Retroductal Delivery of Epidermal Growth Factor Protects Salivary Progenitors after Irradiation.

Salivary gland hypofunction after irradiation is associated with a deficit of epithelial stem/progenitors in salivary glands. Although epidermal growth factor (EGF) is known to stimulate the proliferation of epithelial cells, the therapeutic effect of EGF on salivary epithelial stem/progenitors remains undetermined. In this study, we administered EGF to submandibular glands (SMGs) via a retrograde route through the SMG excretory duct before fractionated irradiation and examined whether EGF could protect salivary epithelial progenitor cells from radiation and alleviate radiation-induced salivary hypofunction. EGF-treated mice exhibited greater body and gland weights at 12 wk after irradiation than untreated mice. The retroductal delivery of EGF improved salivary secretory function and increased salivary amylase activity in a dose-dependent manner. Histological examinations highlighted the amelioration of the loss of keratine-14+ (KRT14+) basal ductal and/or MIST1+ acinar cells, as well as induction of fibrosis, following irradiation in EGF-treated mice. An additional in vitro experiment using a salivary gland organoid irradiation model indicated that the radioprotective effects of EGF promoted the growth and inhibited the apoptotic cell death of salivary epithelial cells. Our results suggest that retroductal delivery of EGF may be a promising therapeutic option for preventing radiation-induced salivary gland hypofunction.

Functional characterization of O-methyltransferases used to catalyse site-specific methylation in the post-tailoring steps of pradimicin biosynthesis.

AIMS: To identify the roles of the two O-methyltransferase homologous genes pdmF and pdmT in the pradimicin biosynthetic gene cluster of Actinomadura hibisca P157-2. METHODS AND RESULTS: Pradimicins are pentangular polyphenol antibiotics synthesized by bacterial type II polyketide synthases (PKSs) and tailoring enzymes. Pradimicins are naturally derivatized by combinatorial O-methylation at two positions (i.e., 7-OH and 11-OH) of the benzo[α]naphthacenequinone structure. PdmF and PdmT null mutants (PFKO and PTKO) were generated. PFKO produced the 11-O-demethyl shunt metabolites 11-O-demethylpradimicinone II (1), 11-O-demethyl-7-methoxypradimicinone II (2), 11-O-demethylpradimicinone I (3) and 11-O-demethylpradimicin A (4), while PTKO generated the 7-O-demethyl derivatives pradimicinone II (5) and 7-hydroxypradimicin A (6). Pradimicinones 1, 2, 3, and 5 were fed to a heterologous host Escherichia coli harbouring expression plasmid pET-22b::pdmF or pET-28a::pdmT. PdmF catalysed 11-O-methylation of pradimicinones 1, 2, and 3 regardless of O-methylation at the C-7 position, while PdmT was unable to catalyse 7-O-methylation when the C-11 hydroxyl group was methylated (5). CONCLUSIONS: PdmF and PdmT were involved in 11-O- and 7-O-methylations of the benzo[α]naphthacenequinone moiety of pradimicin, respectively. Methylation of the C-7 hydroxyl group precedes methylation of the C-11 hydroxyl group in pradimicin biosynthesis. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first reported demonstration of the functions of PdmF and PdmT for regiospecific O-methylation, which contributes to better understanding of the post-PKS modifications in pradimicin biosynthesis as well as to rational engineering of the pradimicin biosynthetic machinery.

Arctic sea ice melt leads to atmospheric new particle formation.

Atmospheric new particle formation (NPF) and growth significantly influences climate by supplying new seeds for cloud condensation and brightness. Currently, there is a lack of understanding of whether and how marine biota emissions affect aerosol-cloud-climate interactions in the Arctic. Here, the aerosol population was categorised via cluster analysis of aerosol size distributions taken at Mt Zeppelin (Svalbard) during a 11 year record. The daily temporal occurrence of NPF events likely caused by nucleation in the polar marine boundary layer was quantified annually as 18%, with a peak of 51% during summer months. Air mass trajectory analysis and atmospheric nitrogen and sulphur tracers link these frequent nucleation events to biogenic precursors released by open water and melting sea ice regions. The occurrence of such events across a full decade was anti-correlated with sea ice extent. New particles originating from open water and open pack ice increased the cloud condensation nuclei concentration background by at least ca. 20%, supporting a marine biosphere-climate link through sea ice melt and low altitude clouds that may have contributed to accelerate Arctic warming. Our results prompt a better representation of biogenic aerosol sources in Arctic climate models.

Association of lipid parameters and insulin resistance with bone health in South Korean adolescents.

UNLABELLED: This study investigated the association between lipid profiles and insulin resistance and bone mineral content (BMC) in Korean adolescents and found that BMC was inversely associated with triglyceride (TG) and homeostasis model assessment of insulin resistance (HOMA-IR). This association did not differ according to obesity status in either boys or girls. INTRODUCTION: To prevent future osteoporosis, it is important to identify factors that affect bone health in adolescents as well as adults. This study aimed to examine the association between lipid profiles and insulin resistance and BMC in Korean adolescents. METHODS: Data from 706 boys and 621 girls, who participated in the Korea National Health and Nutrition Examination Survey from 2008 to 2011, were analyzed. Lipid profiles were measured, and HOMA-IR was calculated to assess insulin resistance. BMC was measured for the total femur, femur neck, and lumbar spine by using whole-body dual-energy X-ray absorptiometry (DXA). RESULTS: TG level and HOMA-IR were negatively correlated with BMC at all three sites in boys. In girls, TG level showed a negative correlation with BMC at the femur neck and lumbar spine, and HOMA-IR was negatively associated with BMC at the femur neck only. These inverse associations did not differ according to obesity status in either sex. Adjusted means of BMC at the three sites in boys tended to decrease in the higher tertile groups of TG and HOMA-IR, and the adjusted means of BMC for the total femur in girls tended to decrease in the higher tertile groups of TG and HOMA-IR. CONCLUSIONS: BMC was inversely associated with TG and HOMA-IR in Korean adolescents, and this association was more pronounced in boys. This association did not differ according to obesity status in either sex.

Sudden deafness as an initial presenting symptom of posterior inferior cerebellar artery infarction: two case reports.

OBJECTIVE: This paper reports on two patients with posterior inferior cerebellar artery infarction whose only presenting complaint was acute unilateral hearing loss. CASE REPORTS: In the two cases reported, sudden hearing loss was an initial symptom, with no other neurological signs. Infarction in the territory of the posterior inferior cerebellar artery was diagnosed using brain magnetic resolution imaging. The patients had some degree of hearing improvement 3 or 4 days after initial treatment. CONCLUSION: In this article, new cases of posterior inferior cerebellar artery infarction presenting as sudden deafness, without prominent neurological signs, are described. Otologists should be aware that hearing loss can sometimes appear as a warning sign of impending posterior inferior cerebellar artery infarction.

Large-scale generation of cell-derived nanovesicles.

Exosomes are enclosed compartments that are released from cells and that can transport biological contents for the purpose of intercellular communications. Research into exosomes is hindered by their rarity. In this article, we introduce a device that uses centrifugal force and a filter with micro-sized pores to generate a large quantity of cell-derived nanovesicles. The device has a simple polycarbonate structure to hold the filter, and operates in a common centrifuge. Nanovesicles are similar in size and membrane structure to exosomes. Nanovesicles contain intracellular RNAs ranging from microRNA to mRNA, intracellular proteins, and plasma membrane proteins. The quantity of nanovesicles produced using the device is 250 times the quantity of naturally secreted exosomes. Also, the quantity of intracellular contents in nanovesicles is twice that in exosomes. Nanovesicles generated from murine embryonic stem cells can transfer RNAs to target cells. Therefore, this novel device and the nanovesicles that it generates are expected to be used in exosome-related research, and can be applied in various applications such as drug delivery and cell-based therapy.

Therapeutic targeting of epigenetic components in amyotrophic lateral sclerosis (ALS).

Amyotrophic lateral sclerosis (ALS) is an adult-onset motor neuron disease characterized by degeneration of motor neuron and glial activation followed by the progressive muscle loss and paralysis. Numerous distinct therapeutic interventions have been examined but currently ALS does not have a cure or an efficacious treatment for the disorder. Glutamate- induced excitotoxicity, inflammation, mitochondrial dysfunction, oxidative stress, protein aggregation, transcription deregulation, and epigenetic modifications are associated with the pathogenesis of ALS and known to be therapeutic targets in ALS. In this review, we discuss translational pharmacological studies targeting epigenetic components to ameliorate ALS. Understanding of the epigenetic mechanisms will provide novel insights that will further identify potential biological markers and therapeutic approaches for treating ALS. A combination of treatments that modulate epigenetic components and multiple targets may prove to be the most effective therapy for ALS.

Regio-selectively reduced streptogramin A analogue, 5,6-dihydrovirginiamycin M1 exhibits improved potency against MRSA.

A newly reduced macrocyclic lactone antibiotic streptogramin A, 5,6-dihydrovirginiamycin M1 was created by feeding virginiamycin M1 into a culture of recombinant Streptomyces venezuelae. Its chemical structure was spectroscopically elucidated, and this streptogramin A analogue showed twofold higher antibacterial activities against methicillin-resistant Staphylococcus aureus (MRSA) compared with its parent molecule virginiamycin M1. Docking studies using the model of streptogramin A acetyltransferase (VatA) suggested that the newly generated analogue binds tighter with overall lower free energy compared with the parent molecule virginiamycin M1. This hypothesis was validated experimentally through the improvement of efficacy of the new analogue against MRSA strains. The biotransformation approach presented herein could have a broad application in the production of reduced macrocyclic molecules.

Case of bilateral pneumolabyrinth presenting as sudden, bilateral deafness, without temporal bone fracture, after a fall.

OBJECTIVE: We report a case of bilateral pneumolabyrinth presenting as sudden, bilateral deafness, without temporal bone fracture, after a fall. CASE REPORT: A 49-year-old man presented with sudden, bilateral deafness and whirling vertigo, without any other neurological manifestations. Temporal bone computed tomography clearly demonstrated the presence of air in the vestibule and cochlea on both sides. However, there was no definite fracture line, ossicular chain anomaly or soft tissue density in the temporal bone or middle-ear cavity. The patient was treated conservatively. Unfortunately, there was no improvement in his hearing. CONCLUSION: Pneumolabyrinth is an uncommon condition in which air is present in the vestibule or cochlea. It is rarely found, even with fractures violating the otic capsule or with transverse fractures of the temporal bone. In addition, its bilateral occurrence is extremely rare. In this article, we describe a case of bilateral pneumolabyrinth presenting as sudden, bilateral deafness, without temporal bone fracture, an occurrence which has not previously been reported.

Spatial distribution of antimicrobial peptides and mast cells in the skin of the external auditory canal.

OBJECTIVE: The direct activity of antimicrobial peptides against microbes is thought to be an essential first line of defence in the skin; however, little is known about antimicrobial peptide secretion in the skin of the external auditory canal. Evidence suggests that mast cells contribute to the secretion of antimicrobial peptides. This study aimed to examine the distribution of mast cells and antimicrobial peptides, including human ß-defensin-1 and -2 and LL-37, in the external auditory canal skin. METHODS: External auditory canal skin samples from 12 patients undergoing middle-ear surgery with canaloplasty were immunohistochemically stained to detect expression of mast cell markers (tryptase and chymase) and antimicrobial peptides (human ß-defensin-1 and -2 and LL-37). RESULTS: Mast cells and human ß-defensin-1 were present in the ceruminous glands but not in the sebaceous glands. The increased presence of mast cells, human ß-defensin-1 and LL-37 in ceruminous glands suggests that mast cells may participate in the secretion of antimicrobial peptides from ceruminous glands. CONCLUSION: These findings suggest that mast cells contribute to the secretion of antimicrobial peptides in the ceruminous glands of the external auditory canal skin.

Effectiveness of the Streamlined Liner of the Pharynx Airway (SLIPATM) in allowing positive pressure ventilation during gynaecological laparoscopic surgery.

The aim of this study was to assess whether the Streamlined Liner of the Pharynx Airway (SLIPA) performed as well as an endotracheal tube for positive pressure ventilation in gynaecological laparoscopic surgery in the Trendelenburg position. Forty patients (American Society of Anesthesiologists physical status I to III) were randomly divided into two groups: SLIPA (n = 20) or endotracheal tube group (n = 20). Lung mechanics and severity of postoperative sore throat were assessed in both groups. In the SLIPA group, the oropharyngeal leak pressure was also measured. There were no significant differences between groups in the lung mechanics. In the SLIPA group, oropharyngeal leak pressure and peak inspiratory pressure increased significantly after gas insufflation compared to 10 minutes after patient positioning in the lithotomy position (P < 0.05). The difference between oropharyngeal leak pressure and peak inspiratory pressure (approximately 10 cmH2O throughout the procedure) remained suitable for airway maintenance. The incidence of sore throat was similar in both groups but the severity was less in the SLIPA group 24 hours after surgery (P < 0.05). There were no other complications such as regurgitation noted in either group. In the study population, the SLIPA performed as well as an endotracheal tube in allowing positive pressure ventilation without gas leak during gynaecological laparoscopy. The way in which the SLIPA increases its resistance to gas leak as the inspiratory pressure rises may account for this.

Methylenetetrahydrofolate reductase C677T gene mutation as risk factor for sudden sensorineural hearing loss: association with plasma homocysteine, folate and cholesterol concentrations.

OBJECTIVE: Impaired cochlear perfusion appears to be the most important event in the development of sudden sensorineural hearing loss. Methylenetetrahydrofolate reductase gene mutations at nucleotide 677 cause reduced methylenetetrahydrofolate reductase enzyme activity, resulting in vascular impairment. METHODS: Thirty-three patients and 68 control subjects underwent audiological and haematological investigation. RESULTS: No statistically significant association was found between sudden sensorineural hearing loss and the methylenetetrahydrofolate reductase C677T gene mutation. Mean homocysteine and cholesterol concentrations were significantly higher in patients than in controls. Mean folate levels were significantly lower in patients than in controls. Amongst patients with sudden sensorineural hearing loss, no significant differences in mean cholesterol, homocysteine or folate concentration were found, comparing patients with methylenetetrahydrofolate reductase C677T mutation genotypes with those without. CONCLUSION: No statistically significant association was found between the methylenetetrahydrofolate reductase C677T gene mutation and sudden sensorineural hearing loss. There was a statistically significant difference between the homocysteine, folate and cholesterol concentrations of sudden sensorineural hearing loss patients, compared with controls. However, there was no statistically significant difference in these levels, comparing patients with and without the methylenetetrahydrofolate reductase C677T mutation.

Performance of electron beam irradiation for treatment of groundwater contaminated with acetone.

The purposes of this study were to evaluate the efficiency of acetone removal by electron beam irradiation in groundwater and the effect of various conditions. According to the results, the removal kinetics of acetone were pseudo first-order, and the removal efficiencies were expressed to the (%) removal and G-values. By adding sulfite, it was confirmed that acetone was mainly degraded by the reaction with the hydrated electrons. The presence of nitrate caused the removal of acetone to decrease. But there was no significant effect of alkalinity on the removal of acetone. The effect of the initial pH values (pH 5 to 9) on the acetone removal efficiency was negligible, but the pH value decreases due to the formation of acidic compounds after irradiation. Consequently, the radiation-induced removal reactions of acetone followed the pseudo-first-order kinetic model; in addition to the initial concentration of acetone, nitrate and the absorbed dose were important factors in removing acetone from an aqueous solution using electron beam irradiation. The effects of general pH and alkalinity on the degrading acetone were negligible.

In vitro metabolism of a novel PPAR gamma agonist, KR-62980, and its stereoisomer, KR-63198, in human liver microsomes and by recombinant cytochrome P450s.

1. KR-62980 and its stereoisomer KR-63198 are novel and selective peroxisome proliferator-activated receptor gamma (PPAR gamma) modulators with activity profiles different from that of rosiglitazone. This study was performed to identify the major metabolic pathways for KR-62980 and KR-63198 in human liver microsomes. 2. Human liver microsomal incubation of KR-62980 and KR-63198 in the presence of a beta-nicotinamide adenine dinucleotide phosphate (NADPH)-generating system resulted in hydroxy metabolite formation. In addition, the specific cytochrome P450s (CYPs) responsible for KR-62980 and KR-63198 hydroxylation were identified by using a combination of chemical inhibition in human liver microsomes and metabolism by recombinant P450s. It is shown that CYP1A2, CYP2D6, CYP3A4, and CYP3A5 are the predominant enzymes in the hydroxylation of KR-62980 and KR-63198. 3. The intrinsic clearance through hydroxylation was consistently and significantly higher for KR-62980 than for KR-63198, indicating metabolic stereoselectivity (CL(int) of 0.012 +/- 0.001 versus 0.004 +/- 0.001 microl min(-1) pmol(-1) P450, respectively). 4. In a drug-drug interaction study, KR-62980 and KR-63198 had no effect on the activities of the P450s tested (IC(50) > 50 microM), suggesting that in clinical interactions between KR-62980 and KR-63198 the P450s tested would not be expected.

Sequence-based screening for self-sufficient P450 monooxygenase from a metagenome library.

AIMS: Cytochrome P450 monooxygenases (CYPs) are useful catalysts for oxidation reactions. Self-sufficient CYPs harbour a reductive domain covalently connected to a P450 domain and are known for their robust catalytic activity with great potential as biocatalysts. In an effort to expand genetic sources of self-sufficient CYPs, we devised a sequence-based screening system to identify them in a soil metagenome. METHODS AND RESULTS: We constructed a soil metagenome library and performed sequence-based screening for self-sufficient CYP genes. A new CYP gene, syk181, was identified from the metagenome library. Phylogenetic analysis revealed that SYK181 formed a distinct phylogenic line with 46% amino-acid-sequence identity to CYP102A1 which has been extensively studied as a fatty acid hydroxylase. The heterologously expressed SYK181 showed significant hydroxylase activity towards naphthalene and phenanthrene as well as towards fatty acids. CONCLUSIONS: Sequence-based screening of metagenome libraries is expected to be a useful approach for searching self-sufficient CYP genes. The translated product of syk181 shows self-sufficient hydroxylase activity towards fatty acids and aromatic compounds. SIGNIFICANCE AND IMPACT OF THE STUDY: SYK181 is the first self-sufficient CYP obtained directly from a metagenome library. The genetic and biochemical information on SYK181 are expected to be helpful for engineering self-sufficient CYPs with broader catalytic activities towards various substrates, which would be useful for bioconversion of natural products and biodegradation of organic chemicals.

A novel scratching approach for measuring age-related changes in the in situ toughness of bone.

A scratch test using a nanoindentation system was proposed in this study to assess the age-related changes in the in situ toughness of bone matrix at ultrastructural levels. A tissue removal energy density (u(r)) was defined and estimated as the work done by the scratch (U(T)) divided by the total volume of the scratch groove (u(s)). The value of u(s) was used as a relative measure of the in situ toughness of the tissue. Human cortical bone specimens obtained from middle-aged (between 49 and 59 years old) and elderly groups (over 69 years old) were tested using this technique. A significant difference in the estimated removal energy density (u(s)) in the secondary osteons was found between the middle-aged and elderly groups (5.49+/-0.696 vs. 4.09+/-1.30 N/mm(2), respectively).

RNA polymerase beta-subunit gene (rpoB) sequence analysis for the identification of Bacteroides spp.

Partial rpoB sequences (317 bp) of 11 species of Bacteroides, two Porphyromonas spp. and two Prevotella spp. were compared to delineate the genetic relationships among Bacteroides and closely related anaerobic species. The high level of inter-species sequence dissimilarities (7.6-20.8%) allowed the various Bacteroides spp. to be distinguished. The position of the Bacteroides distasonis and Bacteriodes merdae cluster in the rpoB tree was different from the position in the 16S rRNA gene tree. Based on rpoB sequence similarity and clustering in the rpoB tree, it was possible to correctly re-identify 80 clinical isolates of Bacteroides. In addition to two subgroups, cfiA-negative (division I) and cfiA-positive (division II), of Bacteroides fragilis isolates, two distinct subgroups were also found among Bacteroides ovatus and Bacteroides thetaiotaomicron isolates. Bacteroides genus-specific rpoB PCR and B. fragilis species-specific rpoB PCR allowed Bacteroides spp. to be differentiated from Porphyromonas and Prevotella spp., and also allowed B. fragilis to be differentiated from other non-fragilisBacteroides spp. included in the present study.

Avermectin: biochemical and molecular basis of its biosynthesis and regulation.

Avermectin and its analogues, produced by Streptomyces avermitilis, are major commercial antiparasitic agents in the field of animal health, agriculture, and human infections. They are 16-membered pentacyclic lactone compounds derived from polyketide and linked to a disaccharide of the methylated deoxysugar l-oleandrose. Labeling studies, analyses of the biosynthetically blocked mutants, and the identification of the avermectin gene cluster allows characterization of most of the biosynthetic pathway. Recent completion of S. avermitilis genome sequencing is also expected to help in revealing the precise biosynthetic sequence and the complicated regulatory mechanism for avermectin biosynthesis, which has been long-awaited to be elucidated. The well characterized avermectin biosynthetic pathway and availability of S. avermitilis genome information in combination with the recent development of combinatorial biosynthesis should allow us to redesign more potent avermectin analogues and to engineer S. avermitilis as a more efficient host for the production of important commercial analogues.

Estimation of the effective transversely isotropic elastic constants of a material from known values of the material's orthotropic elastic constants.

A method is illustrated for determining the effective transversely isotropic (or isotropic) elastic constants from measured orthotropic elastic constants. This method consists of constructing upper and lower bounds on the effective transversely isotropic (or isotropic) elastic constants using the known orthotropic values. This method is illustrated using three sets of elastic constants for bone. Fortunately, the upper and lower bounds are very close. Thus very good approximations for the effective transversely isotropic (or isotropic) elastic constants for cortical and cancellous bone are obtained from previously published data on the orthotropic elastic constants for those tissue types. This work is undertaken to build a greater database for the transversely isotropic elastic constants of bone with the intention of employing them in a transversely isotropic model of bone poroelasticity. An interesting aspect of the present result is that the Voigt and Reuss bounds are very tight for these anisotropic materials. This is not always the case for these bounds.

Effect of head rotation on lateral cephalometric radiographs.

The purpose of this study was to identify the potential projection errors of lateral cephalometric radiographs due to head rotation in the vertical Z-axis. For this investigation, 17 human dry skull samples with permanent dentition were collected from the Department of Anatomy in the College of Medicine, Chosun University. They had no gross asymmetry and were well preserved. Each dry skull was rotated from 0 degrees to +/- 15 degrees at 1 degrees intervals. A vertical axis, the Z-axis, was used as a rotational axis to have 527 lateral cephalometric radiographs exposed. The findings were that: (1) angular measurements have fewer projection errors than linear measurements; (2) the greater the number of landmarks on the midsagittal plane that are included in angular measurements, the fewer the projection errors occurring; (3) horizontal linear measurements decrease gradually in length as the rotational angle toward the film increases, whereas a small increase and then decrease of the length occurs as the rotational angle toward the focal spot increases; (4) horizontal linear measurements have more projection errors than vertical linear measurements according to head rotation; and (5) projection errors of vertical linear measurements increase as the distance from the rotational axis increases. In summary, angular measurements of lateral cephalometric radiographs are more useful than linear measurements in minimizing the projection errors associated with head rotation on a vertical axis.