Frequent detection of Saffold cardiovirus in adenoids.

Saffold virus (SAFV) is classified into the Cardiovirus genus of the Picornaviridae family. Up to now, eleven genotypes have been identified however, their clinical significance remains unclear. Here, we investigated the presence of SAFV in asymptomatic patients admitted for adenoidectomy. A total of 70 adenoid tissue samples were collected from children with clinical symptoms caused by hypertrophy of adenoids but without symptoms of airway infection. Samples were investigated for SAFV by RT-nested PCR and sequence analysis. Eleven of 70 (15.7%) samples were positive for SAFV. Nasopharyngeal swabs were available from 45 children just before surgery. SAFV was rarely found and only in children with SAFV-positive adenoids 2/8. Our findings indicate that the presence of SAFV seems to be more frequent in adenoid tissue than expected. This could support the notion of a longer than previously anticipated persistence of SAFV nucleic acids in the respiratory tract and possibly a chronic infection. Further investigations are necessary to establish the role of SAFV infection in humans.

Multiplex detection in tonsillar tissue of all known human polyomaviruses.

BACKGROUND: In the past few years, eleven new human viruses have joined the two previously known members JCPyV and BKPyV of the Polyomaviridae family, by virtue of molecular methods. Serology data suggest that infections with human polyomaviruses (HPyVs) occur since childhood and the viruses are widespread in the general population. However, the viral persistence sites and transmission routes are by and large unknown. Our previous studies demonstrated that the four new HPyVs - KIPyV, WUPyV, MCPyV and TSPyV - were present in the tonsils, and suggested lymphoid tissue as a persistent site of these emerging human viruses. We developed a Luminex-based multiplex assay for simultaneous detection of all 13 HPyVs known, and explored their occurrence in tonsillar tissues of children and adults mostly with tonsillitis or tonsillar hypertrophy. METHODS: We set up and validated a new Luminex-based multiplex assay by using primer pairs and probes targeting the respective HPyV viral protein 1 (VP1) genes. With this assay we tested 78 tonsillar tissues for DNAs of 13 HPyVs. RESULTS: The multiplex assay allowed for simultaneous detection of 13 HPyVs with high analytical sensitivity and specificity, with detection limits of 100-102 copies per microliter, and identified correctly all 13 target sequences with no cross reactions. HPyV DNA altogether was found in 14 (17.9%) of 78 tonsils. The most prevalent HPyVs were HPyV6 (7.7%), TSPyV (3.8%) and WUPyV (3.8%). Mixed infection of two HPyVs occurred in one sample. CONCLUSIONS: The Luminex-based HPyV multiplex assay appears highly suitable for clinical diagnostic purposes and large-scale epidemiological studies. Additional evidence was acquired that the lymphoid system plays a role in HPyV infection and persistence. Thereby, shedding from this site during reactivation might take part in transmission of the newly found HPyVs.

A Pilot Study on the Treatment of Posterior Cheek Enlargement in HIV+ Patients With Botulinum Toxin A.

BACKGROUND: Posterior cheek enlargement in human immunodeficiency virus (HIV)+ individuals can lead to significant cosmetic disfigurement. Both parotid gland and masseter muscle overlie the mandibular ramus, contributing to lower facial contour. However, posterior cheek enlargement has not been well characterized anatomically. There are also limited treatment options. Botulinum toxin is a highly efficacious minimally invasive option for improving the shape of the lower face. OBJECTIVE: A pilot study was undertaken to better characterize posterior cheek enlargement in HIV+ patients and explore treatment with botulinum toxin A. MATERIALS AND METHODS: Five HIV+ patients with posterior cheek enlargement were treated with botulinum toxin A. Clinical, photographic, and radiological evaluations allowed the precise calculation of any change in volumes resulting from botulinum toxin A. RESULTS: All 5 patients had a good response with a mean decrease of 21.4% and 11.2% in the volumes of the masseter muscle and parotid gland, respectively. The effect was long lasting even at 6 months after injection and well tolerated. CONCLUSION: Botulinum toxin A may be a less invasive treatment of posterior cheek enlargement in HIV+ patients, with advantages of a good result that is long lasting with good tolerability and minimal risk.

Hypertrophic adenoid is a major infection site of human bocavirus 1.

Human bocavirus 1 (HBoV1) is associated with respiratory infections worldwide, mainly in children. Similar to other parvoviruses, it is believed that HBoV1 can persist for long periods of time in humans, probably through maintaining concatemers of the virus single-stranded DNA genome in the nuclei of infected cells. Recently, HBoV-1 was detected in high rates in adenoid and palatine tonsils samples from patients with chronic adenotonsillar diseases, but nothing is known about the virus replication levels in those tissues. A 3-year prospective hospital-based study was conducted to detect and quantify HBoV1 DNA and mRNAs in samples of the adenoids (AD), palatine tonsils (PT), nasopharyngeal secretions (NPS), and peripheral blood (PB) from patients undergoing tonsillectomy for tonsillar hypertrophy or recurrent tonsillitis. HBoV1 was detected in 25.3% of the AD samples, while the rates of detection in the PT, NPS, and PB samples were 7.2%, 10.5%, and 1.7%, respectively. The viral loads were higher in AD samples, and 27.3% of the patients with HBoV had mRNA detectable in this tissue. High viral loads and detectable mRNA in the AD were associated with HBoV1 detection in the other sample sites. The adenoids are an important site of HBoV1 replication and persistence in children with tonsillar hypertrophy. The adenoids contain high HBoV1 loads and are frequently positive for HBoV mRNA, and this is associated with the detection of HBoV1 in secretions.

Underlying liver disease influences volumetric changes in the spared hemiliver after selective internal radiation therapy with 90Y in patients with hepatocellular carcinoma.

OBJECTIVE: Hypertrophy of the contralateral liver lobe after treatment with yttrium-90 ((90)Y) microspheres has recently been reported. This study aimed to quantify left hepatic lobe hypertrophy after right-sided radioembolization for hepatocellular carcinoma (HCC) and to identify pretreatment predictive factors of hypertrophy in an Asian population. METHODS: A retrospective review of patients with inoperable HCC undergoing selective internal radiation treatment (SIRT) with (90)Y microspheres at a single institution from January 2008 to January 2012 was performed. Only patients who had treatment delivered via the right hepatic artery alone were included. RESULTS: In all, 17 patients fulfilling the study criteria were identified. The mean percentage of left-lobe hypertrophy was 34.2% ± 34.9% (range 19.0-106.5%) during a median of 5-month follow-up. Patients with hepatitis B were found to experience a significantly greater degree of hypertrophy than those with hepatitis C or alcoholic liver cirrhosis. There were no cases of acute liver failure after the administration of SIRT in this study and none of the patients developed disease in the contralateral lobe over the study period. CONCLUSIONS: Administration of unilobar SIRT to the right liver lobe in patients with HCC resulted in a significant degree of contralateral left lobe hypertrophy. Patients with hepatitis B experienced a greater degree of hypertrophy than those with hepatitis C or alcoholic liver cirrhosis.

Prevalence of human papillomavirus and Epstein-Barr virus DNA in Chinese children with tonsillar and/or adenoidal hypertrophy.

Tonsillar and adenoidal hypertrophy are prevalent otolaryngologic disorders in children, but their pathogenesis is largely unknown. The presence of human papillomavirus (HPV) and Epstein-Barr virus (EBV) DNA in 146 tonsil and/or adenoid tissue specimens from 104 Chinese children with tonsillar and/or adenoidal hypertrophy were screened using flow-through hybridization gene-chip technology and real-time fluorescence-based quantitative PCR. Then, the relationships between the prevalence of the viruses and other clinical characteristics of tonsillar and/or adenoidal hypertrophy were analyzed. No patient had HPV DNA. EBV DNA was detected in 19/42 (45.2%) tonsil tissues and 72/104 (69.2%) adenoid tissue specimens (P < 0.05). EBV DNA was not related to the patients' age, gender, disease course, or nationality, but children positive for EBV were less likely to snore; 14/15 (93.3%) patients who did not snore and 59/89 (66.3%) patients who snored were EBV positive. EBV DNA, but not HPV DNA was detected in Chinese children with tonsillar and/or adenoidal hypertrophy. Adenoid tissues might more susceptible than tonsil tissues to EBV infection. In addition, EBV infection did not aggravate snoring in patients with tonsillar and/or adenoidal hypertrophy.

Microbiological profile of adenoid hypertrophy correlates to clinical diagnosis in children.

OBJECTIVE: Adenoid hypertrophy is a common condition in childhood, which may be associated with recurring acute otitis media (RAOM), otitis media with effusion (OME), and obstructive sleep apnea syndrome (OSAS). These different clinical characteristics have some clinical overlap; however, they might be explained by distinct immunologic and infectious profiles and result in various histopathologic findings of adenoid specimens. METHODS: A total of 59 children with adenoid hypertrophy undergoing adenoidectomy were studied. Three series of identical adenoid specimens were processed to hematoxylin-eosin (H.E.) and Gram staining and to respiratory virus specific real-time PCR, respectively. RESULTS: According to the clinical characteristics, patients were recruited into three groups: RAOM (n = 25), OME (n = 19), and OSAS (n = 15). Bacterial biofilms were detected in 21 cases, while at least one of the studied respiratory viruses was detected in 52 specimens. RAOM cases were significantly associated with biofilm existence (n = 20, P < 0.001). In contrast, OME group was characterized by the absence of bacterial biofilm and by normal mucosa. Showing a statistically significant correlation, all OME cases were positive for human bocavirus (HBoV, P < 0.001). CONCLUSIONS: Bacterial biofilms might contribute to the damage of respiratory epithelium and recurring acute infections resulting in RAOM. In OME cases persisting respiratory viruses, mainly HBoV, can cause subsequent lymphoid hyperplasia leading to ventilation disorders and impaired immunoreactivity of the middle ear cleft.

Correlation between structure, protein composition, morphogenesis and cytopathology of Glossina pallidipes salivary gland hypertrophy virus.

The Glossina pallidipes salivary gland hypertrophy virus (GpSGHV) is a dsDNA virus with rod-shaped, enveloped virions. Its 190 kb genome contains 160 putative protein-coding ORFs. Here, the structural components, protein composition and associated aspects of GpSGHV morphogenesis and cytopathology were investigated. Four morphologically distinct structures: the nucleocapsid, tegument, envelope and helical surface projections, were observed in purified GpSGHV virions by electron microscopy. Nucleocapsids were present in virogenic stroma within the nuclei of infected salivary gland cells, whereas enveloped virions were located in the cytoplasm. The cytoplasm of infected cells appeared disordered and the plasma membranes disintegrated. Treatment of virions with 1 % NP-40 efficiently partitioned the virions into envelope and nucleocapsid fractions. The fractions were separated by SDS-PAGE followed by in-gel trypsin digestion and analysis of the tryptic peptides by liquid chromatography coupled to electrospray and tandem mass spectrometry. Using the MaxQuant program with Andromeda as a database search engine, a total of 45 viral proteins were identified. Of these, ten and 15 were associated with the envelope and the nucleocapsid fractions, respectively, whilst 20 were detected in both fractions, most likely representing tegument proteins. In addition, 51 host-derived proteins were identified in the proteome of the virus particle, 13 of which were verified to be incorporated into the mature virion using a proteinase K protection assay. This study provides important information about GpSGHV biology and suggests options for the development of future anti-GpSGHV strategies by interfering with virus-host interactions.

Presence of herpesviruses in adenoid tissues of children with adenoid hypertrophy and chronic adenoiditis.

OBJECTIVES: The aim of study was to determine the presence of some of the herpesviruses including herpes simplex virus (HSV), Epstein-Barr virus (EBV), and cytomegalovirus (CMV) in adenoid tissues of children with adenoid hypertrophy (AH) and chronic adenoiditis (CA) and to investigate the potential role of the herpesviruses in patogenesis of AH and CA. PATIENTS AND METHODS: A total of 72 patients (41 boys, 31 girls; mean age 4 years and 2 months; range 2 to 9 years) who underwent adenoidectomy or adenotonsillectomy (with or without placement of a ventilation tube) in our clinic between October 2007 and May 2008, were included. The patients were divided into two groups, as AH group (n=42) and the CA group (n=30). Adenoid tissues collected from patients in both groups were analyzed by polymerase chain reaction (PCR) for the presence of HSV, EBV and CMV-DNA. RESULTS: The results of the PCR indicated that 33.3% in the AH group and 36.6% in the CA group were herpesvirus DNA positive. Among the herpesviruses studied, HSV-DNA was detected at the highest level (14.2% and 16.6%, respectively) in both groups, although the difference between the groups was not significant. EBV-DNA positiveness was 11.9% and CMV-DNA was 4.7% in the AH group, whereas, EBV-DNA positiveness was 13.3% and CMV-DNA was 6.6% in the CA group. CONCLUSION: Herpesviruses were determined at a high rate in adenoid tissue of children with AH and CA, suggesting that there may be a potential relationship between the presence of herpesviruses and occurrence of AH and CA in children. However, more extensive studies are required to elucidate the role of herpesviruses in the pathogenesis of AH or CA.

HHV-6 infection of tonsils and adenoids in children with hypertrophy and upper airway recurrent infections.

OBJECTIVE: Human herpes virus 6 (HHV-6), the agent of a self-limiting exanthematic disease in childhood, persists in a silent state in the secondary lymphoid organs and the reactivation is characterized by HHV-6-induced inflammatory cytokines. This study investigates the possible etiological role of HHV-6 in children affected by tonsil and adenoid hypertrophy. METHODS: 55 tonsils, 80 adenoids fresh tissues and 74 blood samples were collected from 80 children (mean age 4.8 years, 43.5% female) undergoing elective tonsillectomy and/or adenoidectomy for tissue hypertrophy. Moreover, patients with <5 years old documented upper airway recurrent infections not related to relapsing of acute tonsillitis. Specific IgG antibodies and virus detection (by PCR, variant A/B enzymatic genotyping and real-time PCR) were performed. RESULTS: In our series, HHV-6 seroprevalence was tested at 50%. HHV-6 variant B was the unique strain finding in 25% of adenoids, in 12.7% of tonsils and in 4% of peripheral blood mononuclear cells (PBMCs). HHV-6-B was prevalent in tonsils of children affected by upper airway infections (17.8% vs 7.4%) while the adenoids represented the more frequent reservoir (30.7% vs 19.5%) in patients with hypertrophy. HHV-6 viral load was low, ranging from 80 to 600 copies/10(6) cells suggesting a latent/persistent phase of infection. CONCLUSION: These results reinforce the role of the secondary lymphoid organs as an important reservoir for HHV-6B. Nevertheless, infection of lymphoid cells, sustained by a low level of replication, could be sufficient to increase the local injury through an autologous mechanism of inflammation.

[Study and analysis on the quantitive detection of EBV-DNA in adenoidal hypertrophic and tonsillitis tissues of children].

OBJECTIVE: To investigate the epidemiology of EBV in adenoidal hypertrophy and chronic tonsillitis and discuss the affection of EBV on the nosogenesis of adenoidal hypertrophy and tonsillitis of children. METHOD: Fifty-two children with chronic tonsillitis and/or adenoidal hypertrophy had the operations of the tonsillectomy and/or the adenoidectomy. These tissues resected and plasma of all cases were detected to find EBV-DNA by RQ PCR. RESULT: The infection rate of EBV in the tissues of adenoidal hypertrophy and tonsillitis of children was 51.9%. The boys' infection rate of EBV was 50.0%, and the girls' infection rate of EBV was 55.6%, which had not significantly different. The EBV infection rate in the tissues of tonsillitis was 40.4%, The EBV infection rate in the tissues of adenoidal hypertrophy was 48.9%, which had not significant difference. The school age group (7- to 14-years-old) presented higher infection rate of EBV in the tissues of adenoid and tonsil (65.5%) than the pre-school children group (2- to 6-years-old) (34.8%). Comparing the copies numbers of EBV-DNA in the different degrees of adenoidal hypertrophy, we found that the copies numbers of EBV-DNA in the severe hypertrophy group were higher than the midrange and slight hypertrophy groups (P<0.05). Meanwhile we detected EBV-DNA in these childrens' blood plasma by RQ-PCR. No blood plasma was detected EBV-DNA copies higher than normal (< 1 x 10(3) copies/ml). CONCLUSION: The tissues of adenoidal hypertrophy and tonsillitis had same sensitivity to EBV. There was not significant difference between the infection rates of the boys and girls with adenoidal hypertrophy and/or tonsillitis. With these children growing up and the course of diseases prolonging, the infection rate of EBV increased correspondingly. There was a certain correlation between the hypertrophy of adenoid and EBV. There were no EBV-DNA fragments in blood plasma of the children with adenoidal hypertrophy and/or tonsillitis. So there were essential different between benign hyperplasia and nasopharyngeal carcinoma.