Impaired nucleotide excision repair pathway as a possible factor in pathogenesis of head and neck cancer.

Tobacco smoking is one of the major risk factors in pathogenesis of head and neck squamous cell carcinomas (HNSCC). Many of the chemical compounds present in tobacco are well-known carcinogens which form adducts with DNA. Cells remove these adducts mainly by the nucleotide excision repair pathway (NER). NER also eliminates a broad spectrum of pyrimidine dimers (CPD) and photo-products (6-4PP) induced by UV-radiation or DNA cross-links after cisplatin anti-cancer treatment. In this study DNA damage and repair was examined in peripheral blood lymphocytes obtained from 20 HNSCC patients and 20 healthy controls as well as HTB-43 larynx and SSC-25 tongue cancer cell lines. DNA repair kinetics in the examined cells after cisplatin or UV-radiation treatment were investigated using alkaline comet assay during 240min of post-treatment incubation. MTT assay was used to analyse cell viability and the Annexin V-FITC kit specific for kinase-3 was employed to determine apoptosis after treating the cells with UV-radiation at dose range from 0.5 to 60J/m(2). NER capability was assessed in vitro with cell extracts by the use of a bacterial plasmid irradiated with UV-light as a substrate for the repair. The results show that lymphocytes from HNSCC patients and HTB-43 or SSC-25 cancer cells were more sensitive to genotoxic treatment with UV-radiation and displayed impaired DNA repair. Also evidenced was a higher rate of apoptosis induction after UV-radiation treatment of lymphocytes from the HNSCC patients and the HTB-43 cancer cells than after treatment of those from healthy donors. Finally, our results showed that there was a significant decrease in NER capacity in HTB-43 or SSC-25 cancer cells as well as in peripheral blood lymphocytes of HNSCC patients compared to controls. In conclusion, we suggest that the impaired NER pathway might be a critical factor in pathogenesis of head and neck cancer.

DNA double strand breaks repair and apoptosis induction in peripheral blood lymphocytes of head and neck cancer patients.

AIM: To evaluate the generation and repair of DNA double strand breaks (DSBs) as a critical factors that define the efficiency of radiation therapy of cancer patients. METHODS: Peripheral blood lymphocytes obtained from 18 patients with head and neck squamous cell carcinoma (HNSCC) and 18 healthy donors were studied. The efficiency of DSBs repair after genotoxic treatment with hydrogen peroxide and gamma-radiation were examined by neutral comet assay. MTT assay was used for cell viability analysis and Annexin V-FITC kit specific for kinase-3 was employed to determine apoptosis. RESULTS: Lymphocytes from HNSCC patients were sensitive to genotoxic treatment and displayed impaired DSBs repair. Finally, as a consequence of this finding we have evidenced higher rate of apoptosis induction after gamma-radiation treatment of lymphocytes from HNSCC patients than those from healthy controls. CONCLUSIONS: DSBs repair and increased apoptosis in cells of patients with head and neck cancer is relevant for efficient therapy of HNSCC.

Polymorphisms of the DNA base excision repair gene MUTYH in head and neck cancer.

BACKGROUND: Head and neck squamous cell carcinomas (HNSCC) comprise about 6% of all malignant neoplasms. The major risk factors of -HNSCC are smoking and alcohol consumption. Genetic polymorphisms of DNA repair enzymes may lead to genetic instability and carcinogenesis. MUTYH gene encodes a DNA glycosylase that can initiate the base excision repair (BER) pathway and prevent G:C > T:A transversion by excising adenine mispaired with 8-hydroxyguanine produced by reactive oxygen species (ROS). AIM: to perform a case-control study to test the association between polymorphism in the MUTYH gene: Tyr165Cys and head and neck cancer risk progression. METHODS: Genotypes were determined in DNA from peripheral blood lymphocytes of 193 patients (among them 97 subjects with precancerous hyperplastic laryngeal lesions and 96 subjects with head and neck cancer) and 140 age, sex and ethnic-matched cancer-free controls by tetra-primer amplification refractory mutation system PCR (T-ARMS-PCR). RESULTS: We found an association between head and neck cancer risk and the Tyr165Tyr variant of the MUTYH gene (OR 2.18; 95% CI 1.19-3.97). For Tyr165Tyr genotype we also observed positive correlation with cancer progression assessed by tumor size (OR 4.56; 95% CI 1.60-12.95). We did not observe any correlation between Tyr165Cys polymorphism of MUTYH gene and precancerous hyperplastic laryngeal lesions risk. CONCLUSION: The Tyr165Tyr polymorphic variant of the MUTYH gene may be associated with head and neck cancer in Polish population.

Tracking the concentration of heterotrophic plate count bacteria from the source to the consumer's tap.

The goal of this project was to quantify the concentration of heterotrophic plate count (HPC) bacteria within water reaching consumer's taps, and from the sources used by a major utility serving the City of Tucson, AZ. With this information, the amounts and sources of HPC bacteria consumed at the tap could be determined. Samples of water were collected on a monthly basis from two well fields, the CAVSARP recovery well field and Southern Avra Valley well field which serves as one of the groundwater sources for Tucson, AZ, and the distribution system which serves the same homes from which tap water was also tested. The average concentration of HPC in source waters within Southern Avra Valley Wells was 56 CFU/ml (range 1-1995/ml). From the CAVSARP recovery well field, corresponding values were 38 CFU/ml (1 to 502 CFU/ml). Unblended groundwater in the chlorinated distribution system averaged 22 CFU/ml (range 1-794). Blended water at the chlorinated distribution site averaged 47 CFU/ml (range 10-158). There was a major shift in the percentage of gram negative to gram-positive bacteria from the wells to the distribution system, to the tap. In the surface CAP source water, 76% of the bacteria were gram-negative compared to 27% gram-negative in the CAVSARP recovery wells. In contrast, Avra Valley wells contained 17% gram-negative bacteria. In both the Tucson groundwater distribution sites and blended distribution sites, the corresponding number of gram negative bacteria was 12%. Finally at the tap, only 0.2% of the bacteria were gram-negative. The average number of bacteria in household taps averaged 3072 HPC/ml and was equal or greater than 500 ml 68% of the time. This study shows that the number of HPC bacteria increases dramatically from the distribution system to the consumers tap. Thus, the major source of bacteria ingested by the average consumer in Tucson originates from bacteria within the household distribution system or the household tap, rather than from source waters or the distribution system. It is also clear that consumers' regularly consume more than 500 HPC/ml from drinking water taken from the household tap.

Rapid reduction of Legionella pneumophila on stainless steel with zeolite coatings containing silver and zinc ions.

AIMS: To determine the rate of reduction of Legionella pneumophila by stainless steel surfaces with zeolite ceramic coatings containing 2.5% (w/w) silver (Ag) and 14% zinc (Zn) ions. METHODS AND RESULTS: Stainless steel pans with and without Ag/Zn coatings were inoculated with solutions of Leg. pneumophila ATCC 33155 and incubated at 37 degrees C. Survival was monitored using the spread-plate technique on selective buffered charcoal yeast extract agar. Significant reductions of Leg. pneumophila were effected by the Ag/Zn zeolite coatings within 2 h of exposure. CONCLUSIONS, SIGNIFICANCE AND IMPACT OF THE STUDY: Zeolite ceramic Ag/Zn coatings impart significant anti-Legionella properties to stainless steel surfaces. Coated stainless steel could be used in the manufacture of air ducts, condensation pans and intake and exhaust vents. These products have the potential to reduce numbers of Legionella in air-handling systems.

Rapid reduction of Staphylococcus aureus populations on stainless steel surfaces by zeolite ceramic coatings containing silver and zinc ions.

This study demonstrates the anti-Staphylococcus aureus properties of stainless steel surfaces coated with zeolite containing 2.5% silver and 14% zinc ions. Stainless steel panels with and without the heavy-metal-containing coatings were inoculated with S. aureus and incubated at room temperature. Survival of S. aureus was significantly reduced by the silver/zinc coatings within 1 h. Many hospital surfaces could be constructed of stainless steel with silver/zinc zeolite coatings. Such measures may reduce rates of hospital-acquired S. aureus infection.

Quantitative assessment of risk reduction from hand washing with antibacterial soaps.

The Centers for Disease Control and Prevention have estimated that there are 3,713,000 cases of infectious disease associated with day care facilities each year. The objective of this study was to examine the risk reduction achieved from using different soap formulations after diaper changing using a microbial quantitative risk assessment approach. To achieve this, a probability of infection model and an exposure assessment based on micro-organism transfer were used to evaluate the efficacy of different soap formulations in reducing the probability of disease following hand contact with an enteric pathogen. Based on this model, it was determined that the probability of infection ranged from 24/100 to 91/100 for those changing diapers of babies with symptomatic shigellosis who used a control product (soap without an antibacterial ingredient), 22/100 to 91/100 for those who used an antibacterial soap (chlorohexadine 4%), and 15/100 to 90/100 for those who used a triclosan (1.5%) antibacterial soap. Those with asymptomatic shigellosis who used a non-antibacterial control soap had a risk between 49/100,000 and 53/100, those who used the 4% chlorohexadine-containing soap had a risk between 43/100,000 and 51/100, and for those who used a 1.5% triclosan soap had a risk between 21/100,000 and 43/100. The adequate washing of hands after diapering reduces risk and can be further reduced by a factor of 20% by the use of an antibacterial soap. Quantitative risk assessment is a valuable tool in the evaluation of household sanitizing agents and low risk outcomes.

Comparative surface-to-hand and fingertip-to-mouth transfer efficiency of gram-positive bacteria, gram-negative bacteria, and phage.

AIMS: To determine the transfer efficiency of micro-organisms from fomites to hands and the subsequent transfer from the fingertip to the lip. METHODS AND RESULTS: Volunteers hands were sampled after the normal usage of fomites seeded with a pooled culture of a Gram-positive bacterium (Micrococcus luteus), a Gram-negative bacterium (Serratia rubidea) and phage PRD-1 (Period A). Activities included wringing out a dishcloth/sponge, turning on/off a kitchen faucet, cutting up a carrot, making hamburger patties, holding a phone receiver, and removing laundry from the washing machine. Transfer efficiencies were 38.47% to 65.80% and 27.59% to 40.03% for the phone receiver and faucet, respectively. Transfer efficiencies from porous fomites were <0.01%. In most cases, M.luteus was transferred most efficiently, followed by phage PRD-1 and S. rubidea. When the volunteers' fingertips were inoculated with the pooled organisms and held to the lip area (Period B), transfer rates of 40.99%, 33.97%, and 33.90% occurred with M. luteus, S. rubidea, and PRD-1, respectively. CONCLUSIONS: The highest bacteral transfer rates from fomites to the hands were seen with the hard, non-porous surfaces. Even with low transfer rates, the numbers of bacteria transferred to the hands were still high (up to 10(6) cells). Transfer of bacteria from the fingertip to the lip is similar to that observed from hard surfaces to hands. SIGNIFICANCE AND IMPACT OF THE STUDY: Infectious doses of pathogens may be transferred to the mouth after handling an everyday contaminated household object.

Quantitative assessment of risk reduction from hand washing with antibacterial soaps.

The Centers for Disease Control and Prevention have estimated that there are 3,713,000 cases of infectious disease associated with day care facilities each year. The objective of this study was to examine the risk reduction achieved from using different soap formulations after diaper changing using a microbial quantitative risk assessment approach. To achieve this, a probability of infection model and an exposure assessment based on micro-organism transfer were used to evaluate the efficacy of different soap formulations in reducing the probability of disease following hand contact with an enteric pathogen. Based on this model, it was determined that the probability of infection ranged from 24/100 to 91/100 for those changing diapers of babies with symptomatic shigellosis who used a control product (soap without an antibacterial ingredient), 22/100 to 91/100 for those who used an antibacterial soap (chlorohexadine 4%), and 15/100 to 90/100 for those who used a triclosan (1.5%) antibacterial soap. Those with asymptomatic shigellosis who used a non-antibacterial control soap had a risk between 49/100,000 and 53/100, those who used the 4% chlorohexadine-containing soap had a risk between 43/100,000 and 51/100, and for those who used a 1.5% triclosan soap had a risk between 21/100,000 and 43/100. The adequate washing of hands after diapering reduces risk and can be further reduced by a factor of 20% by the use of an antibacterial soap. Quantitative risk assessment is a valuable tool in the evaluation of household sanitizing agents and low risk outcomes.

Reduction of faecal coliform, coliform and heterotrophic plate count bacteria in the household kitchen and bathroom by disinfection with hypochlorite cleaners.

Fourteen sites evenly divided between the household kitchen and bathroom were monitored on a weekly basis for numbers of faecal coliforms, total coliforms and heterotrophic plate count bacteria. The first 10 weeks comprised the control period, hypochlorite cleaning products were introduced into the household during the second 10 weeks, and a strict cleaning regimen using hypochlorite products was implemented during the last 10 weeks. The kitchen was more heavily contaminated than the bathroom, with the toilet seat being the least contaminated site. The highest concentrations of all three classes of bacteria were found on sites that were moist environments and/or were frequently touched; these included the sponge/dishcloth, the kitchen sink drain area, the bath sink drain area, and the kitchen faucet handle(s). The implementation of a cleaning regimen with common household hypochlorite products resulted in the significant reduction of all three classes of bacteria at these four sites and other household sites.

Risk assessment of opportunistic bacterial pathogens in drinking water.

This study was undertaken to examine quantitatively the risks to human health posed by heterotrophic plate count (HPC) bacteria found naturally in ambient and potable waters. There is no clear-cut evidence that the HPC bacteria as a whole pose a public health risk. Only certain members are opportunistic pathogens. Using the four-tiered approach for risk assessment from the National Academy of Sciences, hazard identification, dose-response modeling, and exposure through ingestion of drinking water were evaluated to develop a risk characterization, which estimates the probability of infection for individuals consuming various levels of specific HPC bacteria. HPC bacteria in drinking water often include isolates from the following genera: Pseudomonas, Acinetobacter, Moraxella, Aeromonas, and Xanthomonas. Other bacteria that are commonly found are Legionella and Mycobacterium. All these genera contain species that are opportunistic pathogens which may cause serious diseases. For example, the three nonfermentative gram-negative rods most frequently isolated in the clinical laboratory are (1) Pseudomonas aeruginosa, (2) Acinetobacter, and (3) Xanthomonas maltophilia. P. aeruginosa is a major cause of hospital-acquired infections with a high mortality rate. Aeromonas is sometimes associated with wound infections and suspected to be a causative agent of diarrhea. Legionella pneumophila causes 4%-20% of cases of community-acquired pneumonia and has been ranked as the second or third most frequent cause of pneumonia requiring hospitalization. The number of cases of pulmonary disease associated with Mycobacterium avian is rapidly increasing and is approaching the incidence of M. tuberculosis in some areas. Moraxella can cause infections of the eye and upper respiratory tract. The oral infectious doses are as follows in animal and human test subjects: P. aeruginosa, 10(8)-10(9); A, hydrophila, > 10(10); M. avium, 10(4)-10(7); and X. maltophilia, 10(6)-10(9). The infectious dose for an opportunistic pathogen is lower for immunocompromised subjects or those on antibiotic treatment. These bacteria have been found in drinking water at the following frequencies: P. aeruginosa, < 1%-24%; Acinetobacter, 5%-38%; X. maltophilia, < 1%-2%; Aeromonas, 1%-27%; Moraxella, 10%-80%; M. avium, < 1%-50%; and L. pneumophila, 3%-33%. These data suggest that drinking water could be a source of infection for some of these bacteria. The risk characterization showed that risks of infection from oral ingestion ranged from a low of 7.3 x 10(-9) (7.3/billion) for low exposures to Aeromonas to higher risks predicted at high levels of exposure to Pseudomonas of 9 x 10(-2) (98/100). This higher risk was only predicted for individuals on antibiotics. Overall, the evidence suggests that specific members of HPC bacteria found in drinking water may be causative agents of both hospital- and community-acquired infections. However, the case numbers may be very low and the risks represent levels generally less than 1/10,000 for a single exposure to the bacterial agent. Future research needs include (1) determining the seasonal concentrations of these bacteria in drinking water, (2) conducting adequate dose-response studies in animal subjects or human volunteers, (3) determining the health risks for an individual with multiple exposures to the opportunistic pathogens, and (4) evaluating the increase in host susceptibility conferred by antibiotic use or immunosuppression.

Haemophilus influenzae: an important cause of maternal and neonatal infections.

Although Haemophilus influenzae is recognized as a major pathogen of infants, its role in maternal and neonatal infections is not as well appreciated. We analyzed the records of all mothers and neonates infected with H influenzae over a 10-year period. Twenty-eight mother/neonate sets were identified in which at least one had documented infection with H influenzae. Of the 18 mothers with documented infection, 13 had chorioamnionitis, endometritis, or both, and two of these mothers were bacteremic with H influenzae. Of the 23 infected neonates, 15 presented with early sepsis and/or pneumonia and nine had conjunctivitis. During the period of the study, only group B streptococci and Escherichia coli were more common as causes of early neonatal bacteremia. Under the conditions of this retrospective study, maternal infection predicted neonatal infection. However, prospective studies in which asymptomatic patients are cultured will be required to determine how well maternal colonization/infection with H influenzae predicts neonatal infection.