Cytogenetic analysis of buccal cells from shoeworkers and pathology and anatomy laboratory workers exposed to n-hexane,toluene, methyl ethyl ketone and formaldehyde

People employed in the shoe manufacture and repair industry are at an increased risk for cancer, the strongest evidence being for nasal cancer and leukaemia. A possible causal role for formaldehyde is likely for cancer of the buccal cavity and nasopharynx. Exfoliated buccal cells are good source of tissue for monitoring human exposure to inhaled and ingested occupational and environmental genotoxicants. To assess the cytogenetic damage related to occupational exposure to airborne chemicals during shoe-making and the processes in pathology and anatomy laboratories, the micronuclei (MN) count per 3000 cells was measured in buccal smears from shoe-workers (group I, n = 22) exposed to mainly n-hexane, toluene and methyl ethyl ketone (MEK) and from anatomy and pathology staff (group II, n = 28) exposed to formaldehyde (FA). Eighteen male university staff were used as controls. The mean time-weighted average (TWA) concentrations of n-hexane, toluene and MEK in 10 small shoe workshops were 58.07 p.p.m., 26.62 p.p.m. and 11.39 p.p.m., respectively. The measured air concentrations of FA in the breathing zone of the anatomy and pathology laboratory workers were between 2 and 4 p.p.m. Levels of 2,5-hexadione (2,5-HD) and hippuric acid (HA), metabolic markers of n-hexane and toluene exposure, respectively, were significantly higher in the urine of workers in group I than in control subjects (p < 0.001 and p < 0.01, respectively). The mean (±SD) MN frequencies in buccal mucosa cells from workers in group I, group II and controls were 0.62±0.45%, 0.71±0.56% and 0.33±0.30%, respectively (p < 0.05 and p < 0.05 compared with controls for group I and group II, respectively). The effects of smoking, age and duration of exposure on the frequency of micronucleated buccal cells from workers in all three groups studied were also evaluated. Overall, the results suggest that occupational exposure to organic solvents, mainly n-hexane, toluene, MEK and FA, may cause cytogenetic damage in buccal cells and that use of exfoliated buccal cells seems to be appropriate to measure exposure to organic solvents.     

Resveratrol Induced Premature Senescence Is Associated with DNA Damage Mediated SIRT1 and SIRT2 Down-Regulation

The natural polyphenolic compound resveratrol (3,4,5-trihydroxy-trans-stilbene) has broad spectrum health beneficial activities including antioxidant, anti-inflammatory, anti-aging, anti-cancer, cardioprotective, and neuroprotective effects. Remarkably, resveratrol also induces apoptosis and cellular senescence in primary and cancer cells. Resveratrol’s anti-aging effects both in vitro and in vivo attributed to activation of a (NAD)-dependent histone deacetylase family member sirtuin-1 (SIRT1) protein. In mammals seven members (SIRT1-7) of sirtuin family have been identified. Among those, SIRT1 is the most extensively studied with perceptive effects on mammalian physiology and suppression of the diseases of aging. Yet no data has specified the role of sirtuins, under conditions where resveratrol treatment induces senescence. Current study was undertaken to investigate the effects of resveratrol in human primary dermal fibroblasts (BJ) and to clarify the role of sirtuin family members in particular SIRT1 and SIRT2 that are known to be involved in cellular stress responses and cell cycle, respectively. Here, we show that resveratrol decreases proliferation of BJ cells in a time and dose dependent manner. In addition the increase in senescence associated β-galactosidase (SA-β-gal) activity and methylated H3K9-me indicate the induction of premature senescence. A significant increase in phosphorylation of γ-H2AX, a surrogate of DNA double strand breaks, as well as in levels of p53, p21^CIP1 and p16^INK4A is also detected. Interestingly, at concentrations where resveratrol induced premature senescence we show a significant decrease in SIRT1 and SIRT2 levels by Western Blot and quantitative RT-PCR analysis. Conversely inhibition of SIRT1 and SIRT2 via siRNA or sirtinol treatment also induced senescence in BJ fibroblasts associated with increased SA-β-gal activity, γ-H2AX phosphorylation and p53, p21^CIP1 and p16^INK4A levels. Interestingly DNA damaging agent doxorubicin also induced senescence in BJ fibroblasts associated with decreased SIRT1/2 levels. In conclusion our data reveal that resveratrol induced premature senescence is associated with SIRT1 and SIRT2 down regulation in human dermal fibroblasts. Here we suggest that the concomitant decline in SIRT1/2 expression in response to resveratrol treatment may be a cause for induction of senescence, which is most likely mediated by a regulatory mechanism activated by DNA damage response.     

Diversity and metabolic potential of culturable bacteria from the rhizosphere of Turkish tea grown in acidic soils

The purpose of this study was to investigate the diversity of cultivable phosphate solubilising (PSB) and total bacteria originated from 384 rhizospheric acidic soils samples of tea plants grown at 32 locations. Over 900 rhizoplane bacteria were randomly selected from agar-solidified trypticase soy broth, and identified using fatty acid methyl ester (FAME) profiles. Based on FAME profiles, 53 bacterial genera were identified with a similarity index >0.3, but 60.3% of the identified isolates belonged to five genera: Bacillus (34.6%), Pseudomonas (8.9%), Stenotrophomonas (6.1%), Paenibacillus (5.9%) and Arthrobacter (4.8%). The bacilli group comprised many different species, with the most abundant being B. cereus, B. megaterium and B. sphaericus. The main identified Pseudomonads included P. fluorescens, P. putida, and P. alcaligenes. About 30.4% of the bacterial isolates could not be classified to genus since their similarity indices were <0.3 indicating no close matches. Most of the total and P-solubilizing bacteria isolated were Gram positive (61.3 and 52.3%), and Gram negative constituted only 38.7 and 47.7%. Out of the 214 PSB from a pool of 506 bacterial isolates recovered on the selective media from the rhizosphere of tea, 74 of them were characterized by carbon sources using BIOLOG^M GN2 and GP2 plates. Bacillus, Pseudomonas, Paenibacillus and Stenotrophomonas genera were the most prominent P-solubilizing groups in the rhizosphere and soil populations analyzed. B. cereus, P. fluorescens, S. maltophilia, B. megaterium, P. putida, B. sphaericus and Paenibacillus polymyxa were the most frequent P-solubilizing species in the acidic tea rhizosohere soils. Selected Gram-positive PSB appeared to favour carbohydrates, and Gram-negative bacteria appeared to favour carboxylic acids, amino acids and carbohydrates as carbon sources. Selected phosphate solubilizing acid tolerant strains showed high variability in utilizing various carbon sources.     

Assessment of the risk of perforation of the mandibular canal by implant drill using density and thickness parameters

Gerodontology 2009; doi: 10.1111/j.1741‐2358.2009.00362.x
Assessment of the risk of perforation of the mandibular canal by implant drill using density and thickness parameters Objective: The objective of this study was to investigate whether the resistance of the bone surrounding the mandibular canal had sufficient density and thickness to avoid perforation by drills when preparing the bed of the implant. Background: Damage to the inferior alveolar nerve (IAN) is more common than expected. This injury may lead to serious complications ranging from mild paresthesia to total anaesthesia of the lower jaw. Materials and methods: The CT images of 99 patients, whose ages ranged between 20 and 79 years, and who applied for an implant application to the posterior aspect of the mandible were included in this study. Results: The overall average bone thickness in the premolar and molar regions was 0.8717 ± 0.1818 and 0.8556 ± 0.1756 mm, respectively, whereas the bone density in the premolar and molar regions was 649.18 ± 241.42 and 584.44 ± 222.73 Hounsfield Units (HU), respectively (p < 0.001). Conclusion: It was determined that the average density and thickness of the bone that surrounds the mandibular canal was not sufficient to resist the implant drill. It can be concluded that the risk of injury to the IAN may be minimised by accurately determining the bone mass on the canal prior to the implant procedure, and avoiding excessive force when approaching the canal.     

Sensory projections to the nucleus basalis prosencephali of the pigeon

Summary The afferent pathways to the nucleus basalis prosencephali of the pigeon were studied by use of the horseradish peroxidase (HRP) technique. It was confirmed that this nucleus receives a direct pathway from the nucleus sensorius principalis nervi trigemini and that, as in the starling, it receives a direct input from the nucleus lemnisci lateralis, pars ventralis, an auditory relay. Totally novel is the finding that the nucleus basalis prosencephali is the target of a direct pathway originating in the medullary nucleus vestibularis superior. All three pathways bypass the thalamus. From within the telencephalon the nucleus basalis prosencephali also receives fibres from the tuberculum olfactorium and the peri-ectostriatal belt, suggestive of olfactory and visual input. Marked cell bodies were also found in the neostriatum frontolaterale. It is assumed that these arose from HRP uptake by axons of the tractus fronto-archistriatalis that course through the nucleus basalis prosencephali to the anterodorsal archistriatum. Marked fibres and bouton-like formations were observed in the latter structure. The afferents to the nucleus basalis prosencephali are discussed in conjunction with the probable role of the nucleus as a sensorimotor coordinator of the pecking/feeding behaviour of the pigeon.     

Necroptosis Interfaces with MOMP and the MPTP in Mediating Cell Death

During apoptosis the pro-death Bcl-2 family members Bax and Bak induce mitochondrial outer membrane permeabilization (MOMP) to mediate cell death. Recently, it was shown that Bax and Bak are also required for mitochondrial permeability transition pore (MPTP)-dependent necrosis, where, in their non-oligomeric state, they enhance permeability characteristics of the outer mitochondrial membrane. Necroptosis is another form of regulated necrosis involving the death receptors and receptor interacting protein kinases (RIP proteins, by Ripk genes). Here, we show cells or mice deficient for Bax/Bak or cyclophilin D, a protein that regulates MPTP opening, are resistant to cell death induced by necroptotic mediators. We show that Bax/Bak oligomerization is required for necroptotic cell death and that this oligomerization reinforces MPTP opening. Mechanistically, we observe mixed lineage kinase domain-like (MLKL) protein and cofilin-1 translocation to the mitochondria following necroptosis induction, while expression of the mitochondrial matrix isoform of the antiapoptotic Bcl-2 family member, myeloid cell leukemia 1 (Mcl-1), is significantly reduced. Some of these effects are lost with necroptosis inhibition in Bax/Bak1 double null, Ppif^-/-, or Ripk3^-/- fibroblasts. Hence, downstream mechanisms of cell death induced by necroptotic stimuli utilize both Bax/Bak to generate apoptotic pores in the outer mitochondrial membrane as well as MPTP opening in association with known mitochondrial death modifying proteins.     

Investigation of C5a receptor gene 450 C/T polymorphism in Turkish patients with familial Mediterranean fever

Familial Mediterranean fever (FMF) is a genetic disorder with acute inflammatory serosal attacks due to MEFV gene mutations which resides in chromosome 16. Lack of a C5a inhibitor activity in the peritoneum has previously been proposed in part to contribute in propagation of the serosal inflammation in FMF attacks. The aim of this study is to investigate C5a receptor (C5aR) gene polymorphism in patients with FMF and its relation to the main features of the disease. A polymorphism in the coding region of C5aR gene leading to C to T transition at nucleotide position 450 has been investigated in 85 non-related Turkish FMF patients and 160 non-related healthy controls by using PCR-RFLP. The frequencies of C5aR gene 450 CT genotype and T allele were not significantly different between Turkish FMF patients and healthy subjects (14.12 and 8.24% for FMF vs. 10 and 5% for controls, respectively). C5aR gene 450 CT genotype tended to associate with the presence Henoch-Schonlein purpura (OR: 1.25, 95% CI: 0.917–1.704, P = 0.017) but with no other clinical findings of the disease. C5aR polymorphism might be searched in populations having high prevalence of FMF.     

Temperature-Dependent Requirement for Catalase in Aerobic Growth of Listeria monocytogenes F2365

Listeria monocytogenes is a Gram-positive, psychrotrophic, facultative intracellular food-borne pathogen responsible for severe illness (listeriosis). The bacteria can grow in a wide range of temperatures (1 to 45°C), and low-temperature growth contributes to the food safety hazards associated with contamination of ready-to-eat foods with this pathogen. To assess the impact of oxidative stress responses on the ability of L. monocytogenes to grow at low temperatures and to tolerate repeated freeze-thaw stress (cryotolerance), we generated and characterized a catalase-deficient mutant of L. monocytogenes F2365 harboring a mariner-based transposon insertion in the catalase gene (kat). When grown aerobically on blood-free solid medium, the kat mutant exhibited impaired growth, with the extent of impairment increasing with decreasing temperature, and no growth was detected at 4°C. Aerobic growth in liquid was impaired at 4°C, especially under aeration, but not at higher temperatures (10, 25, or 37°C). Genetic complementation of the mutant with the intact kat restored normal growth, confirming that inactivation of this gene was responsible for the growth impairment. In spite of the expected impact of oxidative stress responses on cryotolerance, cryotolerance of the kat mutant was not affected.Listeria monocytogenes is a Gram-positive, facultative intracellular food-borne pathogen that has the ability to cause a severe disease (listeriosis) in humans and animals (13, 28, 30). L. monocytogenes is ubiquitously distributed in the environment and has the ability to grow over a wide range of temperatures (between 1 and 45°C) (13). Growth at low temperature has important implications for environmental persistence of the organism and for contamination of cold-stored, ready-to-eat foods, thus contributing to the food safety hazards associated with L. monocytogenes (19).L. monocytogenes is subjected to oxidative stress during both extracellular and intracellular growth and has evolved several responses to minimize the impact of reactive oxygen species (ROS). Catalase and superoxide dismutase (SOD) work synergistically in detoxification of ROS: superoxide anions are converted to H₂O₂ by SOD, with subsequent conversion of H₂O₂ into water and oxygen by catalase (22). Exposure to ROS may be especially acute during intracellular infection as well as under certain environmental conditions, such as those involved in repeated freezing and thawing (15, 16, 23, 29, 33).Previous studies revealed that the ability of L. monocytogenes to survive repeated freezing and thawing (cryotolerance) was markedly dependent on growth temperature, with bacteria grown at 37°C having significantly higher cryotolerance than those grown at either 4 or 25°C (1). However, mechanisms underlying Listeria''s cryotolerance have not been identified. Since oxidative damage is considered to take place during freezing and thawing, determinants such as catalase may be involved in cryotolerance.The catalase of L. monocytogenes has been investigated primarily in terms of its potential role in pathogenesis, with somewhat conflicting results. The isolation of catalase-negative strains from human listeriosis patients has led to the speculation that catalase is not required for human virulence (4, 8, 12, 31). On the other hand, under certain conditions (e.g., reduced serum levels), catalase-negative strains were impaired in their ability to survive in activated macrophages in comparison to catalase-positive strains (32). Furthermore, the catalase gene kat was among those for which expression was induced in infected cell cultures and in the spleens of mice infected with L. monocytogenes EGD-e, suggesting possible contributions to pathogenesis (5, 9).The potential role of catalase in environmental adaptations of L. monocytogenes such as growth at low temperature and cryotolerance was not addressed in these earlier investigations. In this study, we have characterized an isogenic mutant of L. monocytogenes F2365 to determine the involvement of catalase in growth at different temperatures, survival in selected foods, and cryotolerance of L. monocytogenes.