Fusarium euwallaceae sp. nov.--a symbiotic fungus of Euwallacea sp., an invasive ambrosia beetle in Israel and California.

The invasive Asian ambrosia beetle Euwallacea sp. (Coleoptera, Scolytinae, Xyleborini) and a novel Fusarium sp. that it farms in its galleries as a source of nutrition causes serious damage to more than 20 species of live trees and pose a serious threat to avocado production (Persea americana) in Israel and California. Adult female beetles are equipped with mandibular mycangia in which its fungal symbiont is transported within and from the natal galleries. Damage caused to the xylem is associated with disease symptoms that include sugar or gum exudates, dieback, wilt and ultimately host tree mortality. In 2012 the beetle was recorded on more than 200 and 20 different urban landscape species in southern California and Israel respectively. Euwallacea sp. and its symbiont are closely related to the tea shot-hole borer (E. fornicatus) and its obligate symbiont, F. ambrosium occurring in Sri Lanka and India. To distinguish these beetles, hereafter the unnamed xyleborine in Israel and California will be referred to as Euwallacea sp. IS/CA. Both fusaria exhibit distinctive ecologies and produce clavate macroconidia, which we think might represent an adaption to the species-specific beetle partner. Both fusaria comprise a genealogically exclusive lineage within Clade 3 of the Fusarium solani species complex (FSSC) that can be differentiated with arbitrarily primed PCR. Currently these fusaria can be distinguished only phenotypically by the abundant production of blue to brownish macroconidia in the symbiont of Euwallacea sp. IS/CA and their rarity or absence in F. ambrosium. We speculate that obligate symbiosis of Euwallacea and Fusarium, might have driven ecological speciation in these mutualists. Thus, the purpose of this paper is to describe and illustrate the novel, economically destructive avocado pathogen as Fusarium euwallaceae sp. nov. S. Freeman et al.

Premedication with cyclooxygenase-2 inhibitor meloxicam reduced postoperative pain in patients after oral surgery.

The efficacy of the selective cyclooxygenase-2 (COX-2) inhibitor meloxicam for treatment of postoperative oral surgical pain was assessed in a randomized controlled trial. Patients undergoing unilateral mandibular 3rd molar extraction surgery were allocated to 3 groups, A, B and C. After oral premedication of meloxicam 10 mg in group A, ampiroxicam 27 mg in group B and placebo in group C, surgery was completed within 30 min under local anaesthesia using 2% lidocaine. For postoperative pain relief the patients were allowed to take oral loxoprofen (60 mg per tablet). Postoperative pain was evaluated at the clinic on the 1st, 7th and 14th postoperative day (POD) using a visual analogue scale (VAS), as was the number of loxoprofen tablets consumed, and the results were compared among the 3 groups with statistical significance of P<0.05. VAS scores on 1 POD were significantly lower in group A than in group C. Loxoprofen consumption on the day of surgery and 1 POD was significantly lower in group A than in group C (P<0.01). Total analgesic consumption was significantly lower in groups A and B than in group C (P<0.02). The COX-2 inhibitor, meloxicam 10 mg used for premedication reduced postoperative pain compared with control in oral surgery.

An evaluation of polyvinylidene film dressing for treatment of pressure ulcers in older people.

OBJECTIVE: To compare the effectiveness of polyvinylidene (PVL) food wrap as a dressing material versus conventional ointments and gauze dressings for pressure ulcers in patients aged 60 years or over. METHODS: A prospective open controlled trial was conducted. The severity of pressure ulcers in both groups was evaluated using the DESIGN system. Scores were compared at baseline, four, eight and 12 weeks. RESULTS: Twenty-five patients were treated with the PVL film (test) dressings and 24 with the conventional treatment (control). The test group showed greater improvement in DESIGN scores than the control group throughout the observation period, and the difference reached statistical significance at 12 weeks (p < 0.05). The incidences of local wound infection in the two groups were not statistically different (p > 0.999). CONCLUSION: The PVL film dressing was more effective than conventional treatment in the management of severe pressure ulcers.

Influence of Nonenzymatic Glycation in Dentinal Collagen on Dental Caries.

Advanced glycation end-products (AGEs) are generated via nonenzymatic glycation of dentinal collagen, resulting in accumulation of AGEs in dentin tissue. Since accumulated AGEs cause crosslinking between amino acid polypeptides in the collagen molecule and modify mechanical properties of dentinal collagen, the authors assumed that there would be a significant interaction between the generation of AGEs and progression of caries in dentin. To confirm such an interaction, spectroscopic imaging analyses (i.e., nanosecond fluorescence lifetime imaging and second harmonic generation light imaging) were performed in addition to biochemical and electron microscopic analyses in the present study. Seven carious human teeth were fixed in paraformaldehyde and cut longitudinally into 1-mm sections using a low-speed diamond saw for the following analyses. In transmission electron microscopy (TEM) analysis, nondecalcified specimens were embedded in epoxy resin and sliced into thin sections for observation. For the immunohistochemical analysis, the specimens were paraffin embedded after decalcification for 2 wk and sectioned with a microtome. Resultant sections were stained with anti-AGE and anticollagen antibodies. The demineralized specimens were used for spectroscopic analyses without additional treatment. For Western blotting analysis, specimens were separated into carious and sound dentin. Each specimen was homogenized with a bead crusher and an ultrasonic homogenizer and then treated with hydrochloric acid. In carious dentin, the collagen fibers showed an amorphous structure in the TEM image, and the AGEs were localized in the areas of bacterial invasion in the immunostaining image. The total amount of AGEs in carious dentin was higher than in sound dentin in Western blotting. The ultrastructure of type I collagen and total amount of AGEs varied markedly in the dentinal caries region. The fluorescence lifetime was shorter in the carious area than that in the sound areas, indicating an increase of AGEs in the carious area. The increase of AGEs could influence the progression of dentinal caries.

Synthesis of antibiotic-loaded interporous hydroxyapatite blocks by vacuum method and in vitro drug release testing.

Interporous hydroxyapatite ceramic (Ca10(PO4)6(OH)2) has excellent bio-compatibility and interlinked pore structure, antibiotics could be loaded into pores in vacuum system. To confirm penetration of the agent to the HAb (2 cm3 cubic block), the aminoglycoside antibiotic (Isepamicin Sulfate; ISP) dissolved in eosin dye at various vacuum pressures. In ISP slow release study, the blocks were placed in 5 ml of PBS at a temperature of 37 degrees C. The PBS was replaced every 48 h and samples containing released ISP were stored until assay. All were found to release the drug maintaining a mean concentration of 0.41 microg ml(-1) even after 18 days of nine exchanges. This concentration of antibiotic exceeded the minimum inhibitory concentration against the common causative organisms of osteomyelitis. The results suggest that HAb impregnated with antibiotics using a simple vacuum system may serve as a valuable new method of administering local chemotherapy, primarily when used as a strut graft for bone defects.

Secretory function of adrenal chromaffin cells cultured on polypyrrole films.

Polypyrrole (PPy) is a conducting polymer and is obtained electrochemically on an electrode such as indium-tin oxide (ITO). In this study, in order to develop a novel cell-culture system which makes it possible to communicate with cultured mammalian cells, bovine adrenal chromaffin cells were cultured on PPy-coated ITO plates for 7 days and the influence of PPy-coating on the cell functions was investigated. Since the chromaffin cells synthesize and secrete catecholamines such as adrenaline and noradrenaline, the amount of synthesized and released catecholamines from the chromaffin cells cultured on PPy-coating and ITO itself were measured. The cells on the PPy-coated ITO plate could be kept in culture, without any significant changes in morphology and in the secretory responsiveness to acetylcholine as compared with those of the cells cultured on collagen. On the contrary, the cells on the ITO plate lost the responsiveness, while the amount of catecholamines synthesized was affected little by both PPy and ITO surfaces. It is suggested that PPy supports the secretory function of the chromaffin cells when they are cultured on it. This paper describes that PPy films are applicable as a polymer-modified electrode which support the cell function without collagen.

Change in cytoplasmic free Ca2+ concentration in rabbit platelets contacting with albumin coated and uncoated polystyrene surfaces.

Direct measurement of cytoplasmic free Ca2+ concentration in rabbit platelets in contact with polystyrene latexes was examined by monitoring an intracellular trapped fluorescent indicator dye, Fura 2, as a quantitative evaluation of platelet activation on material surfaces. It was found that an increase in cytoplasmic free calcium levels in platelets in contacting latexes was strongly reduced by the adsorption of albumin on the latex surfaces. Further, such an elevation of cytoplasmic free calcium levels was found to require a calcium influx from an extracellular medium across the platelet-plasma membrane.

In vitro and in vivo studies on antithrombogenicity of poly(propylene oxide) segmented nylon 610 in relation to its crystalline-amorphous microstructure.

In vitro and in vivo antithrombogenicity of poly(propylene oxide) segmented nylon 610 was investigated. For the in vitro simulation of thrombosis on copolymer surfaces, a modified microsphere column method evaluated the thrombosis time of the copolymer column. For the in vivo evaluation of the antithrombogenicity of copolymer surfaces, small diameter tubing, precoated with copolymer, was implanted in rabbits as an arteriovenous shunt to determine the time of patency for the copolymer tubing. In both the in vitro and in vivo tests, the copolymer having a particular size and distribution of crystalline and amorphous microphases on its surface exhibited excellent antithrombogenicity; the copolymer with a long period of approximately 12 nm and a crystallite thickness of 6.5 nm suppressed the activation process of adhering platelets to show thromboresistant property. This paper emphasizes the importance of controlling the crystalline-amorphous microstructure for the antithrombogenicity of polymer surfaces with a semicrystalline microstructure.

Wear resistance of experimental Ti-Cu alloys.

After using cast titanium prostheses in clinical dental practice, severe wear of titanium teeth has been observed. This in vitro study evaluated the wear behavior of teeth made with several cast titanium alloys containing copper (CP Ti+3.0 wt% Cu; CP Ti+5.0 wt% Cu; Ti-6Al-4V +1.0 wt% Cu; Ti-6Al-4V+4.0 wt% Cu) and compared the results with those for commercially pure (CP) titanium, Ti-6Al-4V, and gold alloy. Wear testing was performed by repeatedly grinding upper and lower teeth under flowing water in an experimental testing apparatus. Wear resistance was assessed as volume loss (mm(3)) at 5kgf (grinding force) after 50,000 strokes. Greater wear was found for the six types of titanium than for the gold alloy. The wear resistance of the experimental CP Ti+Cu and Ti-6Al-4V+Cu alloys was better than that of CP titanium and Ti-6Al-4V, respectively. Although the gold alloy had the best wear property, the 4% Cu in Ti-6Al-4V alloy exhibited the best results among the titanium metals. Alloying with copper, which introduced the alpha Ti/Ti(2)Cu eutectoid, seemed to improve the wear resistance.

Temperature-modulated platelet and lymphocyte interactions with poly(N-isopropylacrylamide)-grafted surfaces.

Temperature-responsive semitelechelic poly(N-isopropylacrylamide) (PIPAAm) bearing a carboxyl end group has been chemically immobilized on aminated polystyrene particle surfaces via condensation reaction. PIPAAm-grafted particles were uniformly suspended in aqueous media at lower temperatures. With increasing temperature, PIPAAm-grafted particles aggregated and precipitated. Such reversible changes in particle colloidal behaviour was correlated to temperature-modulated hydrophilic/hydrophobic changes of particle surfaces modified by PIPAAm hydration/dehydration with temperature changes. Interactions between platelets and PIPAAm-grafted surfaces were studied by monitoring cytoplasmic free Ca2+ concentration ([Ca2+]i) changes in platelets using intracellularly-trapped Ca2+ indicator dye, Fura 2, at various temperatures. Although changes in [Ca2+]i in platelets in contact with PIPAAm-grafted particles were not observed below the critical temperature of PIPAAm, significant changes in [Ca2+]i in platelets were induced by contact with particles above this critical temperature. Furthermore, temperature-modulated cell adsorption/desorption control by PIPAAm-grafted particles was investigated using a particle aggregation assay in the presence of lymphocytes. Below the critical temperature of PIPAAm, mixed suspensions were completely homogeneous due to minimal interaction between lymphocytes and hydrated particles. In contrast, aggregated precipitates were observed by increasing the suspension temperature above the critical temperature of PIPAAm resulting from strong hydrophobic interactions between particles with lymphocytes. These precipitates are reversibly resuspended in cold buffer. The feasibility of cell activation/inactivation or cell attachment/detachment control by temperature-modulated surface changes is attractive for suspension cell culture and drug delivery at targeted sites in vivo.

Temperature modulated solubility-activity alterations for poly(N-isopropylacrylamide)-lipase conjugates.

Chemical modification of proteins by use of functional polymers is expected to endow them with new properties without destroying their native functions, thus providing useful materials for application in different fields. We have synthesized poly(N-isopropylacrylamide) [poly(IPAAm)] co-oligomer with N,N-dimethylacrylamide (DMAAm) and reactive end groups by telomerization of IPAAm. This co-oligomer exhibits a lower critical solution temperature (LCST) at 37 degrees C. Using this temperature-responsive semitelechelic co-oligomer, we prepared polymer-enzyme conjugates of lipase by covalent coupling via carboxyl end-groups. This bioconjugate exhibits a LCST at 37 degrees C, having rapid, reversible hydration-dehydration changes due to highly mobile free polymer end groups. The conjugate retained its native enzymatic activity below this critical temperature, above which it precipitated and its catalytic function was shut off. This conjugate can be readily separated from reaction mixtures as a precipitate by simple temperature changes after reaction and reused in cycles without denaturation. Such a modulated system is attractive for application as a novel bioreactor system.

Immunohistochemical detection and distribution of enamelysin (MMP-20) in human odontogenic tumors.

Enamelysin is a tooth-specific protease that was initially isolated from porcine enamel organ and subsequently from human odontoblasts. Since this protease is thought to play important roles in tooth development, the evaluation of enamelysin in odontogenic tumors may aid our understanding of the histogenesis and cell differentiation of such lesions. A monoclonal antibody (203-1C7) was generated against synthesized human enamelysin oligopeptide and was used to assess the immunolocalization of enamelysin in healthy developing tooth germs and various types of odontogenic lesions. In tooth germs, enamelysin expression was detected only in the secretory enamel. Thus, 203-1C7 may serve as an enamel-specific marker in the late stage of enamel matrix development and calcification. In odontogenic lesions, strong enamelysin staining was demonstrated in the immature enamel matrix of ameloblastic fibro-odontomas and odontomas. Furthermore, enamelysin was also detected in globular amyloid masses and calcified foci in calcifying epithelial odontogenic tumors, hyaline droplets, small and large mineralized areas in adenomatoid odontogenic tumors, and a portion of ghost cells in calcifying odontogenic cysts. Positive reactivity was also observed in selected tumor cells in some of these tumors. No intracellular staining for enamelysin was detected in ameloblastomas or the ameloblastic portion of ameloblastic fibro-odontomas. Also, enamelysin was not detected in dentin, dysplastic dentinoid hyaline matrices, and cementum that were present within the tumors examined. Thus, taken together, our results suggest that the enamelysin-specific monoclonal antibody (203-1C7) may be utilized as a marker of early enamel development and that enamelysin may be involved in the pathogenesis of specific odontogenic tumors.

Descending necrotizing mediastinitis due to odontogenic infections.

OBJECTIVE: Acute purulent mediastinitis caused by oropharyngeal infection is termed descending necrotizing mediastinitis. Such infections usually have a fulminate course, leading to sepsis and frequently to death. The purpose of this study is to show the importance of early diagnosis, aggressive surgical intervention, and optimal antibiotics chemotherapy in controlling this fatal infectious disease. STUDY DESIGN: Two patients with descending necrotizing mediastinitis due to odontogenic infection who were treated at our institution are described. RESULTS: Both patients survived. CONCLUSIONS: From the patients, 23 different aerobic and anaerobic bacteria were isolated. All of the isolates were susceptible to carbapenem. Early evaluation by means of cervicothoracic computed tomography scanning was extremely useful for diagnosis and surgical planning. Knowledge of anatomic pathways from the mouth to the mediastinum is essential. We believe that tracheostomy is not always necessary. In both of the cases presented, mediastinal drainage was completed through use of a transcervical approach. However, a more aggressive drainage including tracheostomy might be necessary when the infection extends below the carina.

Recent Outbreak of Soybean Sudden Death Syndrome Caused by Fusarium virguliforme and F. tucumaniae in Argentina.

Sudden death syndrome (SDS) of soybean was detected initially in Argentina during 1991-1992 in the Pampas Region and 1992-1993 in the Northwest Region. The first report of the fulfillment of Koch's postulates of SDS caused by Fusarium solani f. sp. glycines in Argentina was published in 2003 (3). Subsequently, analyses have shown that F. solani f. sp. glycines represents several morphologically and phylogenetically distinct species, including F. tucumaniae in Argentina and F. virguliforme in the United States (1). Isolations were made from plants that exhibited typical SDS symptoms (interveinal foliar chlorosis and necrosis leading to defoliation of the leaflets but not the petioles) from fields in Santa Fe and Buenos Aires provinces in 2001, 2002, and 2003. To determine which species are responsible for SDS in Argentina, cultures of eight slow growing isolates that developed bluish pigmentation and produced abundant macroconidia in sporodochia on potato dextrose agar were subjected to morphological and molecular phylogenetic analyses and pathogenicity tests. Morphological analyses demonstrated that three of the isolates were F. virguliforme and five were F. tucumaniae. Isolates of F. tucumaniae produced long and narrow sporodochial conidia while F. virguliforme produced diagnostic comma-shaped conidia. Molecular phylogenetic analyses of DNA sequences from multiple loci confirmed morphology-based identifications and showed that the soybean SDS pathogen in the United States, F. virguliforme, was also present in Argentina. To our knowledge, this is the first report of F. virguliforme in Argentina and of this pathogen outside the United States. Five isolates of F. tucumaniae and three isolates of F. virguliforme were used for pathogenicity tests. F. virguliforme isolate 171 provided by J. Rupe (University of Arkansas, Fayetteville) was used as a positive control. Soybean cultivars Ripley, RA 702, Pioneer 9492RR, Spencer, and A-6445RG were inoculated with each of the isolates tested in a greenhouse assay using soil infestation and toothpick methods (2). All eight isolates produced typical foliar SDS symptoms 15 to 25 days after inoculation. Severity of foliar symptoms averaged 3.3 for F. virguliforme, 2.6 for F. tucumaniae, and 3.3 for the positive control using a disease severity scale in which 1 = no symptoms and 5 = severely infected or dead plants. Under these conditions, F. virguliforme appeared to be more virulent than F tucumaniae. Noninoculated plants remained symptomless. Koch's postulates were confirmed with soybean cultivars RA 702 and A6445RG. Isolates recovered from symptomatic plants inoculated by the soil infestation and toothpick methods were identical to those used to inoculate the plant. Strains were recovered at frequencies of 100 and 60% from plants inoculated by the toothpick and soil infestation methods, respectively. To our knowledge, this is the first report of the fulfillment of Koch's postulates for F. tucumaniae and F. virguliforme in Argentina. References: (1) T. Aoki et al. Mycologia 95:660, 2003. (2) K. W. Roy et al. Plant Dis. 81:1100, 1997 (3) M. Scandiani et al. Plant Dis. 87:447, 2003.

Comparison of plastic prostheses and self-expandable metallic stents in the treatment of malignant esophageal stenosis.

OBJECTIVE: We compared the efficacy of plastic prostheses and self-expandable metallic stents in the treatment of malignant esophageal stenosis and/or fistula. SUBJECTS AND METHODS: Subjects were 31 patients with esophageal cancer, 4 with esophagotracheal fistula, and 1 with esophageal stenosis. A plastic prosthesis was inserted in 15 patients (group A) and a self-expandable metallic stent in 21 patients (group B). We evaluated food ingestion improvement, effectiveness, complications, mean survival, food ingestion duration, the percentage of food ingestion capability in total survival time, and inhospital mortality. RESULTS: No differences were seen in food intake improvement or in the effectiveness between groups, whereas fatal complications were higher in Group A. No significant differences were seen in mean survival, food ingestion duration, percentage of food ingestion capability, or inhospital mortality between groups. CONCLUSION: We concluded that a self-expandable metallic stent was safer than a plastic prosthesis because of fewer serious complications such as bleeding, and recommended the use of metallic stents in the treatment of malignant esophageal stenosis and/or fistula.

A case of rupture of the internal jugular vein caused by postoperative infection of functional neck dissection.

A 68-year old diabetic man with gingival cancer of the lower jaw underwent resection of the mandible and functional neck dissection. Swabs of a postoperative wound infection revealed methicillin-resistant Staphylococcus aureus (MRSA). The wound was irrigated, and antibiotics administered. The pathogens isolated were sensitive to the antibiotics used, but the infection failed to respond to treatment. Bleeding ensued on the 14th postoperative day (#14POD), when the wound was opened to reveal thrombosis and rupture of the right internal jugular vein. The patient's condition improved after ligation and surgical debridement of the right internal jugular vein. Our experience underlines the importance of early radiological investigation for possible thrombus formation in the internal jugular vein in cases of postoperative wound infection follows functional neck dissection with conservation of the internal jugular vein. It is also important to actively treat this condition surgically, including ligation of internal jugular vein for suppressing inflammation.

A secure method of nasal endotracheal tube stabilization with suture and rubber tube.

A new method of stabilizing the nasal endotrascheal tube was described. The tube was secured to the anterior portion of the nasal septum with braided silk thread, which was tightened over the rubber tube to keep air route of the cuff open. There found no complications such as unplanned extubation, necrosis and infections of the nasal septum.

Acinic cell carcinoma of the sublingual gland accompanied by bone formation.

A rare case of acinic cell carcinoma of the sublingual gland accompanied by bone formation is reported. The patient is a 79-year-old male who was referred to Yokohama Minami Kyosai Hospital with sublingual swelling. A tumor mass, 20 x 10 mm in diameter, was detected on the right side of the floor of the mouth. Computed tomography (CT) revealed a mass lesion with calcification in the sublingual gland. The patient underwent total sialadenectomy of the sublingual gland with conservation of the lingual nerve. Histologically, the lesion showed amylase-positive atypical cells with thyroid gland-like arrangement, and mature bone tissue in the stroma. Based on these findings, the tumor was diagnosed as acinic cell carcinoma accompanied by bone formation. Postoperative recovery was uneventful, and two years after surgery, there are no signs of distant metastases or recurrence.

Simultaneous treatment of multiple primary cancers of the oral cavity and other sites.

Multiple primary cancers are not uncommon in the head and neck region. Since the time for treatment will be prolonged if each lesion is treated separately, simultaneous treatment of the cancers is preferred to ensure complete remission of lesions and increased survival of patients. In this paper, the efficacy of combined treatment was evaluated in 9 patients with oral cancers and concurrent cancers in other sites. The mean age of the patients was 54 years (range 44 to 66). The tongue (8) and mandible (1) were the sites of involvement. Concurrent cancers were found most often in the esophagus, followed by stomach and lung. Histologically, 7 lesions were diagnosed as squamous cell carcinomas. All oral cancers were treated by surgery. Neck dissection and simultaneous reconstruction were performed in 5 patients. In addition, concurrent cancers were treated simultaneously by endoscopic mucosal resection in 2 patients and radical resection followed by immediate reconstruction in 6 other patients. However, simultaneous surgical treatment of all lesions could not be performed in an individual with 3 cancers. The time of surgery ranged from 23 minutes to 17 hours and 30 minutes. With the exception of 2 patients who died of postoperative complications and had needed treatment for dysphasia, all patients were discharged after 1 to 2 months. Four of the patients are still alive 2 years and 6 months after surgery. These results indicate that simultaneous treatment is effective in selected cases of multiple cancers.