A Simple Method for Separating Phenanthrene from Soil by Cloud-Point Extraction

Contamination of soil by polycyclic aromatic hydrocarbons (PAHs) is currently widespread in urban and industrial areas, and the decontamination of PAHs remains a challenge. In addition, recovering PAHs from large volumes of soil washings is costly. Therefore, in this study, we focused on cloud-point extraction (CPE) without centrifugation, which separates PAHs from the washing solution through gravitational sedimentation. Specifically, we examined the conditions for the separation of phenanthrene (a typical PAH pollutant) from soil using CPE. After evaluating the water and phenanthrene solubilities of 23 commercially available nonionic surfactants, Brij 30 was found to be optimal. We simulated contaminated soil by adding phenanthrene to three soils with different particle sizes and varying amounts of organic matter. We examined the Brij 30 washing conditions and the effects of salt additives that promote phase separation during CPE. The addition of either sodium chloride or sodium sulfate enabled CPE at 25°C, but sodium sulfate was found to be more effective at lower concentrations than sodium chloride. A phenanthrene recovery rate of 58–88% was observed for each laboratory-simulated contaminated soil by CPE using salt additives. This method is economical and effective for processing large amounts of contaminated soil.     

Peritoneal washing cytology in cervical carcinoma. Analysis of 109 patients

The peritoneal washing cytologies of 109 patients (112 procedures) undergoing laparotomy for cervical carcinoma were evaluated retrospectively and compared with the clinical and pathologic findings. Nine patients (8.3%) had malignant peritoneal washings (including three of four with washings initially termed "inconclusive"). Four (4.9%) of the 82 patients with squamous carcinoma and 3 (16.7%) of 18 with adenocarcinoma had positive washings. Five (5.6%) of 90 washings obtained at initial explorations were positive, as compared with 4 (18.2%) of 22 washings obtained as follow-up operations in recurrent cases. The 111 peritoneal washing cytologies with a corresponding histologic evaluation of the peritoneal cavity showed a good correlation; peritoneal washing cytology had an efficiency of 91.0%, a sensitivity of 52.9% and a specificity of 100%. Two cases in which the cytologies were considered positive only after review had negative peritoneal histologies; both patients died of progressive disease within 11 months. Peritoneal washing cytology was positive in 5 (5.9%) of 84 cases with FIGO stage 1 cancers, 2 (18.2%) of 11 cases with stage 2 cancers, 1 (33.3%) of 3 cases with stage 3 cancers, and 1 (10%) of 10 cases with recurrent tumors. Eight (88.9%) of nine patients with malignant peritoneal washings died of disease from 3 to 15 months following surgery; one showed no evidence of disease at 9 months. These results suggest that: (1) cervical carcinomas are infrequently associated with a positive peritoneal washing; (2) peritoneal washing cytology is more likely to be positive in cases of adenocarcinoma than in cases of squamous carcinoma; (3) peritoneal washings obtained at the time of surgery for recurrence are more likely to contain malignant cells than are washings obtained during initial exploration; (4) nonkeratinizing malignant squamous cells may be confused with reactive mesothelial cells; and (5) peritoneal washing cytology is a relatively insensitive technique for detecting advanced cervical disease, but correlates with a poor prognosis when positive.     

Cytologic evaluation of low grade transitional cell carcinoma and instrument artifact in bladder washings

OBJECTIVE: To identify key cytologic features for the separation of low grade transitional cell carcinomas (TCCs) from nonneoplastic lesions in bladder washings. STUDY DESIGN: The cytomorphologic features of 95 bladder washing specimens showing papillary fragments, which included 50 low grade TCCs and 45 nonneoplastic lesions, were reviewed retrospectively. RESULTS: Bladder washings from low grade TCCs showed papillary and irregular groups of cells with ragged borders, cytoplasmic homogeneity and subtle nuclear changes, such as increased nuclear/cytoplasmic ratio and irregular nuclear border. Bladder washings after instrumentation from nonneoplastic lesions of the bladder showed cellular specimens with cohesive, ball-shaped and papillary clusters with smooth borders lined with a denser-staining cytoplasmic collar. Reactive urothelial cells often displayed loose aggregates with irregular borders but no cytoplasmic collar. CONCLUSION: In bladder washing cytology, nuclear changes and cytoplasmic homogeneity play a major role in the diagnosis of carcinoma.     

Studies on the influence of leaf washings on infection by Mycosphaerella ligulicola

Infection of chrysanthemum leaf discs by spores of Mycosphaerella ligulicola was increased in the presence of concentrated leaf washings obtained from five plant species tested. On separation of leaf washings by dialysis the residue stimulated infection and caused aggregation of hyphae on leaf and cellophane surfaces. In the presence of the diffusible fraction an extensive branching network of hyphae developed but there was no increase in infection except in the presence of the chrysanthemum diffusate. Infection of leaf discs of certain plants normally resistant to M. ligulicola occurred in the presence of concentrated leaf washings. Leaf washings from a resistant variety of chrysanthemum had the same effects as those from a susceptible variety. An extract of chrysanthemum leaves was less effective in stimulating infection than was a leaf washing. Defined nutrient media and potato extract stimulated infection but caused the development of different growth habits on leaf surfaces. The active components of the dialysis residue of chrysanthemum leaf washings were heat-stable but apparently volatile. Eluates from strips of certain regions of paper chromatographs of leaf washings and potato extract caused increased infection together with development of characteristic growth habits of the fungus on the leaf.     

Biosurfactant Mediated Remediation Process Evaluation on a Mixture of Heavy Metal Spiked Topsoil Using Soil Column and Batch Washing Methods

This study investigated the effects of biosurfactant produced by a mangrove isolate on a heavy metal spiked soil remediation using two different methods of biosurfactant addition (pretreatment and direct application) at different concentrations (0.5%–5%) for 10 days employing column and batch method of washings. The FT-IR spectral and biochemical analysis confirmed the chemical nature of biosurfactant as a glycolipid. Pre-addition of biosurfactant at 0.5% concentrations and further incubation for a month resulted in better chromium removal than the direct biosurfactant washing method. A maximum recovery of lead (99.77%), nickel (98.23%), copper (99.62%), and cadmium (99.71%) were achieved with column washing method at 1% biosurfactant concentration. Release of 26% soluble fractions of nickel (pre-addition with biosurfactant) and 40% copper (direct application) were achieved by column washing method at 1.0% concentration of biosurfactant. A total of 0.034 mg/10 g of lead, 0.157 mg/10 g of nickel, 0.022 mg/10 g of copper, 0.025 mg/10 g of cadmium, and 0.538 mg/10 g of chromium were found to remain in the spiked soil after column washing with 1.0% biosurfactant solution. However, pre-addition of 0.5% biosurfactant treatment helps in maximum removal of chromium metal leaving a residual concentration of 0.426 mg/10 g of soil, suggesting effective removal at very low concentration. The average extraction concentration of metals in batch washings was between 93–100%, irrespective of the concentration of biosurfactant studied. In this study, the percentage removal of copper, cadmium, chromium, nickel, and lead from spiked soils by column washing was comparatively lower than batch washing.     

Beiträge zur Anwendung der Bodenwaschtechnik für die Isolierung von Bodenpilzen

Zusammenfassung Zwei zeitsparende Modifikationen der Parkinson & Williams-Bodenwaschtechnik für die Isolierung von Pilzen auf Bodenpartikeln werden beschrieben. Das wesentliche Merkmal beider Modelle ist der kontinuierliche Wasserdurchfluß durch das Waschgefäß, das von Hand oder mechanisch geschüttelt werden kann.Die Wirksamkeit der Bodenwaschung in Abhängigkeit von der verwendeten Wassermenge wurde indirekt ermittelt durch Zählung des Sporengehalts im Waschwasser und durch Bestimmung des Kolonisationsquotienten für die gewaschenen Partikel. Wenn der Sporengehalt des Waschwassers auf ein bestimmtes niedriges Niveau abgesunken war, blieb auch der Kolonisationsquotient konstant, unabhängig von der verwendeten Technik.Die Isolierungsfrequenz der häufigsten Pilzarten in Bodenproben derselben Herkunft ließ sich zufriedenstellend reproduzieren. Jahreszeitliche Schwankungen in der Pilzflora auf gewaschenen Partikeln waren unbedeutend. Die Anzahl der Pilze auf Bodenpartikeln nahm mit der Bodentiefe stark ab.Mehr Arten wurden von gewaschenen Partikeln isoliert als aus dem Waschwasser derselben Bodenprobe. Einige Arten, wie Fusarium culmorum, F. avenaceum, F. equiseti, Mucor hiemalis, Ophiobolus graminis, wurden häufiger von organischen als von Mineralpartikeln isoliert; dagegen ließ sich in der Verteilung der meisten von ca. 300 weiteren Arten kein signifikanter Unterschied für die Partikelarten nachweisen. Nur wenige sehr langsamwüchsige Arten wurden ausschließlich von Mineralpartikeln isoliert.

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Contributions to the soil washing technique for the isolation of soil fungi

Summary Two time-saving modifications of the Parkinson and Williams soil washing technique for the isolation of fungi from soil particles are described. The characteristic feature of both modifications is the continuous flow of water through the washing boxes, which can be shaken by hand or mechanically during the washing procedure.The effectiveness of the soil washing depending on the amount of water used was tested indirectly by counting the spore content of the washing water and by determining the colonisation ratio of the washed particles. When the spore content of the washing water had dropped to a certain low level, the number of fungi per washed particle was found to be constant, independently from the technique used.The isolation frequency of the most abundant fungal species in soil samples of the same origin could be reproduced satisfactorily. Seasonal variations in the fungal lora of washed particles were insignificant. The number of fungi colonizing soil particles decreased strongly with soil depth.More species were isolated from washed particles than from dilution plates poured with the washing water of the same soil sample. Some species, e.g. Fusarium culmorum, F. avenaceum, F. equiseti, Mucor hiemalis, Ophiobolus graminis, were more frequent colonizers of organic soil particles, whereas the distribution pattern of about 300 other species did not differ significantly on mineral or organic particles. Only a few very slow growing species were isolated exclusively from mineral particles.

     

Methods for Examining the Microflora of Mouldy Hay

A comparison of four methods of estimating the microflora of mouldy hay showed that more micro-organisms were removed by washing samples than by blowing in a small wind tunnel. By contrast, actinomycete spores were estimated more efficiently from air using an Anderson Sampler than by plating diluted washings because fewer bacteria were blown than washed off the hay. Estimates of fungal spores were larger from counts in haemocytometer cells than by dilution plating but counts of actinomycetes and bacteria were higher by dilution plating. Fungi grew equally well from air or water suspensions and with the Cascade Impactor and Anderson Sampler their numbers were estimated with less variation than for the numbers of 'actinomycetes and bacteria'even on improved media and after correcting for viability, clumping and interference between colonies.     

A simplified assay of enzymes catalyzing ATP-pyrophosphate exchange reactions.

The ATP-pyrophosphate exchange reaction is a widely used method for determination of the activity of enzymes forming enzyme-adenylate intermediates in the course of their catalytic action (1–4). For separation of the labeled ATP formed in the enzyme-catalyzed reaction from the labeled pyrophosphate, the adsorption of ATP on activated Norit is generally used. This method, however, is rather time consuming (considering the extensive washings required for the removal of pyrophosphate from Norit), and its reproducibility is highly dependent on pretreatment of charcoal with acid, Another method of separation applies anion-exchange paper (5). According to our experience, separation on DEAE-cellulose paper (Whatman DE 81) is not sufficient.     

Study of stringency conditions for human papillomavirus DNA detection on cell lines, frozen and paraffin-embedded tissue sections by in situ hybridization with biotinylated probes

In situ hybridization was mainly used for typing human papillomavirus (HPV) in paraffin-embedded or frozen sections under stringent conditions (SC). We tested 5 different conditions of stringency with biotinylated HPV 1, 2, 16 and 18 probes on 3 cell lines (Sihà and CaSki with HPV16, HeLa with HPV18) by varying the concentration of formamide in the hybridization mixture and washings in order to determine the stringency conditions to be used to assess the presence of HPV and its typing: A-low stringency, hybridization at 35 degrees C below the melting temperature of DNA (Tm-35 degrees C) and washings without formamide; B-low stringency, hybridization and washings at Tm-35 degrees C; C-medium stringency, hybridization at Tm-35 degrees C and washings at Tm-12 degrees C; D-high stringency, hybridization at Tm-12 degrees C and washing without formamide; E-very high stringency, hybridization and washings at -12 degrees C. This study showed that HPV typing required a high stringency. On the contrary, under non stringent conditions (NSC), each cell line was positive with the heterologous probes. When 3 to 5 stringency conditions were assayed on 4 frozen samples, similar results were obtained. Typing required high stringency conditions whereas NSC allowed HPV detection. Furthermore, this study demonstrated the specificity of the reaction in lesions positive with more than one type. Stringent (Tm-12 degrees C) and non stringent (Tm-35 degrees C) conditions of hybridization were further applied to 57 biopsy sections (17 frozen and 40 paraffin-embedded specimens) from typical wart lesions and lesions suspected of HPV.(ABSTRACT TRUNCATED AT 250 WORDS)     

Biodiesel production using heterogeneous catalysts

The production and use of biodiesel has seen a quantum jump in the recent past due to benefits associated with its ability to mitigate greenhouse gas (GHG). There are large number of commercial plants producing biodiesel by transesterification of vegetable oils and fats based on base catalyzed (caustic) homogeneous transesterification of oils. However, homogeneous process needs steps of glycerol separation, washings, very stringent and extremely low limits of Na, K, glycerides and moisture limits in biodiesel. Heterogeneous catalyzed production of biodiesel has emerged as a preferred route as it is environmentally benign needs no water washing and product separation is much easier. The present report is review of the progress made in development of heterogeneous catalysts suitable for biodiesel production. This review shall help in selection of suitable catalysts and the optimum conditions for biodiesel production.     

Cytohistologic correlation of peritoneal washing cytology in gynecologic disease

The peritoneal washings and cul de sac aspirates from 204 patients undergoing 217 procedures for the evaluation of gynecologic disease were examined retrospectively and correlated with the histologic diagnoses. Of the 73 washings from patients with histologically benign genital disease, cytology diagnosed 64 (87.7%) as negative, 6 (8.2%) as inconclusive and 3 as malignant. One malignant washing was a true positive from a nongenital primary. False positives thus occurred in 2.7% of the benign cases on blind review. Of 144 cytologic examinations of washings from patients with histologically confirmed malignant disease of the female genital tract, 38 (26.4%) were considered positive after cytohistologic correlation. Four malignant cases (2.1%) were undercalled on blind review while 3 (2.0%) were considered overcalls. Eleven of 47 cases (23.4%) with biopsy-proven peritoneal disease had negative cytology after histologic correlation. Recurring problems in interpreting peritoneal washings included: (1) the differential diagnosis of the spectrum encompassed by reactive mesothelium, endosalpingiosis, borderline serous tumors and well-differentiated serous cystadenocarcinoma; (2) morphologic similarities between some tumor cells and mesothelial cells associated with treatment effects; and (3) a paucity of malignant cells in some washings, resulting in false-negative interpretations. Ineffective cytopreparation, particularly of bloody specimens, hampered interpretation of some specimens. Correlation with previous histology and cytology enhanced the accuracy of peritoneal washing cytology in this study.     

An automatic soil-washing apparatus for fungal isolation

Summary An apparatus for serial washing of soil particles is described. When forest humus was washed 50 times in an illustrative experiment, the proportion of dark and slow-growing mycelia among fungal isolates was increased from 5 to over 50 per cent.     

Müllerian inclusions in peritoneal washings. Potential source of error in cytologic diagnosis

Müllerian inclusions in peritoneal washings from female patients may be mistaken for adenocarcinoma. Such findings were studied in the peritoneal washing cytology specimens from eight cases. The inclusions usually presented as tubular or papillary structures, often forming a single layer of epithelium surrounding psammoma bodies. The cells forming these structures often displayed some degree of atypia. Recognition of this entity in peritoneal fluids is important to avoid a misdiagnosis of disseminated cancer. A general outline is proposed for interpreting such findings in peritoneal washings, based on the cytomorphology of these structures as well as the microscopic features of the primary neoplasms.     

Despite Long-Term Compost Amendment Seasonal Changes are Main Drivers of Soil Fungal and Bacterial Population Dynamics in a Tuscan Vineyard

The effect of long-term (8 years) compost treatments (compost or compost plus mineral fertilizer) on genetic structure of bacterial and fungal populations in both bulk soil and rhizosphere of grapevine (Vitis vinifera) was analyzed in respect to a control constituted by the soil treated with mineral fertilization. Soils were sampled in early summer (July), mid-summer (August), and before harvest (October). Bacterial and fungal populations were characterized by genetic fingerprints generated by the application of 16S rDNA and ITS rDNA Multiplex Terminal Fragment Length Polymorphism (M-TRFLP) technique. Compost induced no significant differences at any time on microbial communities from bulk soil samples, whereas seasonal variations significantly affected both bacterial and fungal populations as indicated by the Multi Dimensional Scaling (MDS) ordination method of the M-TRFLPs results. MDS analysis of grapevine rhizosphere M-TRFLPs showed that temporal separation was significant for the bacterial population only. Results suggested that soil microbial populations in vineyard productive ecosystems may be sensitive to environmental changes induced by seasonal variations and show a certain degree of resilience to different agricultural practices.     

Soil washing improves the recovery of total community DNA from polluted and high organic content sediments

Treatment of soil with surfactants and chelating agents is used in bioremediation studies to desorb and solubilize contaminants to increase their bioavalability to microorganisms. In the same way that pollutants are made more bioavailable to microorganisms, the procedure can be used to remove potential interfering materials from soil prior to cell lysis and extraction of DNA from indigenous microorganisms. The effect of soil washing was evaluated by extracting DNA from sediments of an intertidal freshwater wetland contaminated with hydrocarbons and from highly contaminated marine sediments from Sydney Harbour, Nova Scotia, Canada. Sediment samples had total organic carbon (TOC) contents that varied between 0.2% and 13%. The chemical lysis technique was also examined by comparison of an ammonium acetate precipitation of proteins and humic acids with a hexadecyltrimethylammonium bromide (CTAB) incubation and phenol:chloroform extraction. In this study, the incorporation of soil washing steps facilitated the desorption of contaminants from sediment surfaces and improved the recovery of DNA of amplifiable quality from both freshwater and marine sediments. CTAB contributed only slightly to the recovery of DNA of higher quality in the most contaminated sample from Sydney Harbour and was concomitant with a decrease in DNA yield in both sediment types. The incorporation of a soil washing step prior to the extraction of DNA from polluted environments may be important to solubilize and remove contaminants when high-quality DNA is required for subsequent analyses.     

An examination of the soil washing technique by its application to several soils

Summary The soil-washing technique is discussed in relation to its efficiency in removing fungal spores from soil samples and to its application for studying soil microhabitats. Detailed data on the application of the technique to a range of soils are given, and an attempt is made to evaluate the potentialities of the technique for studies on soil fungi.     

Cytology of peritoneal washings in gynecologic patients. Diagnostic criteria and pitfalls

A total of 226 peritoneal washing specimens obtained from gynecologic patients over a three-year period was reviewed. The diagnostic problems encountered were: differentiating reactive mesothelium from low-grade malignancies; distinguishing between benign ovarian tumors, ovarian tumors of borderline malignancy and low-grade ovarian malignancies; and potential false-positive diagnoses in endometriosis. Low-grade malignancies could be distinguished from reactive mesothelium by evaluating subtle cytologic criteria and by comparing the cells to those of the tumor on histologic section. The differentiation of low-grade epithelial malignancies from benign epithelial lesions caused difficulties that could only be resolved by evaluating the histologic material. Positive peritoneal washings increased the stage in 8 of 110 patients undergoing initial surgery for gynecologic malignancy. Two of 76 patients undergoing a second-look laparotomy had positive washings without histologic evidence of tumor. These ten patients did less well than did those with similar histology but negative washing cytology, despite receiving additional therapy because of the cytologic findings.     

Complexation of a Ligand with a Surfactant Micelle for Soil Heavy Metal Desorption

The ligands iodide (I^?) and thiocyanate (SCN^?), alone or in admixture, in combination with a non-ionic surfactant, Triton X-100, were evaluated as washing agents for heavy metal desorption from a contaminated soil. After seven successive washings, selective sequential extraction (SSE) was performed to determine the heavy metal content that remained associated with each geo-chemical fraction of the soil. The surfactant with 0.336 mol L^?1 of ligand I^? removed 75% Cd and 23% Cu, whereas the mobilization of Zn and Pb were not significant after 7 washings. At a concentration of 0.286 mol L^?1, the ligand SCN^? in the presence of surfactant removed 36% Cd, 44% Cu and 77% Zn. Among the washing agents, the combination of I^? and SCN^? produced the highest desorption efficiencies 95% Cd, 48% Cu, and 3.1% Pb, but not for Zn. The SCN^? ligand extracted the most Zn (77%). The SSE procedure indicated that the I^? removed metals from the exchangeable, carbonate and oxide fractions whereas SCN^? removed metals only from the exchangeable fraction. Both ligands, in the presence of surfactant, removed Cu from all fractions except the exchangeable sites, whereas only SCN^? plus surfactant removed Zn from all fractions. The ligand mixture plus surfactant mobilized only limited quantities of Pb from the oxide and residual fractions.     

Paranuclear blue inclusions of small cell carcinoma of the stomach: report of a case with cytologic presentation in peritoneal washings

BACKGROUND: Primary gastric small cell carcinoma is a rare but important entity. We describe a case that we diagnosed by peritoneal washing cytology. CASE: A 70-year-old male presented with upper abdominal discomfort and underwent endoscopic evaluation. Gastric endoscopy revealed a diffuse, infiltrating tumor from the body to the antrum. Total gastrectomy with lymph node dissection and intraoperative peritoneal washing cytology were carried out. Peritoneal washing cytology showed the presence of many undifferentiated malignant small cells with a necrotic background. The tumor cells were small and round, with naked, hyperchromatic nuclei and finely granular chromatin. Some tumor cells contained paranuclear blue inclusions (PBls) in the cytoplasm. The tumor cells were positive for neuron-specific enolase and synaptophysin on immunocyto-chemistry. Carcinoembryonic antigen, alpha-fetoprotein (AFP) and leukocyte common antigen were negative. Pathologic diagnosis after the operation was moderately to poorly differentiated adenocarcinoma and small cell carcinoma containing AFP-positive cells. CONCLUSION: The prognosis of primary gastric small cell carcinoma is usually poor. Our patient died of multiple liver metastases and peritonitis carcinomatosa 69 days after surgery. When a gastric small cell carcinoma is suspected in peritoneal washings, immunocytochemical demonstration of neuroendocrine differentiation is required to arrive at the final diagnosis.     

湛江红树林滩涂可培养真菌种群多样性分析

【目的】明确湛江地区红树林滩涂海洋真菌的种类及其分布,为海洋真菌的开发利用研究奠定基础。【方法】运用稀释平板法从湛江市高桥及特呈岛红树林滩涂不同的潮位带(低、中、高)和不同树种(白骨壤、桐花树、木榄、红海榄)采集淤泥样品550份,采用真菌形态学和ITS序列分析技术进行多样性研究【。结果】分离获得海洋真菌274株,共鉴定出19属39种真菌,以曲霉属Aspergillus、青霉属Penicillium和木霉属Trichoderma真菌分离频率高,为湛江红树林滩涂优势真菌种类,尤其在中潮位带真菌种类最多。此外,分离获得真菌Talaromyces helicus,为中国新记录种。【结论】湛江红树林滩涂海洋真菌的种类十分丰富,具有潜在的开发和利用前景。