Microplastics pollution in the rivers of a metropolitan city and its estimated dependency on surrounding developed land.

The spatial distribution and abundance of suspected microplastics (SMPs) in the surface water of a metropolitan city, as represented by four Hong Kong rivers, was studied during the dry season. Shing Mun River (SM), Lam Tsuen River (LT), and Tuen Mun River (TM) are located in urbanized areas, and SM and TM are tidal rivers. The fourth river, Silver River (SR) is situated in a rural area. TM had a significantly higher SMP abundance (53.80 ± 20.67 n/L) than the other rivers. The SMP abundance increased from upstream to downstream in non-tidal rivers (LT and SR), but not in tidal rivers (TM and SM), probably due to the tidal influence and a more homogeneous urban development along the tidal rivers. Inter-site differences in the SMP abundance were strongly correlated with the built area ratio (defined as the percentage of surrounding developed land area), human activities, and the nature of the river. About half (48.72 %) of the SMPs were <250 µm. Fibers and fragments were most abundant (>98 %), with most of them being transparent (58.54 %), black (14.68 %), or blue (12.12 %). Polyethylene terephthalate (26.96 %) and polyethylene (20.70 %) were the most common polymers. However, the MP abundance could be overestimated due to the presence of natural fibers. By contrast, an underestimation of the MP abundance could result from a smaller volume of water samples collected, due to a low filtration efficiency caused by high organic content and particle concentrations in the water. A more effective solid waste management strategy and upgrading of the sewage treatment facilities for removing microplastics are recommended to ameliorate the microplastic pollution in local rivers.

Microplastic ingestion reduces energy intake in the clam Atactodea striata.

The effects of microplastic concentrations (10itemsl-1 and 1000itemsl-1) on the physiological responses of Atactodea striata (clearance rate, absorption efficiency, respiration rate) were investigated. The fates of ingested microplastics and the efficiency of depuration in removing ingested microplastics were also studied. A. striata ingested microplastics and the clearance rate was reduced at high concentration of microplastics. Since the respiration rate and absorption efficiency remained unchanged in exposed A. striata, reduction in the clearance rate would reduce the energy intake. Ingestion and retention of microplastics in the body were further limited by the production of pseudofaeces and faeces, and depuration in clean water, resulting in a very small amount of microplastics stored in the body of the clam.

64-slice CT angiography for the detection of functionally significant coronary stenoses: comparison with stress myocardial perfusion imaging.

OBJECTIVE: To evaluate the accuracy of 64-slice CT angiography (CTA) compared with single photon emission CT (SPECT) myocardial perfusion imaging (MPI), which served as the reference standard, for the detection of functionally significant coronary artery disease (CAD). METHODS: 141 consecutive patients (60 ± 10 years, 101 men) were investigated with 64-slice CTA and SPECT MPI; a subset of 35 patients had additional invasive coronary angiography (ICA). The data from CTA and ICA were compared with those from MPI for both cut-offs of ≥ 50% and ≥ 70% stenosis, respectively. RESULTS: The sensitivity, specificity, positive and negative predictive values, and accuracy of CTA, using a cut-off of ≥ 50% for significant stenosis, in detecting inducible perfusion defects on MPI were 96% [95% confidence interval (CI) 88-100%], 61% (95% CI 52-70%), 37% (95% CI 23-49%), 99% (95% CI 97-100%) and 68%, respectively, in patient-based analysis and 97% (95% CI 91-100%), 86% (95% CI 83-89%), 33% (95% CI 24-42%), 100% (95% CI 99-100%) and 87%, respectively, in vessel-based analysis. Applying a cut-off of ≥ 70% for significant stenosis, CTA yielded the following sensitivity, specificity, positive and negative predictive values, and accuracy for the detection of inducible MPI defects: by patient, 65% (95% CI 46-84%), 95% (95% CI 91-99%), 74% (95% CI 50-92%), 92% (95% CI 87-97%) and 89%, respectively; by vessel, 58% (95% CI 42-74%), 97% (95% CI 95-99%), 62% (95% CI 45-79%), 97% (95% CI 95-99%) and 95%, respectively. CONCLUSION: 64-slice CTA is a reliable tool to exclude functionally significant CAD when using a cut-off of ≥ 50% diameter stenosis. By contrast, a cut-off of ≥ 70% diameter narrowing is a strong predictor of ischaemia.

Identification of F. o. cucumerinum and F. o. luffae by RAPD-generated DNA probes.

AIMS: To create a fast, sensitive and specific method for identifying Fusarium oxysporum f. sp. cucumerinum and F. o. luffae. METHODS AND RESULTS: Specific DNA bands were selected as probes from RAPD profiles of 13 formae speciales of F. oxysporum. The forma specialis-specific probe OPC18300c and OPC18520f could be used to identify F. o. cucumerinum and F. o. luffae by RAPD-PCR followed dot blot hybridization, respectively. CONCLUSIONS: A specific method for identifying F. o. cucumerinum and F. o. luffae was achieved. SIGNIFICANCE AND IMPACT OF THE STUDY: F. oxysporum formae speciales identification with a DNA probe can be relatively rapid and provides a method to identify the pathogen without host inoculation tests.

Genetic polymorphism and Parkinson's disease in Taiwan: study of debrisoquine 4-hydroxylase (CYP2D6).

Debrisoquine 4-hydroxylase (CYP2D6) is one of the cytochrome P450 enzyme families that catalyze the breakdown of a variety of exogenous and endogenous compounds. Previous reports have suggested that genetic polymorphisms of debrisoquine 4-hydroxylase are associated with susceptibility to Parkinson's disease (PD) in Caucasians. To determine if CYP2D6 also confers susceptibility to PD in Chinese patients, we carried out a study of genetic association using three polymorphic markers of the CYP2D6 gene, 188C/T, 1934G/A (mutant B), and 4268G/C. No differences of allele or genotype frequencies of these three polymorphisms were detected upon comparison of primary PD patients (n=53) with normal controls (n=94). The 1934A allele (mutant B), which accounts for the majority of poor metabolizers in Caucasians, is extremely rare in Chinese. Our data do not support the suggestion that the CYP2D6 gene is related to PD susceptibility in Chinese.

Associations among PH and SH3 domain-containing proteins and Rho-type GTPases in Yeast.

The src homology region 3 (SH3) domain-bearing protein Bem1p and the Rho-type GTPase Cdc42p are important for bud emergence in Saccharomyces cervisiae. Here, we present evidence that through its second SH3 domain, Bem1p binds to the structurally and functionally similar proteins Boi1p and Boi2p, each of which contain an SH3 and pleckstrin homology (PH) domain. Deletion of BOI1 and BO12 together leads to impaired morphogenesis and poor ability. A PH domain-bearing segment of Boi1p that lacks the Bem1p-binding site is necessary and sufficient for function. This segment of Boi1p displays a two-hybrid interaction with Cdc42p, suggesting that Boi1p either binds directly to or is part of a larger complex that contains Cdc42p. Consistent with these possibilities, overexpression of Boi1p inhibits bud emergence, but this inhibition is counteracted by cooverexpression of Cdc42p. Increased expression of the Rho-type GTPase Rho3p, which is implicated in bud growth defects of boil boi2 mutants, suggesting that Boi1p and Boi2p may also play roles in the activation or function of Rho3p. These findings provide an example of a tight coupling in function between PH domain-bearing proteins and both Rho-type GTPases and SH3 domain-containing proteins, and they raise the possibility that Boi1p and Boi2 play a role in linking the actions of Cdc42p and Rho3p.

Pentatrichomonas hominis: purine salvage pathway.

1. Pentatrichomonas hominis was found incapable of de novo synthesis of purines. 2. Pentatrichomonas hominis can salvage adenine, guanine, hypoxanthine, adenosine, guanosine and inosine, but not xanthine for the synthesis of nucleotides. 3. HPLC tracing of radiolabelled purines or purine nucleosides revealed that adenine, adenosine and hypoxanthine are incorporated into adenine nucleotides and IMP through a similar channel while guanine and guanosine are salvaged into guanine nucleotides via another route. There appears to be no direct interconversion between adenine and guanine nucleotides. Interconversion between AMP and IMP was observed. 4. Assays of purine salvage enzymes revealed that P. hominis possess adenosine kinase; adenosine, guanosine and inosine phosphotransferases; adenosine, guanosine and inosine phosphorylases and AMP deaminase.

Persistence of hepatitis B viral antigens in Culex quinquefasciatus.

Culex quinquefasciatus mosquitoes were fed on or inoculated with blood or serum positive for hepatitis B viral antigens and pools of mosquitoes were tested by radioimmunoassay daily for 3 weeks after exposure to detect the viral antigens. Hepatitis B surface antigen (HBsAg) was detectable up to 3 weeks, while hepatitis B e antigen (HBeAg) persisted only for 3 days in mosquitoes after feeding on hepatitis B viral antigens-positive blood. Mosquitoes inoculated with serum were HBsAg-positive for 3 weeks and HBeAg positive for 4 days after inoculation. These results suggest that biological multiplication of hepatitis B virus did not occur in these mosquitoes. The possibility of mechanical transmission of hepatitis B antigens by mosquitoes is discussed.

Purine salvage in Entamoeba histolytica.

Pulse-labeling of the nucleotide pool in Entamoeba histolytica with radioactive precursors, and subsequent high performance liquid chromatographic (HPLC) analysis of the radiolabeled nucleotides, indicate that E. histolytica is incapable of de novo synthesis of purine nucleotides. Hypoxanthine, inosine and xanthine could not be converted to nucleotides in E. histolytica, which suggests the absence of interconversion between adenine nucleotides and guanine nucleotides through formation of IMP. Adenosine was actively incorporated into nucleotides at an initial rate of 130 pmoles per minute per 10(6) trophozoites. Adenine, guanosine and guanine were also incorporated at much lower rates. The rate of adenine incorporation was enhanced by the presence of guanosine; the rate of guanine incorporation was significantly increased by adenosine. These stimulatory effects suggest that the ribose moiety of adenosine or guanosine can be transferred to another purine base to form a new nucleoside, and that the purine nucleosides are the immediate precursors of E. histolytica nucleotides. HPLC results showed that the radiolabel in adenine was exclusively incorporated into adenine nucleotides and that guanine was found only among guanine nucleotides, whereas the radioactivity associated with the ribose moiety of adenosine or guanosine was distributed among both adenine and guanine nucleotides.

Purification and properties of NADP-linked, alcohol dehydrogenase from Entamoeba histolytica.

An NADP-linked, alcohol dehydrogenase from Entamoeba histolytica was purified to apparent homogeneity by Blue Sepharose affinity chromatography. Molecular weights of 130,000 and 30,000 were estimated by gel filtration and by sodium dodecyl sulfate polyacrylamide gel electrophoresis, respectively, suggesting that the enzyme is a tetramer. The enzyme exhibited more than 20-fold selectivity for NADP(H) over NAD(H). Although the purified enzyme acts on both primary and secondary alcohols, higher activity was found with secondary alcohols.

Glucose dissimilation in Entamoeba histolytica.

The endproducts of glucose dissimilation in Entamoeba histolytica were found to be carbon dioxide, acetate and ethanol. This organism possesses the same Embden-Meyerhof intermediates as found in most investigated organisms. In its glucose-to-pyruvate pathway several unusual features were demonstrated. The pyruvate-to-acetate and pyruvate-to-ethanol pathways are also unique as compared with that of other organisms. The distinctive properties of the metabolic enzymes provide the basis for the development of new chemotherapeutic agents against this parasitic protozoa.

[Trichomonas hominis: isolation and axenic cultivation (author's transl)].

Recently we have isolated Trichomonas hominis from diarrheic stools of a patient and established it in an axenic culture medium. The procedures are as follows: Diarrheic stool containing numerous trophozoites was first inoculated into the TYM (Trypticase Yeast extract Maltose) medium of Diamond (1975) to establish a polyxenic culture. Antibiotics, containing penicillin (1000 U/ml), streptomycin (1000 micrograms/ml), and cephalosporin (20 micrograms/ml) were added to prevent overgrowth of bacteria and fungi. After several passages, a specially-designed culture-tube was employed to separate T. hominis from the contaminants. The isolated T. hominis was then introduced into the modified TYI-S-33 (Trypticase Yeast extract Iron-Serum-33) medium of Diamond (1978). The organism established itself readily to this axenic culture medium. Sterility tests employing fluid thioglycollate, nutrient broth, and blood agar plate gave negative results indicating the absence of contaminants. The axenic culture of T. hominis provides us with a source of pure flagellates for biological, biochemical, and immunological studies.

Serum gangliosides in cerebral astrocytoma.

The serum concentration and composition of gangliosides were examined in 80 humans including 10 normal subjects. A significant increase was found in the total gangliosides of serum in 7 patients with cerebral astrocytomas. There was also an increased percentage of serum gangliosides with simpler structure, particularly GM3. The serum of patients with other intracranial tumors, including pituitary adenomas, ependymoma, teratoma, and metastases, did not show an increase in total ganglioside; however the pattern of simplification was found in these and in a few patients with extracranial tumors as well. The findings suggest that astrocytoma tumors shed sialoglycolipids into the circulation, and their assay may be useful in monitoring oncological therapy.

On the pathogenicity of Entamoeba histolytica. Part I: The role of bacterial associate on the modification of virulence of E. histolytica.

Of the 3 strains of Escherichia coli used, only Milner A strain was found capable of modifying the virulence of Entamoeba histolytica. None out of twenty-four hamsters inoculated with either 5 X 10(5) of axenically-cultured E. histolytica of NIH: 200 strain, or 1 X 10(7) of Esch. coli (A, B or C strains), was found to have amebic liver abscess. Whereas one out of six hamsters inoculated with the same number of amebae preincubated for 12 hrs with Esch. coli of Milner A strain was found to have abscess. The role of bacterial associate seems to be nothing but provides a more suitable environment for amebae, thus enable them to survive longer and endow them more time to adapt themselves to the given new environment. From liver abscess E. histolytica was recovered and successfully reaxenized. These amebae were capable of producing liver abscess, therefore the virulence seemed to be inheritable.

Riboflavin requirement for the cultivation of axenic Entamoeba histolytica.

Riboflavin was found to be essential for the cultivation of axenic Entamoeba histolytica. This is the first demonstration of a flavin requirement by the organism. Panmede, the principal source of flavins in the axenic medium, was treated with activated carbon to remove flavins. Medium made with this flavin-deficient Panmede, and supplemented with ribonucleic acid failed to support the multiplication of amebae in serial subculture, but did so when riboflavin was added. The concentration of riboflavin required to achieve maximal growth was about 1.3 microgram per ml medium. Studies on riboflavin uptake revealed that amebae lack a high-affinity transport system for this vitamin. The rate of riboflavin uptake was equivalent to the rate of pinocytotic uptake of fluid as previously determined.

Pyruvate-to-ethanol pathway in Entamoeba histolytica.

The pyruvate-to-ethanol pathway in Entamoeba histolytica is unusual when compared with most investigated organisms. Pyruvate decarboxylase (EC 4.1.1.1), a key enzyme for ethanol production, is not found. Pyruvate is converted into acetyl-CoA and CO2 by the enzyme pyruvate synthase (EC 1.2.7.1), which has been demonstrated previously in this parasitic amoeba. Acetyl-CoA is reduced to acetaldehyde and CoA by the enzyme aldehyde dehydrogenase (acylating) (EC 1.2.1.10) at an enzyme activity of 9 units per g of fresh cells with NADH as a reductant. Acetaldehyde is further reduced by either a previously identified NADP+-linked alcohol dehydrogenase or by a newly found NAD+-linked alcohol dehydrogenase at an enzyme activity of 136 units per g of fresh cells. Ethanol is identified as the product of soluble enzymes of amoeba acting on pyruvate or acetyl-CoA. This result is confirmed by radioactive isotopic, spectrophotometric and gas-chromatographic methods.

An energy-conserving pyruvate-to-acetate pathway in Entamoeba histolytica. Pyruvate synthase and a new acetate thiokinase.

Under anaerobic conditions, cells of Entamoeba histolytica grown with bacteria produce H2 and acetate while cells grown axenically produce neither. Aerobically, acetate is produced and O2 is consumed by amebae from either type of cells. Centrifuged extracts, 2.4 x 106 x g x min, from both types of cells contain pyruvate synthase (EC 1.2.7.1) and an acetate thiokinase which, together, form a system capable of converting pyruvate to acetate. Pyruvate synthase catalyzes the reaction: pyruvate + CoA leads to CO2 + acetyl-CoA + 2E. Electron acceptors which function with this enzyme are FAD, FMN, riboflavin, ferredoxin, and methyl viologen, but not NAD or NADP. The amebal acetate thiokinase catalyzes the reaction acetyl-CoA + ADP + Pi leads to acetate + ATP + CoA. For this apparently new enzyme we suggest the trivial name acetyl-CoA-synthetase (ADP-forming). Extracts from axenic amebae do not contain hydrogenase, but extracts from cells grown with bacteria do. It is postulated that in bacteria-grown amebae electrons generated at the pyruvate synthase step are utilized anaerobically to produce H2 via the hydrogenase and that the acetyl-CoA is converted to acetate in an energy-conserving step catalyzed by amebal acetyl-CoA synthetase. Aerobically, cells grown under either regimen may utilize the energy-conserving pyruvate-to-acetate pathway since O2 then serves as the ultimate electron acceptor.