Polyphenols and fatty acids responsible for anti-cyanobacterial allelopathic effects of submerged macrophyte Myriophyllum spicatum.

Myriophyllum spicatum is known to inhibit the growth of cyanobacteria such as Microcystis aeruginosa by releasing anti-cyanobacterial allelochemicals. The allelochemicals possibly responsible for the inhibition include five polyphenols and three fatty acids, but the extent to which these are indeed responsible for the anti-cyanobacterial effects is unclear. The goal of this research was to determine the contribution of these compounds to the allelopathic effect of M. spicatum on M. aeruginosa. We first collected information on the release rates of these compounds and then added the compounds to a cyanobacterial medium on the basis of their release rates so as to simulate their excretion by M. spicatum. Addition of the polyphenols and fatty acids inhibited the growth of M. aeruginosa, and the interaction of the polyphenols and fatty acids was additive. The EC50 of a polyphenol and fatty acid mixture was compared with that of M. spicatum itself as previously determined in a mixed culture system in which M. spicatum and M. aeruginosa were incubated. The former was about 1.9 times higher than that of the latter, the implication being that the inhibitory effect of the polyphenols and fatty acids contributed about 53% of the allelopathic effect of M. spicatum. This paper is the first to describe allelochemicals that account for a half of the anti-cyanobacterial allelopathic effect of a macrophyte.

Atypical Internal Yellowing of Papaya Fruit in Hawaii Caused by Enterobacter sakazakii.

Internal yellowing (IY) caused by Enterobacter cloacae and characterized by yellow discolored tissue surrounding the papaya (Carica papaya L.) seed cavity, diffuse margins, and the presence of a distinctly rotten odor was first reported in 1987 (3). Here we report the formation of atypical internal yellowing (AIY) in ripe papaya caused by the bacterium Enterobacter sakazakii. In surveys conducted from 2006 to 2007, 'Kapoho Solo' papayas grown in the Puna District of Hawaii Island were obtained from various packinghouses. After incubation at 27°C, the papayas were bisected and examined for symptoms of IY. Among papayas that were asymptomatic for IY, a dull, greenish yellow discoloration of the flesh with a distinct margin extending from the seed cavity into the pericarp was noted, along with a pungent odor. These symptoms occurred in 5 of the 500 fruit surveyed and bacterial populations were 102 to 103 CFU/g. Discolored tissue was aseptically excised, weighed, macerated, serially diluted in sterile distilled water (SDW), and plated onto modified peptone yeast extract medium (PT-M4) (4). The plates were incubated at 30°C for 24 to 48 h until single colonies were evident. After 48 h, colonies on PT-M4 were orange-red, convex and circular, and surrounded by a somewhat opaque 1-mm margin. After single colony purification, five strains were obtained. The strains, inoculated into oxidation/fermentation-glucose tubes and API 20E strips (bioMerieux, Inc., Durham, NC) incubated at 30°C, were shown to be facultative anaerobes and identified as E. sakazakii with a 98.4% certainty. Colonies plated onto tryptic soy agar (TSA) and incubated for 72 h at 25°C produced yellow pigmentation, indicative of E. sakazakii. Amplification by PCR with E. sakazakii-specific primers (2) yielded a 929-bp fragment, which was absent with E. cloacae and Pseudomonas aeruginosa template DNA. To confirm pathogenicity, cell suspensions at 109 CFU/ml of putative E. sakazakii strains RK07-05, RK07-06, and RK07-07 and E. cloacae (3) were inoculated by injection (0.5 ml per site) into one-third-ripe 'Kapoho Solo' papayas (six fruit per strain, inoculated at duplicate sites) and incubated at 27°C for 4 days. Control sites were injected with 0.5 ml of SDW. Fruit inoculation experiments were repeated. E. cloacae-inoculated sites produced typical IY as previously described (3), while the sites inoculated with the three E. sakazakii strains produced greenish yellow tissue (26% mean incidence), symptomatic of AIY. Control sites did not produce IY or AIY. Koch's postulates were fulfilled, and the identification of reisolated bacterial strains was confirmed with API 20E, PCR, and pigment production on TSA. Although less prevalent (1% incidence) than the typical IY produced by E. cloacae (3), E. sakazakii has the potential to affect quality and food safety of fresh and processed papaya products. E. sakazakii has been implicated in a severe form of neonatal meningitis, sepsis, and necrotizing enterocolitis (1). Research into the transmission and infection of papaya of this cross-domain pathogen merits further study. References: (1) D. H. Adamson. Clin. Microbiol. Newsl. 3:19, 1981. (2) A. Lehner et al. BMC Microbiol. 4:43, 2004. (3) K. A. Nishijima et al. Plant Dis. 71:1029, 1987. (4) K. A. Nishijima et al. Plant Dis. 88:1318, 2004.

Organic carbon leaching behavior from incinerator bottom ash.

The understanding of the leaching behavior of organic carbon from incinerator bottom ash is an important aspect for the control of organic carbon emissions from landfills in order to minimize their potential risk to the environment. The leaching behavior of organic carbon from incinerator bottom ash samples, obtained from two different solid waste sources, as well as the effects of calcium (Ca) on organic carbon (DOC) leaching associated with pH were therefore investigated in this paper. The effect of pH on the leaching of DOC and other major elements from bottom ash was ascertained through pH-stat leaching experiments. Column leaching experiments were carried out to evaluate the dependence of the leached amount of DOC on Ca leaching. It was found that the bottom ash produced by incineration of municipal solid waste (MSW) was composed of two DOC fractions: DOC leached independent (fraction I) of Ca leaching, observed at alkaline-neutral pH, and DOC leached dependent (fraction II) on Ca leaching, observed at acid pH. However, the bottom ash produced by incineration of industrial solid waste (ISW) was composed of only DOC fraction I. The addition of calcium oxide during incineration and the differences in the paper to plastic ratio in the physical composition of the solid wastes incinerated would explain the distinct organic carbon leaching behaviors of bottom ash samples.

Chemical extraction of organic carbon to reduce the leaching potential risk from MSWI bottom ash.

The performance of extraction solvents, including organic and inorganic solvents, for organic carbon extraction from municipal solid waste incinerator (MSWI) bottom ash was evaluated. The total carbon (TC) extracted was used to ascertain the efficiency of extraction solvents and the reduction of dissolved organic carbon (DOC) leaching potential was used to evaluate the capacity of solvents to minimize environmental impacts of MSWI bottom ash over short- and long-term considerations in landfill sites. Extract final pH value was a prominent parameter affecting TC extraction. The higher efficiency was obtained at the lower extract final pH and acid or neutral condition was necessary to achieve approximately 30% of TC extraction from bottom ash. On the basis of the results of TC extraction, the efficiency of organic carbon reduction was evaluated using organic carbon leaching potential. Hydrochloric acid was the best solvent to extract organic carbon in controlled pH conditions. Hydrochloric acid reduced the organic carbon leaching potential of MSWI bottom ash by about 68% at neutral leaching pH.

Adenocarcinoma of the sublingual gland: case report and 50 year review of the literature.

Primary adenocarcinoma arising in the sublingual gland is extremely rara, accounting for six cases reported in extensive survey of the literature for the period of 1930 to 1981. A case of adenocarcinoma of the sublingual gland is documented, demonstrating a highly malignant nature of the disease. This neoplasm possesses particular therapeutic problems owing to the relative lack of symptoms and the delay of initial medical observation after the onset of symptoms.

Analyses of distant metastases in squamous cell carcinoma of the head and neck and lesions above the clavicle at autopsy.

The frequency of distant metastases was studied in 112 patients who had squamous cell carcinoma of the head and neck. All of these patients died of the tumor and underwent postmortem examinations. Forty-one (37%) of the 112 patients had distant metastases at the time of death. There was a significant correlation between development of distant metastases and the presence of cervical nodes. Of the patients who had distant metastases, 63% had evidence of tumor in the cervical nodes, whereas 37% of these patients were free of disease in the neck. Patients with residual or recurrent tumors in the cervical nodes had a high risk of distant metastases.

Coagulation mechanism of salt solution-extracted active component in Moringa oleifera seeds.

This study focuses on the coagulation mechanism by the purified coagulant solution (MOC-SC-PC) with the coagulation active component extracted from M. oleifera seeds using salt solution. The addition of MOC-SC-PC tap water formed insoluble matters. This formation was responsible for kaolin coagulation. On the other hand, insoluble matters were not formed when the MOC-SC-PC was added into distilled water. The formation was affected by Ca2+ or other bivalent cations which may connect each molecule of the active coagulation component in MOC-SC-PC and form a net-like structure. The coagulation mechanism of MOC-SC-PC seemed to be an enmeshment of Kaolin by the insoluble matters with the net-like structure. In case of Ca2+ ion (bivalent cations), at least 0.2 mM was necessary for coagulation at 0.3 mgC l-1 dose of MOC-SC-PC. Other coagulation mechanisms like compression of double layer, interparticle bridging or charge neutralization were not responsible for the coagulation by MOC-SC-PC.

Isolation and characterization of coagulant extracted from Moringa oleifera seed by salt solution.

It is known that M. oleifera contains a natural coagulant in the seeds. In our previous research, the method using salt water to extract the active coagulation component from M. oleifera seeds was developed and compared with the conventional method using water. In this research, the active coagulation component was purified from a NaCl solution crude extract of Moringa oleifera seeds. The active component was isolated and purified from the crude extract through a sequence of steps that included salting-out by dialysis, removal of lipids and carbohydrates by homogenization with acetone, and anion exchange. Specific coagulation activity of the active material increased up to 34 times more than the crude extract after the ion exchange. The active component was not the same as that of water extract. The molecular weight was about 3000 Da. The Lowry method and the phenol-sulfuric acid method indicated that the active component was neither protein nor polysaccharide. The optimum pH of the purified active component for coagulation of turbidity was pH 8 and above. Different from the conventional water extracts, the active component can be used for waters with low turbidity without increase in the dissolved organic carbon concentration.

A new procedure to produce lignocellulosic anion exchangers from agricultural waste materials.

Two lignocellulosic agricultural waste materials (LCM), sugarcane bagasse (BG) and rice hull (RH), were converted into weak-base anion exchanger and evaluated for their exchanger capacity for nitrate. Pure cellulose (PC) and pure alkaline lignin (PL) were also used as reference materials to elucidate possible reactivity in LCM. Epoxy and amino groups were introduced into BG, RH, PC and PL substrates after the reaction with epichlorohydrin and dimethylamine in the presence of pyridine and an organic solvent N,N-dimethylformamide (DMF). Amino group incorporation into cellulose decreased with the presence of water in the reaction mixture and increased with the reaction time and presence of a catalyst (pyridine). The highest maximum nitrate exchange capacity (Qmax) and yields of the prepared exchangers was obtained from PL (1.8 mmol g(-1) and 412.5%), followed by BG (1.41 mmol g(-1) and 300%), PC (1.34 mmol g(-1) and 166%) and RH (1.32 mmol g(-1) and 180%). The proposed synthetic procedure was effective in modifying PL, PC and LCM chemically resulting in a higher yield and nitrate removal capacity.

Preparation of agricultural residue anion exchangers and its nitrate maximum adsorption capacity.

Anion exchangers were prepared from different agricultural residues (AR) after reaction with epichlorohydrin and dimethylamine in the presence of pyridine and N,N-dimethylformamide (EDM method). Agricultural residues anion exchangers (AR-AE) produced by the EDM method were inexpensive and showed almost the same NO3- removal capacities as Amberlite IRA-900. AR-AE produced from AR with higher hemicelluloses, lignin, ash and extractive contents resulted in the lower yields. Sugarcane bagasse with the highest alpha-cellulose contents of 51.2% had the highest yield (225%) and lowest preparation cost. The highest maximum adsorption capacity (Qmax) for nitrate was obtained from rice hull (1.21 mmol g(-1)) and pine bark natural exchangers (1.06 mmol g(-1)). No correlation was found between Qmax and alpha-cellulose content in the original AR. AR-AE produced from different AR demonstrated comparable Qmax due to the removal of non-active compounds such as extractives, lignin and hemicelluloses from AR during the preparation process. Similar preparation from pure cellulose and pure alkaline lignin demonstrated that the EDM method could not produce anion exchangers from pure lignin due to its solubilization after the reaction with epichlorohydrin.

Primary chondroid chordoma arising from the base of the temporal bone. A 10-year post-operative follow-up.

A 36-year-old male with a primary chondroid chordoma is presented. This tumour arose from the base of the temporal bone and extended to the mastoid cavity. It involved the facial nerve and was adherent to the internal jugular vein and internal carotid artery. The tumour was excised and the patient has been carefully followed up for 10 years. He has shown no evidence of local recurrence, intracranial extension of the residual tumour and distant metastasis.

Numerical modelling of the petroleum oil penetration into sandy beach sediments.

A numerical model describing the penetration of petroleum oil into sandy beach sediments was developed to assess the behavior of stranded oil at tidal zone as a result of tanker accidents, and so on. To understand the penetration behavior, penetration rate of three species of petroleum oil (two species of fuel oil C and one species of crude oil) was observed experimentally with artificial tidal zone equipment. As a consequence, two types of oil were distinguished from the viewpoint of penetration rate. One (fuel oil C-1) kept its homogeneity in composition and showed relatively rapid penetration, the other (fuel oil C-2 and crude oil) became heterogeneous and slow in penetration. Considering this aspect as the adsorption of polar compounds (i.e. asphaltenes) on the sediment surface, a numerical model that described oil penetration into sandy beach was developed. As a result, the difference in penetration rate between these two types of oil could not be replicated sufficiently only by consideration of the adsorption. However, the change of a parameter value which represents the apparent viscosity of oil led to good agreement with observations. Simulation results indicated that when fuel oil C or crude oil used in this study was stranded at a sandy beach located in Hiroshima Bay, Japan, 2 to 39% of total stranded oil might penetrate into the deeper zone (> 3 cm in depth) over 50 days.

Stimulation of bacterial activity by the addition of different PACs.

The effect of bacterial adhesion on powdered activated carbon on bacterial activity in non-toxic wastewater was studied and the difference of powdered activated carbon species on the stimulation of bacterial activity was also evaluated using eight kinds of powdered activated carbon. Pure cultures of Escherichia coli K-12, which can reduce NO3- to NO2- in anoxic conditions, were used. Specific nitrate reduction activity was used as an indicator of bacterial activity. The higher specific nitrate reduction rates were noted in cultures with powdered activated carbon than without powdered activated carbon or with Kaolin. The specific nitrate reduction rate with powdered activated carbon was up to about 1.4 times as highas those without powdered activated carbon. The stimulation of bacterial activity was induced by the adhesion of bacterial cells onto powdered activated carbon. There are powderedactivated carbons that can stimulate bacterial activity, whereas not all the powdered activated carbon can stimulate bacterial activity. Surface characteristics like Brunauer-Emmett-Teller specific surface areas, total surface acidity, oxygen functional groups, total surface basicity, surface charge, pH(pzc), iodine number, concentrations of metals, electric resistance, free radical concentration and adsorption capacity of formate did not related with the stimulation of the bacterial activity.

[High dose methotrexate with CF rescue therapy in patients with head and neck cancer].

A clinical trial of high dose methotrexate (MTX)-CF rescue was conducted in 17 institutions. Forty-seven patients with head and neck cancer entered this trial, of which 29 were evaluable. In this series, the patients were divided into 2 groups (Arm I, Arm II), according to physician's selection. MTX was administered 700 mg/m2 (1000 mg/body) in Arm I and 1750 mg/m2 (2500 mg/body) in Arm II by i.v. drip over 6 hours. Twenty-four hours after initiation of MTX infusion, CF rescue was started. There was no complete response in 29 patients. Four patients showed partial response with an overall response rate of 13.8%. There were 4 partial responders out of 21 patients in Arm I, with a response rate of 19%, whereas there was no partial responders out of 8 patients in Arm II. These results showed no apparent dose response. MTX concentrations in plasma were determined at 6, 24, 48 and 72 hours after initiation of MTX infusion. The assay results revealed a safe range in Arm I, but exceeded in 3 cases of Arm II. GI disturbances were seen at the rate of 78%. Bone marrow suppression was remarkable and hepatic toxicity was observed as the rate of 41%. No renal toxicity was observed. The results of high dose MTX-CF rescue therapy were not better than those of moderate dose therapy, so that we concluded that MTX should be used as one agent at low or moderate dose in combination chemotherapy as far as Japanese patients with head and neck cancer are concerned.

[Moderate dose methotrexate with citrovorum factor rescue therapy in the treatment of head and neck cancer].

A clinical trial of moderate dose methotrexate (MTX)-CF rescue was conducted in 12 institutions. Thirty-seven patients with head and neck carcinoma entered this trial, of which 32 were evaluable. MTX was administered 350 mg/m2 (500 mg/body) by i.v. drip over 6 hours. Three hours after completion of MTX infusion, CF rescue was started. There was no complete response in 32 patients. Nine patients showed partial response with the response rate of 28%. The response rates were 21% for the group of patients treated previously, and 75% for the group untreated previously. MTX concentration in plasma was determined at 6, 24, 48 and 72 hours after the initiation of MTX infusion, and the assay results revealed a safe range. GI disturbances were seen at the rates of 11 to 38%. Bone marrow suppression was mild and no renal toxicity was observed. We concluded that the moderate dose MTX-CF rescue therapy was useful for head and neck carcinoma. As a next step, we are planning to conduct a clinical trial of high-dose MTX.

[Simultaneous bilateral neck dissections].

From April 1985 to December 1989, 65 patients with advanced head and neck squamous cell carcinoma, underwent simultaneous bilateral neck dissection (SBND) at Saitama Cancer Center. Three and five year survival percentages were 53 and 42%, respectively. In patients without histologic involvement of cervical nodes, five year survival rate was 83%, whereas in those with nodal involvement five year survival fell to 32% (p less than 0.005). The conclusion were the following: (1) Of 38 patients diagnosed to have lymph node involvements on one side of neck before operation, 8 patients (22%) were found to have bilateral lymph node metastasis in clinicopathological study. Of 13 patients having no clinical lymph node metastasis on both sides of neck, 7 patients (54%) were found to have unilateral lymph node metastasis. Of 16 patients diagnosed to have bilateral lymph nodes involvement, 10 patients were found to have bilateral neck metastasis and 2 had unilateral neck metastasis. (2) Of 35 cases of hypopharyngeal canners, 19 cases had clinically positive lymph nodes on one side of neck. Of these 19 cases, 5 cases (26%) had histologically positive nodes on the opposite side. 14 (40%) of 35 cases had metastasis on the opposite side. In conclusion, SBND is a proper treatment for metastatic cervical cancer from a primary lesion of the head and neck, especially in hypopharyngeal cancers, because the rate of recurrence seems to be related more to the difficulty in controlling lymph node metastasis than to the failure in treatment of the primary cancer.

[Salivary duct carcinoma of the parotid gland. Case report and review of the literature].

Salivary duct carcinoma is an uncommon neoplasm which was first described by Kleinsasser et al. in 1968. They pointed out the histologic resemblance of this tumor to ductal carcinoma of the breast and termed it salivary duct carcinoma. Since then, about sixty cases, including some with suspicious pathologic pictures, have been reported. The characteristic microscopic features of salivary duct carcinoma are composed of comedo necrosis, a cribriform and papillary pattern of intraductal growth and aggressive infiltration to adjacent structures. Salivary duct carcinoma, which is the highest-grade malignancy among salivary gland tumors, must be treated aggressively with curative extended surgery. We present the case of a 74-year-old woman with salivary duct carcinoma of the parotid gland. She complained of left facial palsy and left subaural mass. For this advanced tumor involving facial palsy and subaural cutaneous invasion, en block resection with cutaneous reconstruction with a deltopectoral flap and postoperative radiation therapy was performed. She is alive without evidence of recurrence more than four years after this combined therapy.

[Inverted papillomas in the nose and paranasal sinuses].

Inverted papillomas in the nose and/or paranasal sinuses exhibit a high recurrence rate, and an association with malignancy. Early diagnosis and aggressive surgical therapy are thus essential. Seventeen cases of inverted papilloma seen at Saitama Cancer Center over a 17-year period were reviewed. Common presenting symptoms, the primary papilloma sites and the results of surgical treatment were as follows. 1) Almost all patients complained of nasal obstruction. The usefulness of nasal biopsy of the tumor was confirmed, with 12 cases being diagnosed as having inverted papilloma pre-operatively. Inverted papilloma without squamous cell carcinoma caused osseous thinning, but did not destroy the bone. 2) It was found that the primary site of the papilloma involved the lateral wall of the nasal cavity. Lateral Rhinotomy was therefore recommended as a standard treatment. 3) The recurrence rate was 1/12 after Lateral Rhinotomy. Two cases had complaints associated with the Lateral Rhinotomy, nasolacrimal duct stenosis, and a scar in the median corner of eye. 4) Only one case had concomitant squamous cell carcinoma in the nose and maxillary sinus. This patient received chemo therapy, radiation therapy and finally maxillectomy, but the inverted papilloma recurred several times. Six years later, squamous cell carcinoma recurred and lead to this patient's death.

[A case of rhinosporidiosis].

Rhinosporidiosis is due to fungal infection by Rhinosporidium seeberi, which affects predominantly the mucous membrane of the nose and nasopharynx. This disease is characterized by the formation of papillomatous and polypoid lesions and is known to be endemic in India and Sri Lanka. The first case of rhinosporidiosis in Japan is reported in this paper. A 25-year-old male Indian, born in India, living in Japan for a year, visited the outpatient clinic complaining of nasal obstruction and swallowing pain. A friable, irregular, reddish polypoid mass was found in the right nasal cavity. Endoscopy as well as plain X-ray and CT examination showed that the mass originated from the right inferior turbinate and extended into the choana without any sign of bone destruction. The mass removed under general anesthesia showed the typical feature of rhinosporidiosis, that is, papillomatous hyperplasia of the mucosa with sporangia full of spores in different stages of development. One year after treatment, no relapse has been seen in this case.

[Reconstruction of hypopharynx and cervical esophagus using a free jejunal graft].

Forty-four patients with cancer in the hypopharynx and cervical esophagus underwent pharyngoesophageal reconstruction with a free jejunal graft after total removal of a tumor combined with resection of the circumferential tissues. The graft survival rate was 93.2% (41 of 44). Postoperative complications occurred in 16 patients (36.4%), which included postoperative death in one (2.3%), graft necrosis in 3 (6.8%), fistula formation in 4 (9.1%) and graft stricture in 3 (6.8%). All 3 cases of graft necrosis underwent successfully reimplantation. The mean interval time was 20.9 days until oral intake was possible. Thirty-two of 42 patients (76.2%) were able to take usual food. Esophageal speech was limited in the selected cases by this procedure. A cumulative survival rate of 5 years was 40.6% in this series. This method of reconstruction should be advocated as one of relatively safe and reliable procedures for patients with hypopharyngeal and cervical esophageal cancer.