Relating algal-derived extracellular and intracellular dissolved organic nitrogen with nitrogenous disinfection by-product formation.

The dissolved organic nitrogen (DON) pool from algal-derived extracellular and intracellular organic matter (EOM and IOM) comprises proteins, peptides, free amino acids and carbohydrates, of which, proteins can contribute up to 100% of the DON. Previous reports of algal-derived DON character have focused on bulk properties including concentration, molecular weight and hydrophobicity. However, these can be similar between algal species and between the EOM and IOM even when the inherent molecular structures vary. A focus on bulk character presents challenges to the research on algal-derived nitrogenous-disinfection by-product (N-DBP) formation as N-DBP formation is sensitive to the changes in molecular structure. Hence, the main aim of this study was to characterize algal EOM and IOM-derived DON, specifically proteinaceous-DON, using a combination of bulk and molecular characterization techniques to enable a more detailed exploration of the relationship between the character of algal-derived proteins and the N-DBP formation potential. DON from the EOM and IOM of four commonly found algae and cyanobacteria in natural waters were evaluated, namely Chlorella vulgaris, Microcystis aeruginosa, Dolichospermum circinale, and Cylindrospermopsis raciborskii. It was observed that 77-96% of total DON in all EOM and IOM samples was of proteinaceous origin. In the proteins, DON concentrations were highest in the high molecular weight fraction of IOM-derived bulk proteins (0.13-0.75 mg N L-1) and low to medium molecular weight fraction of EOM-derived bulk proteins (0.15-0.63 mg N L-1) in all species. Similar observations were also made via sodium dodecyl sulphate polyacrylamide gel electrophoresis and liquid chromatography-high resolution mass spectrometry. Solid-state 15N nuclear magnetic resonance (NMR) spectroscopy of the EOM and IOM revealed the existence of common aliphatic and heterocyclic N-groups in all samples, including a dominant 2° amide peak. Species dependent variability was also observed in the spectra, particularly in the EOM; e.g. nitro signals were found only in the Cylindrospermopsis raciborskii EOM. Dichloroacetonitrile (DCAN) and N-nitrosamine concentrations from the EOM of the species evaluated in this study were lower than the guideline limits set by regulatory agencies. It is proposed that the dominant 2° amide in all samples decreased N-DBP formation upon chlorination. For chloramination, the presence of nitro groups and aliphatic and heterocyclic N-DBP precursors could cause variable N-nitrosamine formation. Compared to non-algal impacted waters, algae-laden waters are characterised by low organic carbon: organic nitrogen ratios of ∼7-14 and elevated DON and protein concentrations. Hence, relying only on bulk characterization increases the perceived risk of N-DBP formation from algae-laden waters.

Microalgae harvesting using flocculation and dissolved air flotation: Selecting the right vessel for lab-scale experiments.

Flocculation combined with dissolved air flotation (DAF) is a promising technology for harvesting microalgae; therefore, optimisation of flocculant-DAF operating conditions are frequently explored in laboratory experiments. DAF systems have jars of differing volumes, height to diameter ratios, shapes and materials used to manufacture the jars; thus, the harvesting efficiency (η) may differ between these jars. The aim was to systematically compare η between different types of benchtop DAF jars. Evaluation of 30 different types of DAF jars revealed that η was not influenced by the volume of the jars, but was impacted by the height to diameter ratio, with optimal η at a ratio ranging between 1.6 and 2.05. There was no difference in η between cylindrical and cuboid jars, but jars made of hydrophobic (polypropylene) plastic resulted in a lower η. Overall, these results are useful to guide the design of lab-scale DAF microalgae harvesting experiments.

Understanding variability in algal solid-liquid separation process outcomes by manipulating extracellular protein-carbohydrate interactions.

Coagulation-flocculation followed by sedimentation or dissolved air flotation (DAF) are processes routinely used for separating microalgae from water; however, during algae separation then can exhibit inconsistent separation, high coagulant demand, and high operating cost. To circumvent these problems, previous studies reported the development of a novel DAF process in which bubbles were modified instead of particles. While this process was shown to be sustainable and inexpensive, the problem of inconsistent algal separation across species remained. Recent research has suggested that this could be due to the varying concentration and character of algal-derived proteins and carbohydrates within the extracellular organic matter (EOM) and their associated interactions. This hypothesis is tested in the current study using the novel modified-bubble DAF process, which has been highly susceptible to EOM protein and carbohydrate concentrations and character. Biomolecular additives (commercially available proteins and carbohydrates, and algal-extracted proteins) of widely differing molecular weight (MW) and charge were dosed in varying proportions into samples containing either Chlorella vulgaris CS-42/7, Microcystis aeruginosa CS-564/01, or Microcystis aeruginosa CS-555/1 after removing the intrinsic EOM. These cell-rich suspensions were then subject to flotation using cationic bubbles modified with poly(diallyldimethylammonium chloride) (PDADMAC). When additives were dosed independently, separation increased from <5% to up to 62%. The maximum separation was obtained when the dose was double the respective biopolymer concentration measured in the intrinsic EOM for the equivalent species, and, in the case of protein additives, when MW and charge were >50 kDa, and >0.5 meq·g-1, respectively, irrespective of the species tested. When evaluating steric- and charge-based protein-carbohydrate interactions on cell separation by simultaneously dosing high MW and high charge protein- and carbohydrate-additives, enhanced separation of up to 79% was achieved. It is suggested that enhanced cell separation is achieved due to proteins and carbohydrates bridging with cells and forming protein-carbohydrate-cell suprastructures in the presence of a flocculant, e.g. PDADMAC, and this only occurs when the intrinsic EOM comprises proteins and carbohydrates that have high MW (>25 kDa) and charge (>0.2 meq·g-1), and interactions with each other and with the cell surface.

Formation of algal-derived nitrogenous disinfection by-products during chlorination and chloramination.

Algal cells and algal organic matter (AOM) are a source of high dissolved organic carbon (DOC) and nitrogen (DON) concentrations. This poses a possible health risk due to their potential to form disinfection by-products (DBPs), some of which may be of health concern, after disinfection. While several studies have focussed on the formation of carbonaceous DBPs from AOM, only a few studies have focussed on the formation of nitrogen containing N-DBPs from AOM. Hence, the main aim of this study was to thoroughly investigate the N-DBP formation potential of the AOM from a species of cyanobacteria commonly found in natural waters, Microcystis aeruginosa. Three haloacetonitriles, two halonitromethanes, two haloacetamides, and eight N-nitrosamines were analysed by gas chromatography-mass spectrometry after chlorination and chloramination of the extracted AOM. To provide further insight into the influence of changing DON character on N-DBP formation potential, the AOM from three other species, Chlorella vulgaris, Dolichospermum circinale and Cylindrospermopsis raciborskii, were also tested. Dichloroacetonitrile (DCAN) was the DBP formed in the highest concentrations for both chlorination and chloramination of bulk AOM from all the species. Furthermore, during chlorination and chloramination, the high molecular weight fraction (>1 kDa) of AOM from M. aeruginosa had a greater DCAN formation potential (normalised to DOC or DON) than the AOM in the low molecular weight fraction (<1 kDa) of M. aeruginosa, regardless of growth stage. N-Nitrosamine formation from the bulk AOM of all species occurred only after chloramination. The molar concentration of N-nitrosodimethylamine (NDMA) was lower than the other N-nitrosamines detected. However, NDMA formation increased with culture age for all four species, in contrast to most other N-nitrosamines whose formation remained consistent or decreased with culture age. Overall, algal growth could result in elevated concentrations of N-DBPs due to the increasing concentrations of high molecular weight algal DON in the AOM. It is suggested that the AOM comprises precursors containing long C-chain amine (R1-NH-R2) or cyclic N-containing amine structures. Comparisons to previously measured N-DBP concentrations in drinking water suggest that the AOM from the algae and cyanobacteria examined in this study are not likely to be a major source of precursors for either DCAN or NDMA in real waters. However, AOM may present a major precursor source for other N-nitrosamines.

Detailed algal extracellular carbohydrate-protein characterisation lends insight into algal solid-liquid separation process outcomes.

The effectiveness of algal solid-liquid separation processes has been impacted by the strong influence of algal extracellular organic matter (EOM), where the composition of proteins and carbohydrates and their associated interactions have been implicated. However, despite this, no studies have analysed the detailed protein and carbohydrate composition in EOM in relation to their impacts on separation. Hence, the aim of this study was to explore the relationship between the variety of carbohydrates and proteins present in the EOM of select algal and cyanobacterial samples and the associated separation performance to better understand the influence of specific biopolymers. The protein and carbohydrate composition of the EOM of three species - Microcystis aeruginosa CS-555/1, Chlorella vulgaris CS-42/7 and Microcystis aeruginosa CS-564/01, previously observed to result in variable treatment performance were investigated. The carbohydrates were analysed via high-performance anion-exchange chromatography (HPAEC) with pulsed amperometric detection (PAD) while the proteins were analysed using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) combined with liquid chromatography-mass spectrometry (LC-MS). Ten unique monosaccharides were identified; of these, the greatest proportion of charged uronic acid carbohydrates were present in the EOM of M. aeruginosa CS-564/01. The protein profiling revealed that M. aeruginosa CS-564/01 had a greater proportion and concentration of proteins >75 kDa when compared to M. aeruginosa CS-555/1 or C. vulgaris CS-42/7. It was determined that three serine- and two threonine-based proteins, detected in greater concentrations in M. aeruginosa CS-564/01 than CS-555/1, could covalently interact with carbohydrates (OHenderson et al., 2010a, 2010b-linked glycosylation). These proteins have the ability to form numerous localised networks with carbohydrates and cells in the presence of coagulant molecules, thereby providing a good hypothesis to explain the excellent treatment performance observed for M. aeruginosa CS-564/01 previously. It is proposed that the uronic acids in M. aeruginosa CS-564/01 could interact with proteins via glycosylation, explaining why the coagulant demand for this strain remained low despite the high charged carbohydrate concentration. Overall, it is proposed that process performance could be impacted by: (a) physicochemical characteristics and (b) carbohydrate-protein interactions.

An evaluation of measurement techniques for algal-derived organic nitrogen.

Algal-derived organic matter (AOM) from algal blooms in water supply systems contains dissolved organic nitrogen (DON) and dissolved organic carbon (DOC) among other constituents. The DON and DOC are disinfection by-product (DBP) precursor compounds, and must be well characterised to facilitate effective removal, thus minimising DBP formation during disinfection. While DOC character has been studied extensively, DON analysis suffers from inaccuracies due to sample pre-treatment and instrument sensitivities. A liquid chromatography method that combines size exclusion chromatography with highly sensitive organic carbon and nitrogen detectors (LC-OCND) has been widely adopted for DOC analysis; however, its potential for application for DON charactersation has been suggested as a viable alternative to existing DON characterisation techniquesnot been assessed despite its potential. Hence, the aim was to compare the effectiveness of conventional total dissolved N-dissolved inorganic N (TN-DIN), and LC-OCND methods for analysing DON in AOM. A suite of N-containing model compounds representative of DON and AOM extracted from Chlorella vulgaris CS-42/7 and Microcystis aeruginosa CS-555/1 were used to evaluate the techniques. The DON of both model compounds and AOM was first analysed using the conventional method and, then, via LC-OCND. It was observed that LC-OCND had a better precision for DON when TN contained more DIN. LC-OCND provided direct quantitative measurements for bulk and fractionated DON and DIN, with little interference caused by DIN. Additionally, LC-OCND provided information on MW distribution and protein content of the AOM. For example, LC-OCND results showed that M. aeruginosa AOM contained more HMW material than C. vulgaris AOM. However, as LC-OCND uses UV oxidation, it could not completely oxidise complex aromatic structures, and thus had a lower recovery for HMW model compounds and algal DON in comparison to the conventional method that used high temperature catalytic oxidation. Overall, it is advised that a combination of LC-OCND and TN analysis be used to provide a more detailed characterisation of N-containing AOM and other similar HMW aquatic NOM samples.

Multiple cutaneous swellings in an immunocompetent host--cryptococcosis overlooked.

Recently, there has been an increase in the incidence of cryptococcosis even among immunocompetent patients. We present a case of multiple subcutaneous swellings later developing into a non-healing ulcer caused by Cryptococcus in a 60 year-old immunocompetent male along with a brief review of clinical presentations of cutaneous cryptococcosis in immunocompetent and immunocompromised individuals. A high index of suspicion is desired in view of possible dissemination and variable non-specific clinical presentations.

A comparative evaluation of antimicrobial efficacy and flow properties for Epiphany, Guttaflow and AH-Plus sealer.

AIM: To test the antimicrobial efficacy and flow properties of Guttaflow, Epiphany sealer and AH-Plus sealer. METHODOLOGY: With the use of Enterococcus faecalis ATCC 29212 as a test organism, both the agar diffusion test (ADT) and direct contact test (DCT) were performed. For DCT, sealers were mixed and placed over the bottom of sterile screw-capped test tubes. A 50 µL bacterial suspension was placed on the tested material samples. Bacteria were allowed to directly come in contact with the sealers for 1 h at 37 °C in one group and for 24 h in the other group. The suspensions were then diluted and inoculated over blood agar plates, and bacterial colony counts were determined with the use of a digital colony counter. The data in both 1- and 24-h groups were individually analysed using repeated measures ANOVA. Kruskal Wallis tests were further used to obtain comparison between 1- and 24-h results for all three sealers. In the flow assay, the sealers were placed between two glass slides, and a weight of 500 g was placed on the top of the glass. The diameters of the formed discs were recorded. RESULTS: For both the ADT and DCT tests, Epiphany and AH-Plus sealer reduced the bacterial counts significantly (P = 0.000). Epiphany produced a greater reduction in bacterial counts when compared to AH-Plus in both the tests (P = 0.000). Guttaflow paste failed to show any antibacterial activity in both ADT & DCT. According to the flow test, all root canal sealers flowed; Epiphany sealer had the maximum flow under the given conditions, followed by AH-Plus sealer and Guttaflow paste. CONCLUSIONS: Antimicrobial activity of the sealers was greatest for Epiphany followed by AH-Plus sealer and Guttaflow. Epiphany sealer had the maximum flow followed by AH-Plus sealer and Guttaflow.

Studies on Alternaria sesami pathogenic to sesame.

Influence of pH on the growth of Alternaria sesami, its nutritional requirements and its ability to produce phytotoxic and antibacterial metabolites were tested. The isolate was cultured on Czapek-Dox broth and the culture filtrates were screened for phytotoxicity against seeds and seedlings of sesame. Chloroform extracts of the fungus exhibited antibacterial activity. Analysis of the culture filtrates for identifying toxins using chromatographic techniques revealed the presence of tenuazonic acid.

International medical students' career choice, attitudes toward psychiatry, and emigration to the United States : examples from India and zimbabwe.

The selection residents who are international medical graduates can be facilitated by knowledge of the context of their training and experience in psychiatry; the career opportunities in the country from which they come; and their attitudes toward psychiatry, medicine, and the United States. This study examines some of these variables as well as the emigration plans of 209 Indian and 46 Zimbabwe medical students in their next to last year of medical school. Both groups tended to hold positive attitudes toward psychiatry and had good experiences with the field. Fifty-six percent of the Indian students indicated that they had at least thought about emigrating to the United States for postgraduate training, and 22% had definite plans. Only 25% of the Zimbabwe students had thought about U.S. emigration, and a mere 2% had plans. Indian, as well as other IMGs, will continue to be a presence in U.S. residency programs, and those who have a preresidency stay in the United States may come to know and appreciate psychiatry more as they become more familiar with the field and with American culture.

Countertransference. Its continued importance in psychiatric education.

Psychotherapy is likely to play a minor or nonexistent role in the future general psychiatrist's training and practice. However, the component skills of recognition and management of the countertransference will remain important. Because psychotherapy training and supervision have been the venues for countertransference learning, the field is in danger of losing its teaching laboratory and hence losing these skills. The authors examine the concept of countertransference and discuss its importance in four increasingly significant areas: managed care, psychopharmacological treatment, emergency intervention, and the management of professional boundaries regarding sexual misconduct. Methods are discussed for enhancing residents' countertransference skills through supervision, training groups, and the resident's personal psychotherapy.

Some medicinal orchids of southern India.

Ethnobotanical Studies were carried out in some parts of Ahdhra Pradesh and Tamil Nadu to identify orchids reported to have medicinal properties and to study their botanical distribution. This has resulted in the recording of 9 orchids used by the local population.

Effects of benzene metabolite treatment on granulocytic differentiation and DNA adduct formation in HL-60 cells.

Reactive metabolites of benzene (BZ) play important roles in BZ-induced hematotoxicity. Although reactive metabolites of BZ covalently bind to DNA, the significance of DNA adduct formation in the mechanism of BZ toxicity is not clear. These studies investigated the covalent binding of the BZ metabolites hydroquinone(HQ) and 1,2,4-benzenetriol(BT) using the DNA [32P]postlabeling method and explored the potential relationship between DNA adduct formation and cell differentiation in human promyelocytic leukemia (HL-60) cells, a model system for studying hematopoiesis. Maturation of HL-60 cells to granulocytes, as assessed by light and electron microscopy, was significantly inhibited in cells that were pretreated with HQ or BT prior to inducing differentiation with retinoic acid (RA). The capacity of RA-induced cells to phagocytose sheep red blood cells (RBC) and to reduce nitroblue tetrazolium (NBT), two functional parameters characteristic of mature, differentiated neutrophils, was also inhibited in cells pretreated with HQ or BT. These BZ metabolite treatments induced DNA adduct formation in HQ- but not in BT-treated cells. These results indicate that whereas HQ and BT each block granulocytic differentiation in HL-60 cells, DNA adducts were observed only following HQ treatment. Thus DNA adduct formation may be important in HQ but not in BT toxicity.

Alpha blocker prazosin for the treatment of benign prostatic hypertrophy (BPH).

In this study medical treatment with alpha blocker-prazosin is compared with transurethral resection of prostate (TURP) in 62 patients suffering from benign enlargement of prostate with a gland size of less than 20 gms. After thorough interrogation patients were offered either TURP or prazosin therapy. Symptom scoring, residual volume of urine and urinary flow rates were estimated in both the groups before and 3 months after the therapy. 23.5% patients in prazosin group while 90% of patients in TURP group had significant improvement. This distinctly brings out the superiority of TURP for benign enlargement of prostate.

Oxidative modifications produced in HL-60 cells on exposure to benzene metabolites.

We have studied the effects of the benzene metabolites hydroquinone, p-benzoquinone or 1,2,4-benzenetriol on cytotoxicity, active oxygen formation, hydrogen peroxide (i.e. hydroperoxide) production and nitric oxide formation in HL-60 cells. We also examined the effects of these compounds on antioxidant enzymes and intracellular antioxidants in these cells. The cytotoxicity of benzene metabolites to HL-60 cells was found to be of the order of p-benzoquinone>hydroquinone>benzenetriol. No appreciable changes in the basal levels of either superoxide anion production or nitric oxide formation were observed following exposures to the benzene metabolites, but significant increases in superoxide were seen on stimulation with TPA for each metabolite, whereas hydroquinone and p-benzoquinone, but not 1,2,4-benzenetriol, increased nitric oxide production under these conditions. Following exposure to the benzene metabolites, HL-60 cells showed significant rises in hydrogen peroxide formation compared to controls. The study of antioxidant enzymes and intracellular antioxidants suggested that the benzene metabolites inhibit or reduce the levels of different antioxidant mechanisms and, thereby, cause the accumulation of free radicals in these cells predisposing them for oxidative damage.

Anterior urethral valve in an adolescent boy.

A 14 year old boy with a relatively uncommon anterior urethral valve is described herein.

Distinct actions of benzene and its metabolites on nitric oxide production by bone marrow leukocytes.

Benzene is a widely used industrial solvent known to cause bone marrow depression. This is associated with increased production of reactive oxygen metabolites and nitric oxide by bone marrow phagocytes, which have been implicated in hematotoxicity. Benzene metabolism to phenolic intermediates appears to be an important factor in bone marrow toxicity. In the present studies, we compared the effects of benzene and several of its metabolites on nitric oxide production by murine bone marrow leukocytes. Bone marrow cells readily produced nitric oxide in response to the inflammatory mediators lipopolysaccharide (LPS) and interferon-gamma (IFN-gamma). Treatment of mice with benzene (800 mg/kg), or its metabolites hydroquinone (100 mg/kg), 1,2,4-benzenetriol (25 mg/kg), or p-benzoquinone (2 mg/kg), at doses that impair hematopoiesis, sensitized bone marrow leukocytes to produce increased amounts of nitric oxide in response to LPS and IFN-gamma. Granulocyte-macrophage colony-stimulating factor (GM-CSF) and macrophage colony-stimulating factor (M-CSF) augmented bone marrow leukocyte production of nitric oxide induced by inflammatory mediators. Benzene, as well as its metabolites, markedly increased the sensitivity of the cells to both GM-CSF and M-CSF. Cells from hydroquinone- or 1,2,4-benzenetriol-treated mice were significantly more responsive to the inflammatory cytokines and growth factors than cells isolated from benzene- or p-benzoquinone-treated mice, suggesting that the phenolic metabolites of benzene are important biological reactive intermediates. Because nitric oxide suppresses cell growth and can be metabolized to mutagens and carcinogens, the ability of benzene and its metabolites to modulates its production in the bone marrow may be important in their mechanism of action.

Perspectives on screening and interviewing international medical graduates for psychiatric residency training programs.

The authors have found that international medical graduates (IMGs) constitute a valuable pool of applicants for residency training. The wide variation in their medical educations, prior careers, and cultural backgrounds requires a special approach to screening and interviewing IMGs. The authors share techniques and viewpoints developed in their program to screen applicants about their autobiographical statements, medical credentials, reference letters, and visa and immigration statuses. Interviewing guidelines that are sensitive to the applicant's career phase and that assess the person's interest in psychiatry, communication skills, character, acculturation, family context, and suitability for the specific residency program are presented.