Oropharynx microbiota transitions in hypopharyngeal carcinoma treatment of induced chemotherapy followed by surgery.

AIMS: To analyze changes in oropharynx microbiota composition after receiving induced chemotherapy followed by surgery for hypopharyngeal squamous cell carcinoma (HPSCC) patients. METHODS: Clinical data and swab samples of 38 HPSCC patients (HPSCC group) and 30 patients with benign disease (control group, CG) were enrolled in the study. HPSCC group was stratified into two groups: induced chemotherapy group (IC) of 10 patients and non-induced chemotherapy group (nIC) of 28 patients. The microbiota from oropharyngeal membrane was analyzed through 16S rRNA sequencing. RESULTS: Alpha-diversity (Shannon and Ace indexes) and weighted UniFrac based beta-diversity severely decreased in the HPSCC group when compared with CG. In pre-operative comparisons, PCoA and NMDS analyses showed microbial structures in the IC group were more similar to CG than nIC. Both IC group and nIC group yielded significantly diverse post-operative communities in contrast to their pre-operative counterparts, evident by the decrease in genera Veillonella and Fusobacterium and increase in genera Streptococcus and Gemella. Given that post-operative oropharynx microbiota showed no difference between IC and nIC groups, the IC group showed less accumulation in anaerobic communities. The abundance of genera Fusobacterium, Parvimonas, Actinomyces were enhanced in the advanced stages (III/IV). CONCLUSIONS: Oropharynx microbiota in the HPSCC group presents dysbiosis with low diversity and abundance. Induced chemotherapy is beneficial in adjusting the oropharynx microbial environment leading to fewer amounts of anaerobe accumulation after operation. Higher amounts of Fusobacterium in advanced stages (III/IV) may influence the progression of HPSCC.

Microbiomic differences at cancer-prone oral mucosa sites with marijuana usage.

Marijuana smoke contains cannabinoids, immunosuppressants, and a mixture of potentially-mutagenic chemicals. In addition to systemic disease, it is thought to contribute to oral disease, such as tooth loss, tissue changes in the gums and throat, and possibly oral pharyngeal cancer. We used a cross-sectional study of 20 marijuana users and 19 control non-users, to determine if chronic inhalation-based exposure to marijuana was associated with a distinct oral microbiota at the two most common sites of head and neck squamous cell carcinoma (HNSCC), the lateral border of the tongue and the oral pharynx. At the tongue site, genera earlier shown to be enriched on HNSCC mucosa, Capnocytophaga, Fusobacterium, and Porphyromonas, were at low levels in marijuana users, while Rothia, which is found at depressed levels on HNSCC mucosa, was high. At the oral pharynx site, differences in bacteria were distinct, with higher levels of Selenomonas and lower levels of Streptococcus which is what is seen in HNSCC. No evidence was seen for a contribution of marijuana product contaminating bacteria to these differences. This study revealed differences in the surface oral mucosal microbiota with frequent smoking of marijuana.

Variations in Oral Microbiota Composition Are Associated With a Risk of Throat Cancer.

In this study, a next-generation sequencing strategy on 16S ribosomal RNA (16S rRNA) gene was employed to analyze 70 oral samples from 32 patients with throat cancer, nine patients with vocal cord polyp, and 29 healthy individuals (normal controls). Using this strategy, we demonstrated, for the first time, that the salivary microbiota of cancer patients were significantly different from those of patients with a polyp and healthy individuals. We observed that the beta diversity of the cancer group was divergent from both the normal and polyp groups, while alpha-diversity indices such as the Chao1 estimator (P = 8.1e-05), Simpson (P = 0.0045), and Shannon (P = 0.0071) were significantly reduced in cancer patients compared with patients containing a polyp and normal healthy individuals. Linear discriminant analysis (LDA) and Kruskal-Wallis test analyses and real-time quantitative polymerase chain reaction (qPCR) verification test revealed that the genera Aggregatibacter, Pseudomonas, Bacteroides, and Ruminiclostridium were significantly enriched in the throat cancer group compared with the vocal cord polyp and normal control groups (score value >2). Finally, diagnostic models based on putatively important constituent bacteria were constructed with 87.5% accuracy [area under the curve (AUC) = 0.875, 95% confidence interval (CI): 0.695-1]. In summary, in this study we characterized, for the first time, the oral microbiota of throat cancer patients without smoking history. We speculate that these results will help in the pathogenic mechanism and early diagnosis of throat cancer.

Prognostic significance and association of Helicobacter pylori infection in pharyngolaryngeal cancer.

The aim of this study is to assess the correlation between the Helicobacter pylori (H. pylori) serologic status of patients who underwent for curative resection for squamous cell carcinoma of the larynx and hypopharynx and their prognosis. From April 2004 to March 2005, we included eighty patients with laryngeal and hypopharyngeal cancer. Control group consisted of 20 healthy patients and 10 patients with Reinke's edema. Serologic status was assessed using an enzyme-linked immunosorbent assay kit for immunoglobulin G. Patients were followed for 5 years. H. pylori-positive serologic status was statistically significant for the case subjects (70.6 v/s 29.4 %; p < 0.001). Mean overall and disease-free survival were 50.7 months (range 46.9-54.5) and 52.1 months (range 48.3-55.7), respectively. H. pylori-positive serologic status was not associated with a poor prognosis in the Cox regression model (p = 0.77). We observed a positive association between H. pylori infection and laryngeal and hypopharyngeal cancer. But we fail to confirm that the presence of H. pylori infection is associated with poor outcome or a higher recurrence rate.

Pilus gene pool variation and the virulence of Corynebacterium diphtheriae clinical isolates during infection of a nematode.

Toxigenic Corynebacterium diphtheriae strains cause diphtheria in humans. The toxigenic C. diphtheriae isolate NCTC13129 produces three distinct heterotrimeric pili that contain SpaA, SpaD, and SpaH, making up the shaft structure. The SpaA pili are known to mediate bacterial adherence to pharyngeal epithelial cells. However, to date little is known about the expression of different pili in various clinical isolates and their importance in bacterial pathogenesis. Here, we characterized a large collection of C. diphtheriae clinical isolates for their pilin gene pool by PCR and for the expression of the respective pilins by immunoblotting with antibodies against Spa pilins. Consistent with the role of a virulence factor, the SpaA-type pili were found to be prevalent among the isolates, and most significantly, corynebacterial adherence to pharyngeal epithelial cells was strictly correlated with isolates that were positive for the SpaA pili. By comparison, the isolates were heterogeneous for the presence of SpaD- and SpaH-type pili. Importantly, using Caenorhabditis elegans as a model host for infection, we show here that strain NCTC13129 rapidly killed the nematodes, the phenotype similar to isolates that were positive for toxin and all pilus types. In contrast, isogenic mutants of NCTC13129 lacking SpaA-type pili or devoid of toxin and SpaA pili exhibited delayed killing of nematodes with similar kinetics. Consistently, nontoxigenic or toxigenic isolates that lack one, two, or all three pilus types were also attenuated in virulence. This work signifies the important role of pili in corynebacterial pathogenesis and provides a simple host model to identify additional virulence factors.

Attachment and invasion of Neisseria meningitidis to host cells is related to surface hydrophobicity, bacterial cell size and capsule.

We compared exemplar strains from two hypervirulent clonal complexes, strain NMB-CDC from ST-8/11 cc and strain MC58 from ST-32/269 cc, in host cell attachment and invasion. Strain NMB-CDC attached to and invaded host cells at a significantly greater frequency than strain MC58. Type IV pili retained the primary role for initial attachment to host cells for both isolates regardless of pilin class and glycosylation pattern. In strain MC58, the serogroup B capsule was the major inhibitory determinant affecting both bacterial attachment to and invasion of host cells. Removal of terminal sialylation of lipooligosaccharide (LOS) in the presence of capsule did not influence rates of attachment or invasion for strain MC58. However, removal of either serogroup B capsule or LOS sialylation in strain NMB-CDC increased bacterial attachment to host cells to the same extent. Although the level of inhibition of attachment by capsule was different between these strains, the regulation of the capsule synthesis locus by the two-component response regulator MisR, and the level of surface capsule determined by flow cytometry were not significantly different. However, the diplococci of strain NMB-CDC were shown to have a 1.89-fold greater surface area than strain MC58 by flow cytometry. It was proposed that the increase in surface area without changing the amount of anchored glycolipid capsule in the outer membrane would result in a sparser capsule and increase surface hydrophobicity. Strain NMB-CDC was shown to be more hydrophobic than strain MC58 using hydrophobicity interaction chromatography and microbial adhesion-to-solvents assays. In conclusion, improved levels of adherence of strain NMB-CDC to cell lines was associated with increased bacterial cell surface and surface hydrophobicity. This study shows that there is diversity in bacterial cell surface area and surface hydrophobicity within N. meningitidis which influence steps in meningococcal pathogenesis.

[A comparative analysis of antibiotics efficacy in acute ENT infections].

The comparison of the efficacy of a number of antibacterial drugs in acute pyoinflammatory ENT diseases allowed the authors to recommend moxifloxacin as most effective in acute purulent ENT pathology. Moxifloxacin demonstrated its superiority in seeding (the highest sensitivity of the bacteria to this antibiotic), in the speed of development of the bactericidal action and in treatment results. The drug had no significant side effects.

Effect of preoperative prophylaxis with filgrastim in cancer neck dissection.

BACKGROUND: Cancer surgery is known to lead to a deterioration in host defence mechanisms and an increase in susceptibility to infection after operation. Filgrastim enhances important antimicrobial functions of neutrophils including chemotaxis, phagocytosis and oxidative killing mechanisms. METHODS: The effects of additional (all patients received perioperative 3 ' 25 mg kg-1 cefotiam and 1 ' 20 mg kg-1 metronidazole) preoperative prophylaxis with filgrastim (5 microg kg-1 12 h prior to surgery plus 5 microg kg-1 0 h prior to surgery) on neutrophil phagocytosis and reactive oxygen radical production and postoperative infections in 24 patients undergoing cancer neck dissection were studied. Phagocytic capacity was assessed by measuring the uptake of fluorescein isothiocyanate-labelled Escherichia coli and Staphylococcus aureus by flow cytometry. Reactive oxygen generation after phagocytosis was estimated by determining the amount of dihydrorhodamine 123 converted to rhodamine 123, intracellularly. RESULTS: In the filgrastim-treated patients a higher neutrophil phagocytic capacity was seen intraoperatively, and 1-5 days postoperative, but not prior to surgery. Reactive oxygen radical production was significantly higher in filgrastim-treated patients prior to surgery, intraoperative and postoperative (1-5 days). 2/12 (17%) patients had postoperative infections in the filgrastim group and 9/12 (75%) patients had infections in the placebo group (P < 0.001). In particular, wound infections were recorded more often in the placebo group (1/12 vs. 6/12; P = 0.004). CONCLUSION: We conclude that filgrastim enhances perioperative neutrophil function and could be useful in the prophylaxis of postoperative wound infections in patients undergoing cancer neck dissection.

Determination of nonendemic nasopharyngeal carcinoma by in situ hybridization for Epstein-Barr virus EBER1 RNA: sensitivity and specificity in cervical node metastases.

After time-consuming and costly investigations, patients with neck metastases from an occult primary often receive unnecessarily large radiation volumes to treat a possible origin in the nasopharynx. In this study a colorimetric antisense Epstein-Barr early ribonucleoprotein 1 (EBER1) oligonucleotide probe specific for Epstein-Barr virus RNA was hybridized in situ to metastatic tissue obtained from 18 nasopharyngeal, 54 oral and pharyngeal, and 12 occult carcinomas derived from an unselected population. All 16 nonkeratinizing nasopharyngeal carcinomas (NPCs) were positive for EBER1. Both cases of keratinizing NPC and all 54 other metastases were negative. A single positive case of occult carcinoma indicated its origin from NPC. In retrospect, 7 patients with occult carcinoma had received unnecessary treatment with irradiation to the nasopharynx. Nasopharyngeal carcinoma appears to be a less common origin of occult carcinoma than previously considered. In the proper clinicopathologic context EBER1 in situ hybridization (EBER1-ISH) allows exclusion of NPC with a high degree of accuracy. Thus unnecessarily large radiation volumes and their adverse sequelae may be reduced in the treatment of occult carcinoma. Conversely, a positive result of ISH allows exclusion of further extensive diagnostic procedures.

Comparison of Virapap filter hybridization with polymerase chain reaction and Southern blot hybridization methods for detection of human papillomavirus in tonsillar and pharyngeal cancers.

A method using a commercial dot filter hybridization kit, Virapap, was compared with Southern blot hybridization and polymerase chain reaction (PCR) for the detection of human papillomavirus (HPV) types 16 and 18 in pharyngeal and tonsillar cancers of 12 patients as well as tonsillar biopsies from 28 patients with chronic tonsillitis. Concordant results between Virapap and PCR, Virapap and Southern hybridization, and PCR and Southern hybridization methods were obtained respectively in 41.7%, 58.3% and 83.3% of the cancer cases, and 67.9%, 67.9% and 85.7% of the control (tonsillitis) cases. Virapap false-positive results were found in 5 cancer cases and 5 control cases. Although the Virapap method is reported to be useful for detecting HPV in gynecological tissues, this method cannot be recommended for the detection of HPV in pharyngeal and tonsillar cancers.

Presence of human papillomavirus type 18 DNA in a pharyngeal and a laryngeal carcinoma.

We examined the presence of human papillomavirus (HPV) genome in pharyngeal and laryngeal squamous cell carcinomas by employing the polymerase chain reaction. We detected HPV 16 DNA in one of 11 pharyngeal carcinomas and in 3 of 28 laryngeal carcinomas, and HPV 18 DNA in a pharyngeal and a laryngeal carcinoma. In one of the laryngeal carcinomas, DNA of both HPV types 16 and 18 were detected. To our knowledge, this is the first report of detection of HPV type 18 in head and neck carcinomas.

Detection of human papillomavirus in head and neck tumors with DNA hybridization and immunohistochemical analysis.

The presence of human papillomavirus (HPV) DNA in oral, sinus, pharynx, and larynx lesions of Japanese patients was studied by Southern blot hybridization under less stringent (25% formamide, 42 degrees C) and stringent (50% formamide, 42 degrees C) conditions. Three samples from 10 benign tumors, and 3 of 30 malignant tumors, contained HPV DNA or HPV-related sequences. The HPV DNAs harbored in three laryngeal papillomas were HPV-11, -6, and -6 or -11, respectively. The HPV DNA and viral capsid antigens were easily detected by in situ hybridization, Western blotting, and peroxidase-antiperoxidase staining. However, neither the typical restriction pattern of HPV DNA nor viral antigen was identified in the malignant tumors, suggesting that subgenomic fragments remained integrated in the host cell DNA.

[Bacterial growth on carcinomas of the oral mucosa]. / Zur bakteriellen Besiedlung von Karzinomen der Mundschleimhaut.

Carcinomas of the oral mucosa are colonized by a variety of aerobic and anaerobic bacteria. Apart from the normal flora of the oral cavity also organisms of pathogenic potential are found, which may cause postoperative wound infections. A specific antibiotic prophylaxis regimen in extensive head and neck surgery requires identification of the pathopotent organisms and sensitivity testing followed by perioperative individual antimicrobial chemoprophylaxis management. The results of an additional bacteriological examination of the surrounding pharyngeal flora showed no major difference to the range of organisms found in the carcinoma.

Association of Epstein-Barr virus with polymorphic reticulosis.

We report a case of polymorphic reticulosis (PR) associated with pharyngeal replication of Epstein-Barr virus (EBV). A 78-year-old man with necrotic inflammatory granulations in the nasal cavities and ulcerative tissue of the tonsils was found to have PR after an initial diagnosis of lethal midline granuloma and was found to have high antibody titers to EBV. EBV-determined nuclear antigen (EBNA) was demonstrated in pharyngeal biopsy specimens by the anti-complement immunofluorescent technique, following which dual immunofluorescence staining, EBNA and T-cell antigen analysis were performed, using a wide variety of monoclonal antibodies. All of the EBNA-positive lymphocytes in the pharyngeal biopsy specimens were found to have exclusively T-cell antigens. This case strongly suggests that some of the cases of PR related to T-cell lymphomas may be closely associated with active EBV infection.

Epstein-Barr virus receptors on human pharyngeal epithelia.

The apparently strict tropism of Epstein-Barr virus (EBV) for B lymphocytes has been attributed to the existence of a B-lineage-specific surface molecule, the C3d receptor, which also functions as a receptor for EBV. Two monoclonal antibodies against different determinants on the EBV/C3d receptor of B cells were shown to react with pharyngeal epithelia in a cell differentiation-dependent manner. These findings, which raise the possibility of direct virus entry into a naturally exposed epithelium, strengthen the evidence in favour of an epithelial reservoir of EBV infection in vivo and identify a means whereby the virus/epithelium interactions leading to nasopharyngeal carcinoma might be initiated.

Microfilariae in laryngeal and pharyngeal brushing smears from a case of carcinoma of the pharynx.

Microfilariae were observed in May-Grünwald-Giemsa-stained brushing smears of the laryngeal and pharyngeal areas in a 35-year-old man with carcinoma of the pharyngeal wall and marked laryngeal edema. Liberation of microfilariae due to lymphorrhexis following severe lymphedema was considered to be the mechanism for the infestation of these sites, which has not been previously reported.

[Microflora of the suppurative inflammatory foci in patients with laryngeal and pharyngeal cancer undergoing combined treatment for local postoperative complications]. / Mikroflora gnoino-vospalitel'nykh ochagov u bol'nykh rakom gortani i glotki pri kompleksnom lechenii mestnykh posleoperatsionnykh oslozhnenii.

The paper deals with the problems of treatment for local postoperative complications in patients suffering cancer of the larynx and throat. Early diagnosis of complications as well as therapy employing modern glass fibre equipment are discussed. The first attempt to study the microflora of the newly formed throat in the early postoperative period is described. A wide spectrum of dormant bacteria was identified. Healing was found to depend upon microorganism level and profile of bacterial species in the newly formed throat. Complex application of endoscopic and laser procedures proved to be an effective means of sanative care.