7%). Notably, among the different Sarcocystis spp. detected, the presence of S. bovifelis and S. hominis was significantly higher in samples isolated from BEM condemned carcasses (46.3% and 40.7% respectively), while there was no statistically significant difference between the presence of S. cruzi or S. hirsuta in BEM condemned carcasses and randomly sampled carcasses. Furthermore, DNA sequence analysis revealed the presence of a putative new species in 2 carcasses.Conclusions: Our study contributes to update the data on the prevalence of the different Sarcocystis spp. in cattle in Italy and emphasize the role of S. hominis and S. bovifelis as the major sarcosporidian species involved.

scholarly journals Uncovering the Role of the E1 Protein in Different Stages of Human Papillomavirus 18 Genome Replication

2020 ◽  
Vol 94 (20) ◽  
Author(s):  
Alla Piirsoo ◽  
Martin Kala ◽  
Eve Sankovski ◽  
Mart Ustav ◽  
Marko Piirsoo
Keyword(s):  
Life Cycle ◽  
Viral Genome ◽  
Copy Number ◽  
Genome Replication ◽  
Dependent Manner ◽  
Infection Cycle ◽  
Viral Genomes ◽  
E1 Protein ◽  
Viral Genome Replication

ABSTRACT The life cycle of human papillomaviruses (HPVs) comprises three distinct phases of DNA replication: initial amplification, maintenance of the genome copy number at a constant level, and vegetative amplification. The viral helicase E1 is one of the factors required for the initiation of HPV genome replication. However, the functions of the E1 protein during other phases of the viral life cycle are largely uncharacterized. Here, we studied the role of the HPV18 E1 helicase in three phases of viral genome replication by downregulating E1 expression using RNA interference or inducing degradation of the E1 protein via inhibition of casein kinase 2α expression or catalytic activity. We generated a novel modified HPV18 genome expressing Nanoluc and tagged E1 and E2 proteins and created several stable HPV18-positive cell lines. We showed that, in contrast to initial amplification of the HPV18 genome, other phases of viral genome replication involve also an E1-independent mechanism. We characterize two distinct populations of HPV18 replicons existing during the maintenance and vegetative amplification phases. We show that a subset of these replicons, including viral genome monomers, replicate in an E1-dependent manner, while some oligomeric forms of the HPV18 genome replicate independently of E1 function. IMPORTANCE Human papillomavirus (HPV) infections pose serious medical problem. To date, there are no HPV-specific antivirals available due to poor understanding of the molecular mechanisms of virus infection cycle. The infection cycle of HPV involves initial amplification of the viral genomes and maintenance of the viral genomes with a constant copy number, followed by another round of viral genome amplification and new viral particle formation. The viral protein E1 is critical for the initial amplification of the viral genome. However, E1 involvement in other phases of the viral life cycle has remained controversial. In the present study, we show that at least two different replication modes of the HPV18 genome are undertaken simultaneously during the maintenance and vegetative amplification phases, i.e., replication of the majority of the HPV18 genome proceeds under the control of the host cell replication machinery without E1 function, whereas a minority of the genome replicates in an E1-dependent manner.

Effect of Dipyridamole on Spontaneous Platelet Aggregation in Whole Blood Decreases with the Time After Venepuncture: Evidence for the Role of ADP

1987 ◽  
Vol 58 (02) ◽  
pp. 744-748 ◽  
Author(s):  
A R Saniabadi ◽  
G D O Lowe ◽  
J C Barbenel ◽  
C D Forbes
Keyword(s):  
Platelet Aggregation ◽  
Correlation Coefficient ◽  
Whole Blood ◽  
Blood Platelet ◽  
Inhibitory Effect ◽  
Time Interval ◽  
Adp Receptor ◽  
Spontaneous Platelet Aggregation

SummarySpontaneous platelet aggregation (SPA) was studied in human whole blood at 3, 5, 10, 20, 30, 40 and 60 minutes after venepuncture. Using a whole blood platelet counter, SPA was quantified by measuring the fall in single platelet count upon rollermixing aliquots of citrated blood at 37° C. The extent of SPA increased with the time after venepuncture, with a correlation coefficient of 0.819. The inhibitory effect of dipyridamole (Dipy) on SPA was studied: (a) 10 μM at each time interval; (b) 0.5-100 μM at 3 and 30 minutes and (c) 15 μM in combination with 100 μM adenosine, 8 μM 2-chloroadenosine (2ClAd, an ADP receptor blocker) and 50 μM aspirin. There was a rapid decrease in the inhibitory effect of Dipy with the time after venepuncture; the correlation coefficient was -0.533. At all the concentrations studied, Dipy was more effective at 3 minutes than at 30 minutes after venepuncture. A combination of Dipy with adenosine, 2ClAd or aspirin was a more effective inhibitor of SPA than either drug alone. However, when 15 μM Dipy and 10 μM Ad were added together, the inhibitory effect of Dipy was not increased significantly, suggesting that Dipy inhibits platelet aggregation independent of Ad. The increase in SPA with the time after venepuncture was abolished when blood was taken directly into the anticoagulant containing 5 μM 2ClAd. It is suggested that ADP released from the red blood cells is responsible for the increased platelet aggregability with the time after venepuncture and makes a serious contribution to the artifacts of in vitro platelet function studies.

Role of Teachers in Building the Concept of Sustainable Development: Success or Failure

2020 ◽  
Vol 1 (1) ◽  
Author(s):  
Ayaz Muhammad Khan ◽  
Amber Jamshaid ◽  
Tayyibah Roohi ◽  
Amna Ramzan
Keyword(s):  
Sustainable Development ◽  
Primary School ◽  
Sampling Technique ◽  
School Teachers ◽  
Primary School Teachers ◽  
Primary Level ◽  
Significant Difference ◽  
Role Of Teachers ◽  
Major Factors

Sustainable Development (SD) is a rich, challenging and thought-provoking construct in social sciences. The main purpose of this paper was to identify and explore the role played by primary school teachers in building up the idea of sustainable development (SD) among students. This paper was intended to identify that how a teacher can successfully execute the concept of SD by influencing students’ minds at the primary level. Quantitative survey technique were utilized for data collection. All the primary school teachers of Lahore division comprised the population of the study. Through multistage sampling technique, 352 primary school teachers were selected as participants of the study. A self-developed SD questionnaire incorporating four major factors (teachers’ awareness, pedagogy, curricular and co-curricular activities) with Cronbach’s alpha value = .93 was used to measure the role of teachers in building the sustainability concept among students at primary level. The results indicated a significant mean score difference among SD scores of teachers, sector wise (private and public). Furthermore, the results also reconnoitered the significant difference (p=.04) between the mean scores of female and male teachers in building up the SD concept in students’ minds.

Effectiveness Of Toilet Training Education On Mother's Behavior And Toddler Age Toilet Training Ability (18-36 Months)

2020 ◽  
Vol 5 (2) ◽  
pp. 49-55
Author(s):  
Hafiko Andresni ◽  
Zahtamal Zahtamal ◽  
Winda Septiani ◽  
Mitra Mitra ◽  
Lita Lita
Keyword(s):  
Sampling Technique ◽  
T Test ◽  
The Body ◽  
Toilet Training ◽  
Wilcoxon Test ◽  
Control Group ◽  
Maternal Knowledge ◽  
Control Group Design ◽  
Significant Difference

ABSTRACT Toilet training is an effort to train children to be able to control and urinate (BAK) and defecate (BAB). Toilet training is one of the main tasks of children at toddler age. Toilet training is one of the main tasks of children in toddler age which is very important to be done to create independence in children in controlling BAK and BAB and children know the parts of the body and their functions. Data in 2012 shows that ± 60% of parents do not teach toilet training to children from an early age. The aim of the study was to find out the effectiveness of toilet training education on maternal behavior and toilet skills in toddler age training (18-36 months). The study was conducted in July-August 2018. This type of quantitative research used the design of the Quasy pretest and posttest experiment with non-equivalent control group design. Samples were 36 mothers and 36 children with purposive sampling technique. Data analysis used Paired t test, Wilcoxon test, Man-Whitney test an Independent t test. The results showed that toilet training education through lecture methods, modules and maze games was more effective than toilet training education through lecture and leaflet methods on children's knowledge and abilities. Conversely, for the role of mothers in supervision there is no significant difference in effectiveness. Health education is recommended in health promotion programs to increase maternal knowledge, the role of mothers and the ability of toilet training children independently. Keywords: Toilet training, Lecture method, Module, Maze game, Leaflet, Knowledge, Role of mother, Children's ability.

SUBLINGUAL VITAMIN D3 DRUG THERAPY IN VITAMIN D DEFICIENCY PATIENTS AT PRAVARA RURAL HOSPITAL

2019 ◽  
pp. 18-20
Author(s):  
Sanjeeva Kumar Goud T ◽  
Rahul Kunkulol
Keyword(s):  
Vitamin D ◽  
Vitamin D Deficiency ◽  
Vitamin D3 ◽  
Serum Vitamin ◽  
Cross Sectional ◽  
Serum Vitamin D ◽  
Significant Difference ◽  
Before And After

The present study was aimed to study the effect of Sublingual Vitamin D3 on Serum Vitamin D level in Vitamin D deficiency patients. This was a cross-sectional and interventional study. All the Vitamin D deficiency patients of age 18-60years and either gender, willing to participate in the study were included. Patients who had greater than 20 ng/ml were excluded from the study. The total number of participants in our study was 200, out of these 111 males and 89 females, the mean age in our study was 51.07 ± 7.39Yrs. All volunteers were given sublingual vitamin D3 (60,000IU) in six doses every fifteen days of follow up for 3 months. The subject’s serum 25(OH)D levels were estimated before and after the treatment of sublingual vitamin D3. There was a statistically significant difference in serum vitamin D3 level before 16.61±6.71 ng/ml and after 35.80±7.80 ng/ml after treatment with Sublingual Vitamin D3. Six doses of 60,000IU of Vitamin D3 sublingual route having improved the role of serum 25(OH)D levels in the treatment of Vitamin D3 deficiency patients.Keywords: Vitamin D3; Sublingual route

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