The influence of interferon-lambda on restricting Middle East Respiratory Syndrome Coronavirus replication in the respiratory epithelium.

Middle East Respiratory Syndrome Coronavirus (MERS-CoV) causes severe respiratory in human with high mortality and it has been a challenge to determine optimum treatment for MERS-CoV-induced respiratory infection. Here, we observed the distribution of MERS-CoV receptors using human respiratory mucosa and also evaluated the contribution of interferon-lambdas (IFN-λs) in response to MERS-CoV infection using in vitro normal human nasal epithelial (NHNE) and bronchial epithelial (NHBE) cells. We found that the gene and protein expression of DPPIV, MERS-CoV receptor, were more dominantly located in nasal and bronchial epithelium although human nasal mucosa exhibited relatively lower DPPIV expression than lung parenchymal tissues. The quantitative mRNA level of the MERS-CoV envelope (upE) gene was significantly induced in MERS-CoV-infected cultured NHNE and NHBE cells until 3 days after infection. The induction of IFNs was identified in NHNE and NHBE cells after MERS-CoV infection and IFN-λs were predominantly increased in MERS-CoV-infected respiratory epithelial cells. Inoculation of IFN-λs to NHNE and NHBE cells suppressed MERS-CoV replication and in particular, IFN-λ4 showed a strong therapeutic effect in reducing MERS-CoV infection with higher induction of IFN-stimulated genes. Thus, IFN-λ has a decisive function in the respiratory epithelium that greatly limits MERS-CoV replication, and may be a key cytokine for better therapeutic outcomes against MERS-CoV infection in respiratory tract.

COVID-19 Pandemic and its Impact on Craniofacial Surgery.

In late 2019, a novel coronavirus strain, SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2), also known as coronavirus disease 2019 (COVID-19), triggered a global pandemic as the virus spread from the Wuhan Province, China, across all continents. Although infrequent, severe respiratory infection and death caused by SARS-CoV-2 is disproportionately high amongst healthcare providers such as craniofacial surgeons who work in the head and neck region. Factors this impact SARS-CoV-2 transmission include: (1) high viral loads in the mucosa of the oral and nasopharynx, (2) limited and/or imprecise disease screening/confirmation testing, (3) access to and appropriate use of personal protective equipment (PPE).

Herpes zoster laryngitis with intractable hiccups.

A 73-year-old man presented to our hospital with a sore throat (left-sided) and hiccups. The patient had mucosal swelling and erosions affecting the left posterior pillar, base of tongue, epiglottis, arytenoid, and aryepiglottic fold. As the laryngeal mucosal edema became worse, herpetic vesicles and erosions developed on the left cavum conchae, external auditory canal, and palate. The patient was treated with acyclovir and a steroid. His hiccups were treated with metoclopramide, but it had little effect, and hiccups only subsided gradually after the disappearance of erosions. His hiccups relapsed transiently with vomiting, and then resolved completely. Elevation of the CF titer after 2 weeks confirmed the diagnosis of herpes zoster. This condition should be considered in patients with unilateral sore throat and intractable hiccups, and treatment with acyclovir should be provided.

Lycopene enrichment of cultured airway epithelial cells decreases the inflammation induced by rhinovirus infection and lipopolysaccharide.

Rhinovirus infection results in increased release of inflammatory mediators from airway epithelial cells in asthma. As an antioxidant, lycopene offers protection from adverse effects of inflammation. The aim of this study was to find an appropriate method of lycopene enrichment of airway epithelial cells and to determine the effects of lycopene enrichment on the inflammatory response of cells infected by rhinovirus or exposed to lipopolysaccharide. Lycopene enrichment of airway epithelial cells using solubilisation in tetrahydrofuran versus incorporation in liposomes was compared. After determining that solubilisation of lycopene in tetrahydrofuran was the most suitable method of lycopene supplementation, airway epithelial cells (Calu-3) were incubated with lycopene (dissolved in tetrahydrofuran) for 24 h, followed by rhinovirus infection or lipopolysaccharide exposure for 48 h. The release of interleukin-6, interleukin-8 and interferon-gamma induced protein-10 (IP-10) and their messenger RNA levels were measured using enzyme linked immunosorbent assay and reverse transcription polymerase chain reaction, respectively. Viral replication was measured by tissue culture infective dose of 50% assay. Lycopene concentration of cells and media were analysed using high-performance liquid chromatography. Preincubation of airway epithelial cells with lycopene (dissolved in tetrahydrofuran) delivered lycopene into the cells and resulted in a 24% reduction in interleukin-6 after rhinovirus-1B infection, 31% reduction in IP-10 after rhinovirus-43 infection and 85% reduction in rhinovirus-1B replication. Lycopene also decreased the release of IL-6 and IP-10 following exposure to lipopolysaccharide. We conclude that lycopene has a potential role in suppressing rhinovirus induced airway inflammation.

Intralesional Cidofovir application in recurrent laryngeal papillomatosis.

Recurrent laryngeal papillomatosis is a benign disease of the larynx often leading to organic and functional restrictions. The therapeutic treatment of choice in larynx-obstructing papillomatosis is at present surgical laser ablation. The effectiveness of adjuvant intralesional injection of the virustaticum Cidofovir has been investigated recently in a variety of therapeutic models. The present case study deals with the treatment of recurrent laryngeal papillomatosis by means of surgical laser ablation of the laryngeal papillomas with adjuvant local injection of the virustaticum Cidofovir (dose of 5 mg/1 ml). Within the period from October 2001 to August 2004, ten patients aged between 5- and 70 years were treated with intralesional injections of Cidofovir. Papillomatosis was confirmed histologically in all cases, and the virus types were defined in part. Each of the patients underwent clinical-phoniatric examinations and was photographed for documentation. After 2-7 treatments with surgical laser papilloma ablation and intralesional Cidofovir injections, all patients showed a definite papilloma reduction, while in six cases complete remission was achieved. During the follow-up period of 8-30 months, not a single recurrence of the laryngeal papillomatosis occurred. In the majority of patients, a clear improvement in the voice was achieved. There were no local or systemic side effects caused by the virustaticum. Intralesional injection of Cidofovir is an adjuvant, but not a curative therapeutic option in recurrent laryngeal papillomatosis. Remission of previously frequently recurrent laryngeal papillomas can be achieved, but recurrence after longer treatment-free intervals is also possible.

The prevalence of human papillomavirus in the oropharynx in healthy individuals in a Brazilian population.

Oncogenic human papillomavirus (HPV), a causative agent of uterine cervical cancer, has also been detected in head and neck squamous cell cancers, especially in squamous cell carcinomas of the tonsils. However, the true HPV prevalence in normal and neoplasic oropharyngeal mucosa remains uncertain. To determine the prevalence of HPV DNA in normal oropharyngeal mucosa of cancer-free individuals, a study was carried out on 50 Brazilian subjects. PCR was performed to identify HPV DNA in samples from four sites in the oropharynx (tonsils, soft palate, base of the tongue, and back wall of the pharynx). For amplification of the HPV DNA, MY09/11 consensus primers were used, and specific genotypes were identified by dot-blot hybridization or cloning and sequencing. HPV DNA was present in 14.0% of the individuals, and the identified genotypes were 16, 18, 52, and 61. All these types are considered high-risk (HR) HPV. The tonsils and the soft palate were the sites with the highest HPV prevalence. This study shows the prevalence of HR HPV in the oropharynx of normal individuals. However, the prevalence of HPV is still unclear, and if HPV infection in a healthy it is not known individual predisposes to HPV-associated disease such as oropharyngeal cancer. Thus, it is important to assess the prevalence of HPV in cancer-free individuals, in order to compare it with the HPV prevalence in oropharyngeal carcinomas and to attempt to determine the true role of HPV in the development of head and neck squamous cell cancers.

[The prevalence of E6/E7 HPV type 16 in laryngeal cancer and in normal mucosa]. / Obecnosc E6/E7 wirusa HPV16 w raku krani oraz w makroskopowo niezmienionej blonie sluzowej z marginesu operacynego.

Many authors suggest that HPV infection can play a great role in development of benign and malignant tumours of upper respiratory tract in human. The aim of this study was to determine the prevalence of E6/E7 HPV-16 in laryngeal squamous cell carcinoma, and normal laryngeal mucosa, and to analyse their correlation with sex, lymph node status, primary tumor stage, localization, and histological differentiation. HPV 16 DNA presence was analysed using PCR technique in 72 samples of laryngeal carcinoma and in samples of 50 normal mucosa. Human papillomavirus was detected in 26 (36.1%) of the 72 patients. There was no statistically significant correlation HPV positivity and clinicopathological features of the analysed group. In 5 (10%) of 50 samples of normal mucosal. HPV 16 presence in normal mucosa and in laryngeal squamous cell carcinoma was detected in 3 patients. Our observations suggest that HPV16 can play a role in pathogenesis of laryngeal cancer.

[Papilloma virus infection in pretumor and tumor masses of the larynx]. / Papillomavirusnaia infektsiia v predopukholevykh i opukholevykh obrazovaniiakh gortani.

There has been recent evidence suggesting that papilloma virus infection is of no small importance in the pathogenesis of tumors of the head and neck. This study has involved a screening of the mucosa having laryngeal pretumors and tumors for papilloma virus infection by polymerase chain reaction and immunohistochemistry. Specimens were studied in 130 patients. All the patients with laryngeal papillomatosis were found to have human papilloma virus (HPV) types 6 and 11. HPV-18 was detected in 6 (13%) of 48 cases of laryngeal cancer and in 4 (9%) of 46 cases of laryngeal mucosal precancer. Mixed HPV infection of 2-3 different types was revealed in 6 (20%) of 30 HPV-positive patients. Such a wide survey of laryngeal pretumors and tumors in the Russian population for virus infection has been conducted for the first time.

Dendritic cell influx differs between the subglottic and glottic mucosae during acute laryngotracheitis induced by a broad spectrum of stimuli.

Clinically, the subglottic and glottic mucosae may react differently, eg, during acute laryngotracheitis. In healthy rats, we showed previously that the composition of the mucosal immune system of the larynx also differs between these areas. Neutrophils, lymphocytes, and dendritic cells (DCs) are part of this mucosal immune system. In particular, DCs occupy a key function. They migrate into inflamed mucosae during the early phase of the immune response, which is normally characterized by an influx of neutrophils. Thus, they help to overcome the time lag between the innate and the adaptive immune responses. In the present study, the influx of DCs, neutrophils, and T lymphocytes into the subglottic and glottic mucosae of rats was examined at different time points after challenge with a broad spectrum of stimuli such as dead Moraxella catarrhalis, viable Bordetella pertussis, viable Sendai virus, and the soluble protein ovalbumin. The number of DCs increased rapidly after the application of the antigens. This increase was as rapid as the increase in neutrophils. Depending on the kind of antigen, their number in the mucosa increased up to 1,000 cells per 0.1 mm2 (Sendai virus). The comparison of different mucosal areas shows that an overwhelming number of immunocompetent cells entered the subglottic mucosa, whereas only a few cells migrated into the adjacent glottic mucosa. In conclusion, after inhalation of different kinds of antigens, the subset of immunocompetent cells investigated in this study entered the laryngeal mucosa in high numbers. The number of DCs entering the laryngeal mucosa was higher than the numbers of the other immune cells investigated. This finding underlines their function as first-line sentinels of the mucosal immune system of the larynx. The observation that the number of cells entering the laryngeal mucosa is location-dependent indicates the ability of adjacent laryngeal regions to react differently. This is similar to the clinical observation of a selective subglottic reaction during acute laryngotracheitis.

Cellular manifestations of human papillomavirus infection in laryngeal tissues.

BACKGROUND AND OBJECTIVES: Although epidemiologic studies have suggested human papillomavirus (HPV) to be an etiological agent in laryngeal carcinogenesis, little is known on the cellular manifestations of HPV infection in these tumors. In this study, we investigated the frequency of HPV infection in various neoplastic and non-neoplastic laryngeal tissue and its association with expression of the proliferating cell nuclear antigen (PCNA) and the tumor suppressor protein p53. METHODS: Tissues were analyzed by polymerase chain reaction (PCR) for the presence of HPV and by immunocytochemistry for the expression of p53 and PCNA. RESULTS: None of the six normal laryngeal tissues showed the presence of HPV. Thirteen out of the 16 papillomas were positive for HPV, while 15 out of the 44 invasive cancers were HPV positive. PCNA expression increased as the lesion progressed through increasing histological abnormality (r = 0.64400, P = 0.00000). The correlation between the type of laryngeal neoplasm and p53 accumulation was significant (r = 0.54839, P = 0.00000). Significant correlation was also evident between presence of HPV and p53 accumulation (r = 0.34259, P = 0.00424) and PCNA expression (r = 0.036024, P = 0.00266) indicating that HPV positive tumors showed significant p53 accumulation and increased proliferation. There was also correlation between p53 and PCNA expression (r = 0.67475, P = 0.00000) indicating that in all tumors with p53 accumulation, there was a corresponding increase in PCNA expression. CONCLUSIONS: The results suggests that changes in p53 and PCNA expression may be associated with HPV infection, and could play a role in laryngeal carcinogenesis.

Mucosal swabs detect HPV in laryngeal papillomatosis patients but not family members.

Seven patients, aged 2-7 years, with active recurrent respiratory papillomatosis (RRP) attending the University of Michigan Pediatric Otolaryngology Clinic were studied to determine if human papillomavirus (HPV) is harbored in sites of the upper aerodigestive tract other than in the laryngeal papilloma itself. We also determined if close family members had detectable virus in their oral cavities. Noninvasive swabs of buccal mucosa, posterior pharynx, nasal vestibule, and tonsillar pillar of patients, as well as buccal mucosa and posterior pharyngeal swabs of family members were studied. Swabs of the patients' papillomas served as the positive controls. HPV was detected using polymerase chain reaction (PCR) amplification and Southern hybridization techniques. Six of seven patients had detectable HPV in papilloma and endolaryngeal swabs. Four were HPV type 6, and two were HPV type 11. The patient whose swab was negative for HPV was found to be biopsy negative for papilloma 3 weeks after a single laser excision which was performed 6 months prior to the endolaryngeal swab. HPV types 16, 18 and 31 were not found in any of the patients. No swabs from other sites in patients or family members were HPV positive despite the presence of adequate DNA in the swabbed material for successful amplification of beta-actin sequences. The absence of HPV (other than in the papilloma itself) in the upper aerodigestive tract of patients and caregivers is consistent with the absence of reported cases of horizontal transmission to siblings or other family members. The findings are also consistent with the conventional view that juvenile respiratory HPV is transmitted vertically from vaginal condylomas in the mother.

Physical state and expression of human papillomavirus in laryngeal carcinoma and surrounding normal mucosa.

Epidemiologic and biomolecular evidence suggests that human papillomavirus (HPV) infection may be associated with the development of head and neck cancers. To clarify the role of HPV in larynx carcinoma, 25 patients were studied for the presence of viral DNA, possible virus integration into the cellular genome, and viral expression both in neoplastic tissues and in neighbouring normal mucosa. Twelve of 25 patients with neoplasia (48%) showed negative results for HPV sequences, and 13 (52%) showed positive results. Among the latter group of patients, seven were HPV-16 positive, five were HPV-6, and one was HPV-45. No multiple infections were detected. The physical status of the HPV genome was analysed by three methods: polymerase chain reaction (PCR), bidimensional agarose gel electrophoresis, and in situ hybridisation. Viral integration into the host genome occurred in 43% of cases of HPV-16 and in 20% of cases of HPV-6. Viral RNA expression was detected by reverse transcription-PCR only in HPV-16-positive tumours. The pattern of expression was consistent with an active role of HPV in cellular transformation. In conclusion, the present work suggests that HPV infection may be involved in some cases of laryngeal carcinoma. However, the transformation mechanisms might be different from those currently accepted for anogenital cancers.

A glycoprotein E deletion mutant of bovine herpesvirus 1 infects the same limited number of tissues in calves as wild-type virus, but for a shorter period.

To gain insight into the role of glycoprotein E of bovine herpesvirus 1 (BHV-1), we compared the distribution of wild-type (wt) BHV-1 with that of a gE deletion mutant (gE-) in calves after intranasal inoculation. The wt-infected calves had severe clinical signs, but the gE(-)-infected calves were virtually free of clinical signs. At 3, 4, 7, 8, 44, 45, 50 and 51 days post-infection (p.i.), one calf from each group was killed and tissues were collected for virus isolation and PCR analysis. At 3, 4, 7 and 8 days p.i., infectious virus could be isolated only from the nasopharyngeal mucosa, parotid gland and nearby lymphoid tissues for both the wt- and gE(-)-infected calves. At 3 and 4 days p.i., virus titres in these tissues were comparable in both the wt- and gE(-)-infected calves. However, the virus titres were significantly reduced at 7 and 8 days p.i. in the gE(-)-infected calves, but not in the wt-infected calves. Semi-quantitative PCR analysis revealed that for the entire infection period (3 to 51 days p.i.) significantly more BHV-1 DNA was detected in the trigeminal ganglia (TG) of the wt-infected calves than in those of the gE(-)-infected calves. We conclude that the gE- mutant infects the same limited number of tissues as wt BHV-1, but for a shorter period.