The impact of bioaerosol trajectories on microbial community assembly and physicochemical dynamics in the atmosphere.

This study explored the assembly mechanisms and physicochemical dynamics of microbial communities within atmospheric bioaerosols, focusing on the influence of different aerial trajectories. Over two years, samples near Seoul were classified into 'North', 'Southwest', and 'Others' categories based on their aerial trajectories. Physicochemical analysis of the PM2.5 particles revealed distinct ion compositions for each cluster, reflecting diverse environmental influences. Microbial community analysis revealed that shared dominant bacterial phyla were present in all clusters. However, distinct taxonomic profiles and biomarkers were also evident, such as coastal bacteria in the 'Southwest' cluster correlating with wind speed, and arid soil-originated bacteria in the 'North' cluster correlating with cations. These findings demonstrate that biomarkers in each cluster are representative of the distinct environments associated with their aerial trajectories. Notably, cluster 'Southwest' the highest microbial diversity and a strong alignment with the neutral community model, suggesting a large influence of passive dispersal from marine environments. Contrarily, 'North' and 'Others' were more influenced by niche-dependent factors. This study highlights the complex interplay between environmental factors and microbial dynamics in bioaerosols and provides important insights for environmental monitoring and public health risk assessment.

Langerhans Cell Histiocytosis in the Nasal Bone: A Rare Case.

Nasal bone involvement of Langerhans cell histiocytosis is rarely reported. Here we present a case of a 13-year-old boy with a palpable nasal mass. Ultrasonography revealed a hypoechoic mass on the left side of the nose. Both CT scanning and MRI showed an osteolytic mass. The lesion seen on MRI was well-defined mass with homogeneous enhancement. Histopathological examination of the resected specimen confirmed the diagnosis of LCH.

90-day nose-only inhalation toxicity study of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) in Sprague-Dawley rats.

4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) is one of the key tobacco-specific nitrosamines that plays an important role in human lung carcinogenesis. Repeated dose inhalation toxicity data on NNK, particularly relevant to cigarette smoking, however, is surprisingly limited. Hence, there is a lack of direct information available on the carcinogenic and potential non-carcinogenic effects of NNK via inhalational route exposure. In the present study, the subchronic inhalation toxicity of NNK was evaluated in Sprague Dawley rats. Both sexes (9-10 weeks age; 23 rats/sex/group) were exposed by nose-only inhalation to air, vehicle control (75% propylene glycol), or 0.2, 0.8, 3.2, or 7.8 mg/kg body weight (BW)/day of NNK (NNK aerosol concentrations: 0, 0, 0.0066, 0.026, 0.11, or 0.26 mg/L air) for 1 h/day for 90 consecutive days. Toxicity was evaluated by assessing body weights; food consumption; clinical pathology; histopathology; organ weights; blood, urine, and tissue levels of NNK, its major metabolite 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL), and their glucuronides (reported as total NNK, tNNK, and total NNAL, tNNAL, respectively); tissue levels of the DNA adduct O6-methylguanine; blood and bone marrow micronucleus (MN) frequency; and bone marrow DNA strand breaks (comet assay). The results showed that NNK exposure caused multiple significant adverse effects, with the most sensitive endpoint being non-neoplastic lesions in the nose. Although the genotoxic biomarker O6-methylguanine was detected, genotoxicity from NNK exposure was negative in the MN and comet assays. The Lowest-Observed-Adverse-Effect-Level (LOAEL) was 0.8 mg/kg BW/day or 0.026 mg/L air of NNK for 1 h/day for both sexes. The No-Observed-Adverse-Effect-Level (NOAEL) was 0.2 mg/kg BW/day or 0.0066 mg/L air of NNK for 1 h/day for both sexes. The results of this study provide new information relevant to assessing the human exposure hazard of NNK.

14-Day Nose-Only Inhalation Toxicity and Haber's Rule Study of NNK in Sprague-Dawley Rats.

4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) is one of the key tobacco-specific nitrosamines that plays an important role in human lung carcinogenesis. However, repeated inhalation toxicity data on NNK, which is more directly relevant to cigarette smoking, are currently limited. In the present study, the subacute inhalation toxicity of NNK was evaluated in Sprague Dawley rats. Both sexes (9-10 weeks age; 16 rats/sex/group) were exposed by nose-only inhalation to air, vehicle control (75% propylene glycol), or 0.8, 3.2, 12.5, or 50 mg/kg body weight (BW)/day of NNK (NNK aerosol concentrations: 0, 0, 0.03, 0.11, 0.41, or 1.65 mg/L air) for 1 h/day for 14 consecutive days. Toxicity was evaluated by assessing body and organ weights; food consumption; clinical pathology; histopathology observations; blood, urine, and tissue levels of NNK, its major metabolite 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL), and their glucuronides (reported as total NNK, tNNK, and total NNAL, tNNAL, respectively); O6-methylguanine DNA adduct formation; and blood and bone marrow micronucleus frequency. Whether the subacute inhalation toxicity of NNK followed Haber's Rule was also determined using additional animals exposed 4 h/day. The results showed that NNK exposure caused multiple significant adverse effects, with the most sensitive endpoint being non-neoplastic histopathological lesions in the nose. The lowest-observed-adverse-effect level (LOAEL) was 0.8 mg/kg BW/day or 0.03 mg/L air for 1 h/day for both sexes. An assessment of Haber's Rule indicated that 14-day inhalation exposure to the same dose at a lower concentration of NNK aerosol for a longer time (4 h daily) resulted in greater adverse effects than exposure to a higher concentration of NNK aerosol for a shorter time (1 h daily).

Sequential biowashing-biopile processes for remediation of crude oil contaminated soil in Kuwait.

The application of biological processes for remediation of the aged crude oil-contaminated soil of Kuwait can be an inefficient way, thus, this study developed 20 d-sequential biowashing and biopile processes where the biowashing step uses an enrichment culture of the indigenous soil bacterial community and the biopile step includes hemoglobin-catalyzed oxidation (HCO). The residual total petroleum hydrocarbons (TPH) concentrations and CO2 generation were measured to determine the removal efficiency, and the bacterial community changes were studied to investigate the effect of the sequential processes on the soil indigenous bacterial community. The enrichment culture grown on hemoglobin showed an increased surface activity, and this promoted desorption and emulsification of crude oil from the soil sample in the biowashing step resulting in 75% TPH removal. Potential surfactant-producing bacterial species were observed in the soil sample after biowashing. The HCO in the beginning of the biopile step removed 21% of the residual TPH, and further TPH removal was observed with a longer biopile period. Overall, the sequential biowashing and biopile processes removed 86% TPH. The results show that the developed sequential biowashing and biopile processes can be used to efficiently remediate the aged crude oil-contaminated soil of Kuwait.

Evaluation of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) mutagenicity using in vitro and in vivo Pig-a assays.

4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) is a genotoxic carcinogen found in tobacco and tobacco smoke. Several in vitro and in vivo assays have been used for evaluating the genotoxicity of tobacco smoke and tobacco smoke constituents like NNK, yet it is not clear which in vitro assays are most appropriate for extrapolating the in vitro responses of these test agents to animal models and humans. The Pig-a gene mutation assay can be performed in vitro, in laboratory animals, and in humans, a potential benefit in estimating in vivo responses from in vitro data. In the current study we used Pig-a as a reporter of gene mutation both in vitro, in L5178Y/Tk+/- cells, and in vivo, in Sprague-Dawley rats. NNK significantly increased Pig-a mutant frequency in L5178Y/Tk+/- cells, but only at concentrations of 100 µg/ml and greater, and only in the presence of S9 activation. Pig-a mutations in L5178Y/Tk+/- cells were detected in 80% of the NNK-induced mutants, with the predominate mutation being G→A transition; vehicle control mutants contained deletions. In the in vivo study, rats were exposed to NNK daily for 90 days by inhalation, a common route of exposure to NNK for humans. Although elevated mutant frequencies were detected, these responses were not clearly associated with NNK exposure, so that overall, the in vivo Pig-a assays were negative. Thus, while NNK induces mutations in the in vitro Pig-a assay, the in vivo Pig-a assay has limited ability to detect NNK mutagenicity under conditions relevant to NNK exposure in smokers.

Pedobacter solisilvae sp. nov., isolated from forest soil.

The bacteria strain EN12T was isolated from forest soil in the Republic of Korea. The cells were Gram-negative, non-motile and rod-shaped, and the strain was strictly aerobic. Phylogenetic analysis of its 16S rRNA gene sequences showed that strain EN12T belonged to the class Sphingobacteriia of the phylum Bacteroidetes, and its closest relative is Pedobacter namyangjuensis 5G38T, with a sequence similarity of 95.5 %. The average DNA sequence similarity from validly described species within the genus Pedobacter was 92.5±1.3 %. Chemotaxonomic data including major ubiquinones (menaquinone-7), polar lipids (phosphatidylethanolamine and sphingolipid) and fatty acids (iso-C15 : 0, iso-C17 : 0 3-OH, and C16 : 1ω6c/C16 : 1ω7c) also supported an affiliation of strain EN12T with the genus Pedobacter. Genotypic and phenotypic differentiation of strain EN12T from six published Pedobacter species was revealed through DNA-DNA relatedness and physiological/biochemical tests. Results of these phenotypic, phylogenetic and chemotaxonomic analyses indicated that strain EN12T is a novel species in the genus Pedobacter, for which we propose the name Pedobacter solisilvae sp. nov. (=KCTC 42612T=LMG 28820T).

Chitinimonas lacunae sp. nov., isolated from artificial pond in Korea.

A gram-negative, rod shaped bacterium designated as strain H2T was isolated from an artificial pond in Korea. The strain H2T was able to grow aerobically and anaerobically with optimal growth occurring at 30 °C and pH 7.0 under aerobic conditions. Phylogenetic analysis based on 16S rRNA gene sequences showed that the strain H2T belonged to the genus Chitinimonas of the family Burkholderiaceae. Phylogenetic similarity calculated from 16S rRNA gene sequences of strain H2T and valid species belongs to the genus Chitinimonas ranged from 93.2 % (for Chitinimonas taiwanensis cfT) to 94.4 % (for Chitinimonas prasina LY03T), and strain H2T formed a tight monophyletic group with them. Predominant fatty acids were C16 : 0 and summed feature 3, which consisted of C16 : 1ω6c and/or C16 : 1ω7c. The major respiratory quinone of the strain H2T was ubiquinone-8, and DNA G+C content was 60.2 %. The polar lipids consisted of phosphatidylethanolamine, phosphatidylglycerol, unidentified aminolipid, and unidentified phospholipid. The biochemical characteristics that distinguished strain H2T from other Chitinimonas species included positive cystine arylamidase activity and lacked α-chymotrypsin and ß-glucosidase (aesculin hydrolysis) activity. In addition, reciprocal DNA-DNA relatedness between H2T and three Chitinimonas strains ranged from 32.0 to 43.7 %. On the basis of its phylogenetic, chemotaxonomic, and genotypic characteristics, strain H2T represents a novel species of the genus Chitinimonas. Chitinimonas lacunae sp. nov. is proposed with the type strain H2T (=KCTC 52574T=LMG 29894T).

Characteristics of orbital wall fractures in preschool and school-aged children.

OBJECTIVE: This study aimed to evaluate the injury patterns in pediatric patients with an orbital wall fracture (OWF) and to identify the differences in injury patterns between preschool and school-aged patients with OWF who presented to the emergency department. METHODS: We performed a retrospective observational study in the emergency department of a tertiary hospital between January 2004 and March 2014. A total of 177 pediatric patients (<18 years) with OWF who underwent facial bone computed tomography scans with specific discharge codes were included. Patients were categorized into preschool (≤7 years) and school-aged (>7 years) pediatric groups. RESULTS: The inferior wall was the most common fracture site in both the preschool and school-aged pediatric groups (50.0% vs. 64.4%, P=0.15). The male-to-female ratio and the mechanism of injury showed significant differences between the two age groups. Violence was the most common mechanism of injury in the school-aged pediatric group (49.3%), whereas falls from a height caused OWF in approximately half of the patients in the preschool pediatric group (42.9%). Concomitant injuries and facial fractures had a tendency to occur more frequently in the school-aged pediatric group. CONCLUSION: Significant differences according to the sex and mechanisms of injury were identified in preschool and school-aged pediatric patients with OWF.

Patterns and injuries associated with orbital wall fractures in elderly patients who visited the emergency room: a retrospective case-control study.

OBJECTIVES: This study aimed to determine orbital wall fracture (OWF) patterns and associated facial injuries in elderly patients and compare them with those in their younger adult counterparts. DESIGN: A retrospective case-control study. SETTING: An emergency department of a university-affiliated hospital located in an urban area. PARTICIPANTS: A total of 1378 adult patients with OWF diagnosed by CT from 1 January 2004 through 31 March 2014 were enrolled. Patients were categorised into elderly (≥65 years) and non-elderly (<65 years) groups. RESULTS: The elderly group (n=146) had a mean age of 74.0 years compared with 37.5 years in the non-elderly group (n=1232). Slipping was the most common cause of OWF in the elderly group (43.8%, p<0.001), whereas violence was the most common cause in the non-elderly group (37.3%, p<0.001). The lateral orbital wall was the more common site of fracture in the elderly group, and their injuries were more often associated with concurrent facial bone fractures, including the mandible, maxilla and zygoma, compared with the non-elderly group. After adjusting for sex and the mechanism of injury, inclusion in the elderly group was a significant risk factor for fracture of the lateral wall (OR 1.658; 95% CI 1.074 to 2.560) and concomitant facial bone fractures of the maxilla (OR 1.625; 95% CI 1.111 to 2.377) and zygoma (OR 1.670; 95% CI 1.126 to 2.475). CONCLUSIONS: Elderly patients were vulnerable to facial trauma, and concurrent facial bone fracture associated with OWF was more commonly observed in this age group. Therefore, a high index of suspicion and thorough investigation, including CT, for OWF-associated facial bone fractures are important.

Two helices in the third intracellular loop determine anoctamin 1 (TMEM16A) activation by calcium.

Anoctamin 1 (ANO1)/TMEM16A is a Cl(-) channel activated by intracellular Ca(2+) mediating numerous physiological functions. However, little is known of the ANO1 activation mechanism by Ca(2+). Here, we demonstrate that two helices, "reference" and "Ca(2+) sensor" helices in the third intracellular loop face each other with opposite charges. The two helices interact directly in a Ca(2+)-dependent manner. Positively and negatively charged residues in the two helices are essential for Ca(2+)-dependent activation because neutralization of these charges change the Ca(2+) sensitivity. We now predict that the Ca(2+) sensor helix attaches to the reference helix in the resting state, and as intracellular Ca(2+) rises, Ca(2+) acts on the sensor helix, which repels it from the reference helix. This Ca(2+)-dependent push-pull conformational change would be a key electromechanical movement for gating the ANO1 channel. Because chemical activation of ANO1 is viewed as an alternative means of rescuing cystic fibrosis, understanding its gating mechanism would be useful in developing novel treatments for cystic fibrosis.

TRPM2 mediates the lysophosphatidic acid-induced neurite retraction in the developing brain.

Intracellular Ca(2+) signal is a key regulator of axonal growth during brain development. As transient receptor potential (TRP) channels are permeable to Ca(2+) and mediate numerous brain functions, it is conceivable that many TRP channels would regulate neuronal differentiation. We therefore screened TRP channels that are involved in the regulation of neurite growth. Among the TRP channels, the Trpm2 level was inversely associated with neurite growth. TRPM2 was highly expressed in embryonic brain. Pharmacological perturbation or knockdown of TRPM2 markedly increased the axonal growth, whereas its overexpression inhibited the axonal growth. Addition of ADP ribose, an endogenous activator of TRPM2, to PC12 cells significantly repressed the axonal growth. TRPM2 was actively involved in the neuronal retraction induced by cerebrospinal fluid-rich lysophosphatidic acid (LPA). More importantly, neurons isolated from the brain of Trpm2-deficient mice have significantly longer neurites with a greater number of spines than those obtained from the brain of wild-type mice. Therefore, we conclude that TRPM2 mediates the LPA-induced suppression of axonal growth, which provides a long-sought mechanism underlying the effect of LPA on neuronal development.

Anoctamin 1 contributes to inflammatory and nerve-injury induced hypersensitivity.

BACKGROUND: Various pathological conditions such as inflammation or injury can evoke pain hypersensitivity. That represents the response to innocuous stimuli or exaggerated response to noxious stimuli. The molecular mechanism based on the pain hypersensitivity is associated with changes in many of ion channels in dorsal-root ganglion (DRG) neurons. Anoctamin 1 (ANO1/TMEM16A), a Ca2+ activated chloride channel is highly visible in small DRG neurons and responds to heat. Mice with an abolished function of ANO1 in DRG neurons demonstrated attenuated pain-like behaviors when exposed to noxious heat, suggesting a role in acute thermal nociception. In this study, we further examined the function of ANO1 in mediating inflammation- or injury-induced hyperalgesia or allodynia. RESULTS: Using Advillin/Ano1fl/fl (Adv/Ano1fl/fl) mice that have a functional ablation of Ano1 mainly in DRG neurons, we were able to determine its role in mediating thermal hyperalgesia and mechanical allodynia induced by inflammation or nerve injury. The thermal hyperalgesia and mechanical allodynia induced by carrageenan injection and spared-nerve injury were significantly reduced in Adv/Ano1fl/fl mice. In addition, flinching or licking behavior after bradykinin or formalin injection was also significantly reduced in Adv/Ano1fl/fl mice. Since pathological conditions augment nociceptive behaviors, we expected ANO1's contribution to the excitability of DRG neurons. Indeed, the application of inflammatory mediators reduced the threshold for action potential (rheobase) or time for induction of the first action potential in DRG neurons isolated from control (Ano1fl/fl) mice. These parameters for neuronal excitability induced by inflammatory mediators were not changed in Adv/Ano1fl/fl mice, suggesting an active contribution of ANO1 in augmenting the neuronal excitability. CONCLUSIONS: In addition to ANO1's role in mediating acute thermal pain as a heat sensor, ANO1 is also capable of augmenting the excitability of DRG neurons under inflammatory or neuropathic conditions and thereby aggravates inflammation- or tissue injury-induced pathological pain.

Axonal neuropathy-associated TRPV4 regulates neurotrophic factor-derived axonal growth.

Spinal muscular atrophy and hereditary motor and sensory neuropathies are characterized by muscle weakness and atrophy caused by the degenerations of peripheral motor and sensory nerves. Recent advances in genetics have resulted in the identification of missense mutations in TRPV4 in patients with these hereditary neuropathies. Neurodegeneration caused by Ca(2+) overload due to the gain-of-function mutation of TRPV4 was suggested as the molecular mechanism for the neuropathies. Despite the importance of TRPV4 mutations in causing neuropathies, the precise role of TRPV4 in the sensory/motor neurons is unknown. Here, we report that TRPV4 mediates neurotrophic factor-derived neuritogenesis in developing peripheral neurons. TRPV4 was found to be highly expressed in sensory and spinal motor neurons in early development as well as in the adult, and the overexpression or chemical activation of TRPV4 was found to promote neuritogenesis in sensory neurons as well as PC12 cells, whereas its knockdown and pharmacologic inhibition had the opposite effect. More importantly, nerve growth factor or cAMP treatment up-regulated the expression of phospholipase A(2) and TRPV4. Neurotrophic factor-derived neuritogenesis appears to be regulated by the phospholipase A(2)-mediated TRPV4 pathway. These findings show that TRPV4 mediates neurotrophic factor-induced neuritogenesis in developing peripheral nerves. Because neurotrophic factors are essential for the maintenance of peripheral nerves, these findings suggest that aberrant TRPV4 activity may lead to some types of pathology of sensory and motor nerves.

Slug, mammalian homologue gene of Drosophila escargot, promotes neuronal-differentiation through suppression of HEB/daughterless.

At the neuron developmental stage, neuron-precursor cells can be differentiated into neuron or glia cells. However, precise molecular mechanism to determine the cell fate has not been clearly demonstrated. In this study, we reveal that Drosophila esgarcot and its mammalian homologue genes, Snail and Slug, play a key role in neuronal differentiation. In Drosophila model system, overexpression of Esg, like as Wingless, suppresses the bristle formation. In contrast, elimination of Esg though RNAi promotes double bristle phenotype. We can also observe the similar phenotype in Snail-overexpression system. In mammalian system, overexpression of Slug or Snail can induce neuronal differentiation. Esg and its mammalian homologue gene Slug directly interact with Daughtherless and its mammalian homologue HEB and eliminate them through siah-1 mediated protein degradation. Thus, overexpression of siah-1 can promote neuron cell differentiation, whereas si-siah-1 blocks the Slug-induced HEB suppression. In fact, Drosophila SINA, Siah-1 homologue, has been also known to be involved in bristle formation and Neuronal differentiation. In addition, it has been revealed that CK1 is involved in Esg or Snail stability and Neuronal differentiation. However, Snail is regulated only by CK1 but not by Siah. Considering the fact that Slug mutations have been found in human genetic disease, waardenberg syndrome, major symptoms of which is loss of hearing neuron and odd eye, our result implies that slug/Snail system is required for proper neuronal differentiation, like as Esg in Drosophila.

[A case of renal vein thrombosis in a patient with ulcerative colitis].

Venous thrombosis and thromboembolism appear to occur more often in patient with inflammatory bowel disease (IBD). The cause of thrombotic complications in IBD is generally considered to be associated with hypercoagulable conditions. Its prevalence rate ranges from 1% to 8% in clinical studies and rises to 39% in autopsy, but the renal vein thrombosis is very rare complication in ulcerative colitis patient. A 24-year-old man presented with intermittent abdominal pain and hematochezia for 6 months and recently developed pitting edema for few weeks. He was diagnosed as severe ulcerative colitis involving whole colon combined with thrombosis in both renal veins by colonoscopy and computed tomography scan of abdomen. We used steroid for the treatment of ulcerative colitis and both intravenous lower molecular weight heparin and warfarin for renal vein thrombosis. His symptoms were improved after treatment and maintained with mesalazine and warfarin. Follow-up abdominal CT scan showed complete resolution of both renal vein thrombosis. Currently he has been followed up for 2 years with oral mesalazine.

Big brain, a Drosophila homologue of mammalian aquaporin, is regulated by the DRE/DREF system.

Drosophila big brain (bib) encodes for a protein similar to members of the major intrinsic protein family, which includes the water- and ion-conducting aquaporin (AQP) channels. In mammals, AQP dysregulation has been implicated in a variety of diseases, including colorectal cancer and colonic injury. However, the regulatory mechanisms of AQP expression remain to be clearly elucidated. In this study, as we found a DREF binding site (DRE) in the 5'-flanking regions of both the Drosophila bib gene and the human AQP1 gene, we assessed the role of DREF in bib gene expression. DREF in Drosophila and humans has been demonstrated to function as a key transcriptional activator for cell proliferation-related genes. Herein, we demonstrate that the DRE is required for optimal promoter activity of Drosophila bib gene, particularly in the larval imaginal discs, which are actively proliferating tissues, as well as the adult hindgut. Our results may provide insight into the mechanisms inherent to the regulation of mammalian AQP genes.

Pulsed-UV light inactivation of Cryptosporidium parvum.

Cryptosporidium parvum is an organism that threatens public health in the water industry. It is critical to develop improved detection methods as well as disinfection methods for protecting against cryptosporidiosis, which is caused by C. parvum. In this study, we investigated the ability of pulsed-light irradiation at 200-900 nm to inactivate C. parvum. Absolute quantitative real-time PCR was performed with cDNA made from total RNA extracted from C. parvum oocysts or HCT-8 cells infected with C. parvum oocysts in vitro. C. parvum oocysts in 100-mL quartz flasks were positioned 20, 30, and 40 cm from the light source, and the duration of irradiation was either 5 or 60 s. The reductions in oocyst viability (4.9 log10) and infectivity (6 log10) were maximal when the C. parvum oocysts were irradiated 20 cm from the pulsed-light source for 60 s, for which the UV dose was 278 mJ/cm2. The minimum dose of pulsed-UV light required for effective reduction in C. parvum infectivity (2 log10) was 15 mJ/cm2, which was achieved by 5 s of irradiation at 30 cm from the light source. This study confirmed that short-duration pulsed-UV light is an effective disinfection measure for C. parvum.

Age-related changes in Drosophila midgut are associated with PVF2, a PDGF/VEGF-like growth factor.

Age-associated changes in stem cell populations have been implicated in age-related diseases, including cancer. However, little is known about the underlying molecular mechanisms that link aging to the modulation of adult stem cell populations. Drosophila midgut is an excellent model system for the study of stem cell renewal and aging. Here we describe an age-related increase in the number and activity of intestinal stem cells (ISCs) and progenitor cells in Drosophila midgut. We determined that oxidative stress, induced by paraquat treatment or loss of catalase function, mimicked the changes associated with aging in the midgut. Furthermore, we discovered an age-related increase in the expression of PVF2, a Drosophila homologue of human PDGF/VEGF, which was associated with and required for the age-related changes in midgut ISCs and progenitor cell populations. Taken together, our findings suggest that PDGF/VEGF may play a central role in age-related changes in ISCs and progenitor cell populations, which may contribute to aging and the development of cancer stem cells.

Transcriptional regulation of the Drosophila ANT gene by the DRE/DREF system.

Adenine nucleotide translocase (ANT) is a crucial component in the maintenance of cellular energy homeostasis, as well as in the formation of the mitochondrial permeability transition pores. However, the molecular mechanisms regulating the expression of the ANT gene are poorly understood. In this study, we have identified three DNA replication-related elements (DRE; 5'-TATCGATA) in the 5'-flanking region of the Drosophila ANT (dANT) gene. Gel-mobility shift analyses revealed that all three of the DREs were recognized by the DRE-binding factor (DREF). The site-directed mutagenesis of these DRE sites induces a considerable reduction in the activity of the dANT gene promoter in vitro. Analyses with transgenic flies harboring a dANT-lacZ fusion gene bearing the wild-type or mutant DRE sites showed that the DRE sites were required for the expression of dANT in vivo. We determined that the over-expression or knockdown of DREF exerts a regulatory effect on the activity of the dANT promoter. In addition, we observed the collapse of mitochondrial membrane potential in the eye imaginal discs in which DREF was over-expressed. These results show that DRE/DREF is a crucial regulator of dANT gene expression, and also suggest the possibility that cross-talk may occur between the DRE/DREF system and mitochondrial functioning.