Role of Actinomyces spp. and related organisms in the development of medication-related osteonecrosis of the jaw (MRONJ): Clinical evidence based on a case series.

Medication-related osteonecrosis of the jaw (MRONJ) is an increasingly common consequence of antiresorptive treatment, which often leads to the development of necrotic exposed bone surfaces with inflammatory processes affecting the jawbone. Although the development of MRONJ is often associated with the inflammatory response or infections caused by the colonizing members of the oral microbiota, the exact pathogenesis of MRONJ is still not fully understood. In the present paper, we aimed to provide additional, microbiological culture-supported evidence, supporting the "infection hypothesis" that Actinomyces spp. and related organisms may play an important pathogenic role in the development of MRONJ and the resulting bone necrosis. In our case series, all patients presented with similar underlying conditions and anamnestic data, and have received antiresorptive medications (bisphosphonates or a RANK ligand (RANKL) inhibitor) to prevent the occurrence or progression of bone metastases, secondary to prostate cancer. Nevertheless, a few years into antiresorptive drug therapy, varying stages of MRONJ was identified in the mentioned patients. In all three cases, quantitative microbiological culture of the necrotic bone samples yielded a complex microbiota, dominated by Actinomyces and Schaalia spp. with high colony counts. Additionally, our followed-up case series document the treatment of these patients with a combination of surgical intervention and long-term antibiotic therapy, where favourable clinical responses were seen is all cases. If the "infection hypothesis" is valid, it may have significant consequences in the preventative and therapeutic strategies associated with this disease.

The role of Actinomyces spp. and related organisms in cervicofacial infections: Pathomechanism, diagnosis and therapeutic aspects.

Members of the Actinomyces genus and Actinomyces-like organisms (ALOs; namely Actinotignum, Arcanobacterium, Schaalia and Varibaculum) are Gram-positive, non-spore-forming rods that are commensal members of the human oral cavity, gastrointestinal tract, female genital tract and skin microbiota. Cervicofacial actinomycosis or "lumpy jaw syndrome" - the chronic, suppurative granulomatous disease caused by Actinomyces spp. And ALOs - is characterized by an initially slow and unspecific disease-presentation, which often mimics other pathologies, followed by the formation of painful abscesses and severe tissue destruction. Actinomycosis has been described as a rare disease, however, reliable epidemiological data are lacking. In addition, there is increasing awareness regarding the role of Actinomyces spp. in the development of osteoradionecrosis and medication-related osteonecrosis of the jaw. The aim of this narrative review is to succinctly summarize the current advances regarding the microbiological, clinical, diagnostic and therapeutic aspects of cervicofacial actinomycosis, in addition to the roles of Actinomyces species and ALOs as members of the oral microbiota and in dental biofilm, in other dental infections (caries, root canal infection, periapical infection, periodontitis) and osteonecrosis of the jaw, in the context of recent taxonomic changes affecting the genus. Our paper aims to be a blueprint for dentists, other physicians, microbiologists and researchers regarding the multifaceted field of cervicofacial actinomycosis.

Incidence and Clinical Characteristics of Anaerobic Bacteremia at a University Hospital in Hungary: A 5-Year Retrospective Observational Study.

Strict anaerobes have been reported to account for 0.5-13% of episodes of bacteremia in the adult population, with a growing awareness among clinicians regarding anaerobic bacteremia, especially in patients with specific predisposing factors. The aim of our present study was to assess the incidence and clinical characteristics of anaerobic bacteremia during a 5-year period (2016-2020) at a tertiary care teaching hospital, and to compare our findings with other studies in Hungary. Overall, n = 160 strict anaerobes were detected, out of which, 44.4% (n = 71; 0.1% of positive blood cultures, 0.1/1000 hospitalizations, 3.3/100,000 patient days) were clinically significant, while Cutibacterium spp. accounted for 55.6% (n = 89) of isolates. Among relevant pathogens, the Bacteroides/Parabacteroides spp. group (32.4%; n = 23), Clostridium spp. (22.5%; n = 16) and Gram-positive anaerobic cocci (15.5%; n = 11) were the most common. The mean age of patients was 67.1 ± 14.1 years, with a male majority (59.2%; n = 42). A total of 38.0% of patients were affected by a malignancy or immunosuppression, while an abscess was identified in 15.5% of cases. A total of 74.7% (n = 53) of patients received antibiotics prior to blood culture sampling; in instances where antimicrobials were reported, anaerobic coverage of the drugs was appropriate in 52.1% (n = 37) of cases. The 30-day crude mortality rate was 39.4% (n = 28); age ≥ 75 years was a significant predictor of 30-day mortality (OR: 5.0; CI: 1.8-14.4; p = 0.003), while malignancy and immunosuppression, lack of anti-anaerobic coverage or female sex did not show a significant relationship with the mortality of these patients. Early recognition of the role played by anaerobes in sepsis and timely initiation of adequate, effective antimicrobial treatment have proven efficient in reducing the mortality of patients affected by anaerobic bacteremia.

Tumour Necrosis Factor Alpha-induced Protein 3 Negatively Regulates Cutibacterium acnes-induced Innate Immune Events in Epidermal Keratinocytes.

Human epidermal keratinocytes sense the presence of human skin microbiota through pathogen recognition receptors, such as toll-like receptors, and induce innate immune and inflammatory events. In healthy epidermis there is an absence of inflammation despite the continuous presence of cutaneous microbes, which is evidence of an effective immune regulatory mechanism. The aim of this study was to investigate tumour necrosis factor alpha-induced protein 3 (TNFAIP3), a negative regulator of toll-like receptor and nuclear factor kappa B signalling pathways, and its role in these regulatory events. A broad spectrum of toll-like receptor ligands induced TNFAIP3 expression, as did live Cutibacterium acnes, which is involved in the pathogenesis of acne. Changes in bacterium-induced, dose-dependent TNFAIP3 expression were Jun kinase- and nuclear factor kappa B-dependent, and resulted in altered cytokine and chemokine levels in in vitro cultured human keratinocytes. In acne lesions, TNFAIP3 mRNA expression was elevated compared with non-lesional skin samples from the same individuals. These results suggest that TNFAIP3 may have a general role in fine regulation of microbiota-induced cutaneous immune homeostasis.

Effects of different decontaminating solutions used for the treatment of peri-implantitis on the growth of Porphyromonas gingivalis-an in vitro study.

Implants have been considered the treatment of choice to replace missing teeth, unfortunately, peri-implant disease is still an unresolved issue. Contaminated implants may be decontaminated by physical debridement and chemical disinfectants; however, there is a lack of consensus regarding the ideal techniques/agents to be used for the decontamination. The objective of our study was to compare the decontaminating efficacy of different chemical agents on a titanium surface contaminated with Porphyromonas gingivalis, a typical representative of the bacterial flora associated with peri-implantitis. Commercially pure Ti grade 4 discs with a polished surface were treated with a mouthwash containing chlorhexidine digluconate (0.1%), povidone-iodine (PVP-iodine) solution (10%) or citric acid monohydrate (40%). Qualitative and quantitative assessment of cellular growth and survival were assessed by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and scanning electron microscopy (SEM). Significant differences in the quantity of P. gingivalis could be observed after 6 days of incubation. A numerical, but not statistically significant (P = 0.066) decrease in the amount of living bacteria was observed in the group treated with the PVP-iodine solution as compared to the control group. The chlorhexidine (CHX)-treated group presented with significantly higher cell counts, as compared to the PVP-iodine-treated group (P = 0.032), while this was not observed compared to the control group and citric acid-treated group. Our results have also been verified by SEM measurements. Our results suggest that for P. gingivalis contamination on a titanium surface in vitro, PVP-iodine is a superior decontaminant, compared to citric acid and chlorhexidine-digulconate solution.

The Pathogenic Role of Actinomyces spp. and Related Organisms in Genitourinary Infections: Discoveries in the New, Modern Diagnostic Era.

Actinomycosis is a chronic, suppurative, granulomatous infectious disease, caused by different species of Actinomyces bacteria. To date, 26 validly published Actinomyces species have been described as part of a normal human microbiota or from human clinical specimens. Due to the rapid spread of new, modern diagnostic procedures, 13 of 26 of these species have been described in this century and the Actinomycetaceae family has undergone several taxonomic revisions, including the introduction of many novel species termed Actinomyces-like organisms (ALOs). There is scarce data available on the role of these novel bacterial species in various infectious processes in human medicine. The aim of this review is to provide a comprehensive overview of Actinomyces and closely related organisms involved in human diseases-with a special focus on newly described species-in particular their role in genitourinary tract infections in females and males.

Cutibacterium acnes regulates the epidermal barrier properties of HPV-KER human immortalized keratinocyte cultures.

Our skin provides a physical barrier to separate the internal part of our body from the environment. Maintenance of complex barrier functions is achieved through anatomical structures in the skin, the stratified squamous epithelium specialized junctional organelles, called tight junctions (TJs). Several members of our microbial communities are known to affect the differentiation state and function of the colonized organ. Whether and how interactions between skin cells and cutaneous microbes, including Cutibacterium acnes (C. acnes), modify the structure and/or function of our skin is currently only partly understood. Thus, in our studies, we investigated whether C. acnes may affect the epidermal barrier using in vitro model systems. Real-time cellular analysis showed that depending on the keratinocyte differentiation state, the applied C. acnes strains and their dose, the measured impedance values change, together with the expression of selected TJ proteins. These may reflect barrier alterations, which can be partially restored upon antibiotic-antimycotic treatment. Our findings suggest that C. acnes can actively modify the barrier properties of cultured keratinocytes, possibly through alteration of tight cell-to-cell contacts. Similar events may play important roles in our skin, in the maintenance of cutaneous homeostasis.

Streptococcus suis: An Underestimated Emerging Pathogen in Hungary?

Streptococcus suis (S. suis) is an emerging zoonotic pathogen, demonstrated as an etiological agent in human infections in increasing frequency, including diseases like purulent meningitis, sepsis, uveitis-endophtalmitis and arthritis. Due to the increased availability and utility of novel diagnostic technologies in clinical microbiology, more studies have been published on the epidemiology of S. suis, both in veterinary and human medicine; however, there are no comprehensive data available regarding human S. suis infections from East-Central European countries. As a part of our study, data were collected from the National Bacteriological Surveillance (NBS) system on patients who had at least one positive microbiological result for S. suis, corresponding to an 18-year study period (2002-2019). n = 74 S. suis strains were isolated from invasive human infections, corresponding to 34 patients. The number of affected patients was 1.89 ± 1.53/year (range: 0-5). Most isolates originated from blood culture (63.5%) and cerebrospinal fluid (18.9%) samples. Additionally, we present detailed documentation of three instructive cases from three regions of the country and with three distinctly different outcomes. Hungary has traditional agriculture, the significant portion of which includes the production and consumption of pork meat, with characteristic preparation and consumption customs and unfavorable epidemiological characteristics (alcohol consumption, prevalence of malignant diseases or diabetes), which have all been described as important predisposing factors for the development of serious infections. Clinicians and microbiologist need to be vigilant even in nonendemic areas, especially if the patients have a history of occupational hazards or having close contact with infected pigs.

Characterization of a rare bla VIM-4 metallo-ß-lactamase-producing Serratia marcescens clinical isolate in Hungary.

A carbapenem-resistant S. marcescens isolate was recovered from a patient with an inflammed pacemaker inplantation pocket from a Cardiac Surgery ward in a Hungarian University Hospital. Phenotypic tests and polymerase chain reaction (PCR) confirmed a very rare gene responsible for production of a carbapenemase ( bla VIM-4 ), which was further characterized by Sanger-sequencing. The characterization of this S. marcescens strain emphasizes the ongoing emergence of novel or rare carbapenemases. Strains expressing a weak carbapenemase like this strain might go unrecognized by routine diagnostics due to low minimum inhibitory concentrations (MICs) for the bacterial strains producing such enzymes.

A 10-year single-center experience on Stenotrophomonas maltophilia resistotyping in Szeged, Hungary.

Stenotrophomonas maltophilia is an aerobic, oxidase-negative and catalase-positive bacillus. S. maltophilia is a recognized opportunistic pathogen. Due to the advancements in invasive medical procedures, organ transplantation and chemotherapy of malignant illnesses, the relevance of this pathogen increased significantly. The therapy of S. maltophilia infections is challenging, as these bacteria show intrinsic resistance to multiple classes of antibiotics, the first-choice drug is sulfamethoxazole/trimethoprim. Our aim was to assess the epidemiology of S. maltophilia from various clinical samples and the characterization of resistance-levels and resistotyping of these samples over a long surveillance period. The study included S. maltophilia bacterial isolates from blood culture samples, respiratory samples and urine samples and the data for the samples, received between January 2008 until December 2017, a total of 817 S. maltophilia isolates were identified (respiratory samples n = 579, 70.9%, blood culture samples n = 175, 21.4% and urine samples n = 63, 7.7%). Levofloxacin and colistin-susceptibility rates were the highest (92.2%; n = 753), followed by tigecycline (90.5%, n = 739), the first-line agent sulfamethoxazole/trimethoprim (87.4%, n = 714), while phenotypic resistance rate was highest for amikacin (72.5% of isolates were resistant, n = 592). The clinical problem of sulfamethoxazole/trimethoprim-resistance is a complex issue, because there is no guideline available for the therapy of these infections.

Fatal case of bacteremia caused by Streptococcus suis in a splenectomized man and a review of the European literature.

Streptococcus suis is an emerging zoonotic human pathogen, which is a causative agent of invasive infections in people who are in close contact with infected pigs or contaminated pork products. It is associated with severe systemic infections, most commonly meningitis and sepsis, which may lead to high rates of morbidity and mortality. Serotype 2 is the most prevalent type in S. suis infections in humans. We have reported a case of a very rapidly proceeding fatal human S. suis infection in a splenectomized, but otherwise immunocompetent patient in Hungary. We would like to highlight the attention for this pathogen for the risk group patients, not only pig breeders, veterinarians, abattoir workers, meat processing and transport workers, butchers and cooks, that those persons who are immunocompromised including those with spleen removed, persons with diabetes mellitus, cancer and alcoholism, are also at greater risk of infection.

Microbiological and Clinical Aspects of Cervicofacial Actinomyces Infections: An Overview.

Similarly to other non-spore-forming Gram-positive anaerobes, members of the Actinomyces genus are important saprophytic constituents of the normal microbiota of humans. Actinomyces infections are considered to be rare, with cervicofacial infections (also known as 'lumpy jaw syndrome') being the most prevalent type in the clinical practice. Actinomycoses are characterized by a slowly progressing (indolent) infection, with non-specific symptoms, and additionally, the clinical presentation of the signs/symptoms can mimic other pathologies, such as solid tumors, active Mycobacterium tuberculosis infections, nocardiosis, fungal infections, infarctions, and so on. The clinical diagnosis of actinomycosis may be difficult due to its non-specific symptoms and the fastidious, slow-growing nature of the pathogens, requiring an anaerobic atmosphere for primary isolation. Based on 111 references, the aim of this review is to summarize current advances regarding the clinical features, diagnostics, and therapy of cervicofacial Actinomyces infections and act as a paper for dentistry specialists, other physicians, and clinical microbiologists.

Prevalence and Antibiotic Resistance of Stenotrophomonas maltophilia in Respiratory Tract Samples: A 10-Year Epidemiological Snapshot.

BACKGROUND: Since the 1980s, Stenotrophomonas maltophilia has emerged as an important pathogen associated with significant mortality in pneumonia and bacteremia of severely immunocompromised, hospitalized patients. The drug of choice in S maltophilia infections is sulfamethoxazole-trimethoprim (SMX/TMP); SMX/TMP resistance is a serious concern in clinical practice. The aim of this study was to assess the prevalence of S maltophilia in lower respiratory tract (LRTI) samples at a tertiary-care university hospital. METHODS: This retrospective cohort study was carried out using microbiological data collected between January 2008 and December 2017. Routine antimicrobial susceptibility testing was performed for SMX/TMP and levofloxacin; in case of resistance, susceptibility testing for additional antibiotics (tigecycline, amikacin, and colistin) was also performed. RESULTS: A total of 579 individual S maltophilia isolates were identified (2008-2012: n = 160, 2013-2017: n = 419; P = .0008). In all, 78.46% of patients were younger than 5 or older than 50 years of age and had recent trauma, surgery, or underlying conditions (malignancies, respiratory distress syndrome, congenital disorders, and cystic fibrosis). In 28.16% of samples, more than 1 pathogen was identified, and 5.35% of coisolated pathogens were multidrug resistant (MDR). In all, 12.1% of isolates were SMX/TMP-resistant (2008-2012: 6.12%, 2013-2017: 18.06%; P = .034), while 8.99% were resistant to levofloxacin (2008-2012: 7.86%, 2013-2017: 10.12%; P > .05). SMX/TMP resistance was detected more frequently in samples originating from inpatients (n = 2.50 ± 2.39 vs n = 11.50 ± 3.76; P = .0002). CONCLUSIONS: In all, 5.87% of isolates were extensively drug resistant (XDR), that is, in addition to SMX/TMP, they were resistant to levofloxacin, amikacin, colistin, and tigecycline. The results of our study correspond to the findings in the literature.

Epidemiological Trends and Resistance Associated with Stenotrophomonas maltophilia Bacteremia: A 10-Year Retrospective Cohort Study in a Tertiary-Care Hospital in Hungary.

Stenotrophomonas maltophilia has been recognized as an emerging nosocomial pathogen in invasive infections of immunocompromised, severely debilitated patients with significant underlying illnesses. The first-choice drug in these infections is sulfamethoxazole-trimethoprim (SMX/TMP), and resistance to this antimicrobial is a daunting challenge for clinicians. The aim of this study was to assess the prevalence of S. maltophilia bacteremia and SMX/TMP-resistance levels at a tertiary-care university hospital. A total of 175 episodes of S. maltophilia bacteremia were identified (2008-2012: n = 82, 2013-2017: n = 93; p = 0.061), 52% of affected patients were 60 years of age, and had recent surgery, severe injuries or underlying conditions (malignant hematologic diseases and solid tumors) in their history. Sixteen percent of isolates were resistant to SMX/TMP (2008-2012: n = 13.8%, 2013-2017: n = 17.2%; p = 0.076), and out of the resistant strains, 32.7% were also resistant to levofloxacin and colistin. Our findings on the SMX/TMP-resistance were similar to global literature data.

Chicken or the Egg: Microbial Alterations in Biopsy Samples of Patients with Oral Potentially Malignant Disorders.

Oral carcinogenesis often leads to the alteration of the microbiota at the site of the tumor, but data are scarce regarding the microbial communities of oral potentially malignant disorders (OPMDs). Punch biopsies were taken from healthy and non-healthy mucosa of OPMD patients to analyze the microbiome using metagenome sequencing. In healthy oral mucosa biopsies the bacterial phyla Firmicutes, Fusobacteria, Proteobacteria, Actinobacteria and Bacteroidetes were detected by Ion Torrent sequencing. The same phyla as well as the phyla Fibrobacteres and Spirochaetes were present in the OPMD biopsies. On the species level, there were 10 bacterial species unique to the healthy tissue and 35 species unique to the OPMD lesions whereas eight species were detected in both samples. We observed that the relative abundance of Streptococcus mitis decreased in the OPMD lesions compared to the uninvolved tissue. In contrast, the relative abundance of Fusobacterium nucleatum, implicated in carcinogenesis, was elevated in OPMD. We detected markedly increased bacterial diversity in the OPMD lesions compared to the healthy oral mucosa. The ratio of S. mitis and F. nucleatum are characteristically altered in the OPMD lesions compared to the healthy mucosa.

TNIP1 Regulates Cutibacterium acnes-Induced Innate Immune Functions in Epidermal Keratinocytes.

Human skin cells recognize the presence of the skin microbiome through pathogen recognition receptors. Epidermal keratinocytes are known to activate toll-like receptors (TLRs) 2 and 4 in response to the commensal Cutibacterium acnes (C. acnes, formerly known as Propionibacterium acnes) bacterium and subsequently to induce innate immune and inflammatory events. These events may lead to the appearance of macroscopic inflammatory acne lesions in puberty: comedos, papules and, pustules. Healthy skin does not exhibit inflammation or skin lesions, even in the continuous presence of the same microbes. As the molecular mechanism for this duality is still unclear, we aimed to identify factors and mechanisms that control the innate immune response to C. acnes in keratinocytes using a human immortalized keratinocyte cell line, HPV-KER, normal human keratinocytes (NHEK) and an organotypic skin model (OSM). TNIP1, a negative regulator of the NF-κB signaling pathway, was found to be expressed in HPV-KER cells, and its expression was rapidly induced in response to C. acnes treatment, which was confirmed in NHEK cells and OSMs. Expression changes were not dependent on the C. acnes strain. However, we found that the extent of expression was dependent on C. acnes dose. Bacterial-induced changes in TNIP1 expression were regulated by signaling pathways involving NF-κB, p38, MAPKK and JNK. Experimental modification of TNIP1 levels affected constitutive and C. acnes-induced NF-κB promoter activities and subsequent inflammatory cytokine and chemokine mRNA and protein levels. These results suggest an important role for this negative regulator in the control of bacterially induced TLR signaling pathways in keratinocytes. We showed that all-trans retinoic acid (ATRA) induced elevated TNIP1 expression in HPV-KER cells and also in OSMs, where TNIP1 levels increased throughout the epidermis. ATRA also reduced constitutive and bacterium-induced levels of TNFα, CCL5 and TLR2, while simultaneously increasing CXCL8 and TLR4 expression. Based on these findings, we propose that ATRA may exhibit dual effects in acne therapy by both affecting the expression of the negative regulator TNIP1 and attenuating TLR2-induced inflammation. Overall, TNIP1, as a possible regulator of C. acnes-induced innate immune and inflammatory events in keratinocytes, may play important roles in the maintenance of epidermal homeostasis.

Antiproliferative and Antimicrobial Activities of Selected Bryophytes.

One-hundred and sixty-eight aqueous and organic extracts of 42 selected bryophyte species were screened in vitro for antiproliferative activity on a panel of human gynecological cancer cell lines containing HeLa (cervix epithelial adenocarcinoma), A2780 (ovarian carcinoma), and T47D (invasive ductal breast carcinoma) cells using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and for antibacterial activity on 11 strains using the disc-diffusion method. A total of 99 extracts derived from 41 species exerted ≥25% inhibition of proliferation of at least one of the cancer cell lines at 10 µg/mL. In the cases of Brachythecium rutabulum, Encalypta streptocarpa, Climacium dendroides, Neckera besseri, Pleurozium schreberi, and Pseudoleskeella nervosa, more than one extract was active in the antiproliferative assay, whereas the highest activity was observed in the case of Paraleucobryum longifolium. From the tested families, Brachytheciaceae and Amblystegiaceae provided the highest number of antiproliferative extracts. Only 19 samples of 15 taxa showed moderate antibacterial activity, including the most active Plagiomnium cuspidatum, being active on 8 tested strains. Methicillin-resistant Staphylococcus aureus (MRSA) and Staphylococcus aureus were the most susceptible to the assayed species. This is the first report on the bioactivities of these 14 species.

The first characterized carbapenem-resistant Bacteroides fragilis strain from Croatia and the case study for it.

An imipenem-resistant Bacteroides fragilis strain was isolated from the blood of a 72-year-old male patient with a urinary bladder tumor in Osijek, Croatia. This strain was also resistant to ampicillin, piperacillin/tazobactam, cefoxitin, clindamycin, tetracycline, and harbored cfiA, ermF, and tetQ genes where the high-level expression of the cfiA carbapenem-resistant gene was driven by an IS1187 element. Interestingly, despite the carbapenem-resistant feature of the B. fragilis from blood, the patient relatively easily recovered from the bacteremia. It was the first characterized imipenem-resistant B. fragilis isolate with its case report from Croatia, which confirmed the appearance of carbapenem-resistant B. fragilis strains, that continues worldwide with low incidence and the molecular characteristics vary temporally and geographically.

Prevalence and genotypes of human papillomavirus in saliva and tumor samples of head and neck cancer patients in Hungary.

In addition to traditional risk factors such as smoking, alcohol consumption and betel nut use, human papillomavirus (HPV) infection also plays a role in the development of head and neck squamous cell carcinomas (HNSCCs). Although among European countries the highest incidence and mortality rates of head and neck cancer types were recorded in Hungary, data regarding HPV prevalence in HNSCCs is scarce. We collected biopsy and saliva samples from patients diagnosed with HNSCC or oral potentially malignant disorders (OPMDs) and tested them for the presence of HPV using the PCR consensus primer set MY09/11 and the GP5+/6+ primer pair. HPV genotypes were assessed by sequencing of the amplified PCR fragments. Oral mucosa and saliva samples from tumor- and OPMD-free individuals were also analysed. HPV was detected in 11 out of 60 HNSCC samples (18%). All of the HPV positive tumors carried HPV type 16. 5 out of the 57 saliva samples collected from HNSCC patients was HPV positive (8.8%); among them, in addition to HPV16, HPV13 was also detected. Tumors located to the oropharynx had the highest HPV positivity rate with 50% (7 out of 14), which was significantly higher than the HPV prevalence in oral mucosa samples collected from controls (0 out of 20; p > 0.001) or in OPMD biopsies (0 out of 21, p > 0.001). 2 out of 57 control saliva samples (3.5%, subtype HPV13 and 11) and 3 out of 39 saliva samples from OPMD patients (7.7%, subtype HPV18, 81 and 10) were HPV positive. Our data suggested that HPV16 infection may contribute, in concert with cigarette smoking, to the development of a subset of head and neck cancers in Hungary. HPV16 infection per se does not account, however, for the high HNSCC incidence rate recorded in this country.

Propionibacterium acnes Induces Autophagy in Keratinocytes: Involvement of Multiple Mechanisms.

Propionibacterium acnes is a dominant member of the cutaneous microbiota. Herein, we evaluate the effects of different P. acnes strains and propionic acid on autophagy in keratinocytes. Our results showed that P. acnes strain 889 altered the architecture of the mitochondrial network; elevated the levels of microtubule-associated protein 1 light chain 3B-II, Beclin-1, and phospho-5'-adenosine-monophosphate-activated protein kinase α; stimulated autophagic flux; facilitated intracellular redistribution of microtubule-associated protein 1 light chain 3B; increased average number of autophagosomes per cell; and enhanced development of acidic vesicular organelles in the HPV-KER cell line. Propionic acid increased the level of phospho-5'-adenosine-monophosphate-activated protein kinase α, enhanced lipidation of microtubule-associated protein 1 light chain 3B, stimulated autophagic flux, and facilitated translocation of microtubule-associated protein 1 light chain 3B into autophagosomes in HPV-KER cells. P. acnes strains 889 and 6609 and heat-killed strain 889 also stimulated autophagosome formation in primary keratinocytes to varying degrees. These results indicate that cell wall components and secreted propionic acid metabolite of P. acnes evoke mitochondrial damage successively, thereby triggering 5'-adenosine-monophosphate-activated protein kinase-associated activation of autophagy, which in turn facilitates the removal of dysfunctional mitochondria and promotes survival of keratinocytes. Thus, we suggest that low-level colonization of hair follicles with noninvasive P. acnes strains, by triggering a local increase in autophagic activity, might exert a profound effect on several physiological processes responsible for the maintenance of skin tissue homeostasis.