Hyperspectral imaging for identification of irregular-shaped microplastics in water.

In this article, we demonstrate detection and identification of ten microplastic types directly in a water sample using an identification table derived from microplastic hyperspectral images. We selected a total of fourteen wavelengths which can be used to distinguish these ten microplastic types. We enhanced the visibility of these wavelengths by computationally removing water and baseline correcting with reflectance at 1550 nm. This method avoids, prevents, and eases most of the laborious sample preparation mandatory prior to analysis with robust techniques such as Raman spectroscopy and Fourier transform infrared spectroscopy. The ten different plastics were studied in water, first separately and then in a mixture. The microplastic concentrations varied depending on microplastic type and were kept <12 mg/ml per type. Finally, detection and identification were confirmed pixel-wise in a hyperspectral image of a realistic water matrix simulant including mixtures of only a few microplastic particles. All measurements have been performed with microplastics of different sizes and irregular shapes made in-house by milling commercial pellets and sheets. It enabled the establishment of a procedure for the identification of these vicious particles in real water samples. The present measurement setup of hyperspectral imaging and method of data analysis of a mixture of microplastics directly from a water-based sample may open a path towards fast, reliable, and on-site detection.

Influence of the marine feeding area on the muscle and egg fatty-acid composition of Atlantic salmon Salmo salar spawners estimated from the scale stable isotopes.

Fatty acids in muscle tissue and eggs of female Atlantic salmon Salmo salar spawners were analysed to evaluate the dietary quality of their final feeding areas in the Baltic Sea. The final likely feeding area was identified by comparing stable carbon and nitrogen isotope composition of the outermost growth region (final annulus) of scales of returned S. salar with that of reference S. salar caught from different feeding areas. Some overlap of stable-isotope reference values among the three areas, in addition to prespawning fasting, decreased the ability of muscle tri-acylglycerols to discriminate the final likely feeding area and the area's dietary quality. Among three long-chained polyunsaturated fatty acids, docosahexaenoic acid (DHA; 22:6n-3), eicosapentaenoic acid (EPA; 20:5n-3) and arachidonic acid (ARA; 20:4n-6), the proportions of ARA in total lipids of spawning S. salar muscle and eggs showed a significant negative correlation with increasing probability of S. salar having returned from the Baltic Sea main basin (i.e. the Baltic Sea proper). The results suggest that ARA in muscle and eggs is the best dietary indicator for dietary characteristics of final marine feeding area dietary characteristics among S. salar in the Baltic Sea.

The influence of timing on the effectiveness of epidural blood patches in parturients.

BACKGROUND: Postdural puncture headache is common in parturients following lumbar puncture. If headache is severe and persistent, an epidural blood patch is recommended. In this paper we reviewed the efficacy of epidural blood patches over a 13-year period at two hospitals in Finland with a particular emphasis on its timing. METHODS: The hospitals' databases were searched to identify parturients who underwent an epidural blood patch from March 1998 to June 2011. Parturients' records were reviewed to establish the characteristics and associated symptoms of headache and the effectiveness of the epidural blood patch. RESULTS: A total of 129 parturients received 151 epidural blood patches. These followed spinal (n = 49), epidural (n = 47) or combined spinal-epidural blocks (n = 33). The success rate of the first procedure was 89%, with permanent relief in 76%. The first procedure provided permanent relief of postdural puncture headache for 86% of 78 patients having the procedure after 48 h, compared to 65% of 37 patients when it was performed between 24 and 48 h, and 50% of 14 patients with the procedure within the first 24 h after dural puncture (P = 0.003). A second procedure was performed for 22 parturients due to incomplete relief (n = 5) or recurrent symptoms (n = 17); all had complete resolution of symptoms. CONCLUSIONS: Epidural blood patch performed later than 48 h following lumbar puncture or accidental dural puncture is effective in parturients with postdural puncture symptoms. The recurrence rate of symptoms after an initially successful epidural blood patch is high, and therefore patients should be provided with counselling and contact information.

Biochemical and growth acclimation of birch to night temperatures: genotypic similarities and differences.

The responses of plants to environmental factors are connected to the time of day. In this study, silver birch (Betula pendula) was grown in growth chambers at five different night temperatures (6-22 °C), using gradual changes during the evening and morning hours. Despite the increased night respiration and unaffected daytime net photosynthesis (per square metre), the carbon uptake (biomass) of birch did not decrease, probably due to enhanced biochemical processes on warmer nights and the advantage of higher temperatures during the evening and morning hours. The plant stem height, internode length, stem dry weight (DW), stem mass fraction and specific leaf area increased with warmer night temperatures. Changes in growth and metabolite concentrations were partly nonlinear along the temperature gradient. Thus, the temperature effect depends on the temperature window considered. Genotypes had both common and genotype-specific biochemical responses to night temperatures. The common responses among genotypes were related to growth responses, whereas the unique responses may indicate genotype-specific differences in acclimation. The differences in genotypic growth and metabolite levels are valuable for assessing genotype qualities and understanding the connections between the metabolome and growth.

Elevated water temperature impairs fertilization and embryonic development of whitefish Coregonus lavaretus.

The adverse effects of high temperatures on the early life stages of anadromous whitefish Coregonus lavaretus were experimentally examined by assessing fertilization success, the percentage of developmental abnormalities, cumulative mortality and the rate of embryogenesis across a range of temperatures. Temperatures >or= 7 degrees C increased the proportion of unfertilized and abnormally dividing eggs, deformed embryos and consequent mortality. The higher the temperature, the more severe were the effects. When eggs were fertilized and constantly incubated at various temperatures, the effective level for 50% of the eggs and embryos (EL50) of temperature was 7.6 degrees C at the developmental stage when eye pigmentation was visible. Fewer developmental abnormalities and a lower cumulative mortality rate were observed when embryos were exposed to high temperatures from the later, gastrula stage, than from fertilization or the four-cell stage. Irrespective of retarded development in terms of day-degrees (i.e. the sum of daily mean temperatures), a high incubation temperature reduced the development time of C. lavaretus, leading to earlier hatching, and hatched fry were shorter than at the reference temperature of 4-5 degrees C. Global warming will particularly pose risks for stenothermic species such as C. lavaretus, with early life stages being especially susceptible. Thus, relatively small increases and fluctuations in river water temperatures during the spawning season of this anadromous species may have substantial negative impacts on its recruitment and population persistence.

Starch in plasterboard sustains Streptomyces californicus growth and bioactivity of spores.

AIMS: The effects of plasterboard composition on Streptomyces californicus growth and bioactivity of spores were studied. METHODS AND RESULTS: Streptomyces californicus was grown on 13 modified plasterboards under saturated humidity conditions. The total content of fatty acid methyl esters was used for quantifying S. californicus biomass, while the spore-induced cytotoxicity and production of nitric oxide (NO), tumour necrosis factor-alpha, and interleukine-6 (IL-6) in mouse macrophages was used to assess the bioactivity of spores. Removal of starch completely from the plasterboard or only from the core reduced significantly the biomass production and the biological activity of spores in comparison with reference board. The biocide added into the core or on the liner decreased the growth markedly and inhibited the sporulation totally. The biomass production correlated positively with the spore number, cytotoxicity, and production of NO and IL-6. CONCLUSIONS: Streptomyces californicus grew under nutrient limitation on all studied plasterboards. The starch is the major factor enabling S. californicus to grow and to produce biologically active metabolites on plasterboard. SIGNIFICANCE AND IMPACT OF THE STUDY: The composition of building material has an impact on microbial growth and bioactivity of spores which may be involved in complex mechanisms leading to respiratory symptoms in the occupants in moisture damaged buildings.

Microbial community structure in biofilms and water of a drinking water distribution system determined by lipid biomarkers.

The development of microbial communities in biofilms of a drinking water distribution system was monitored, and compared to the microbial communities in water. The microbial communities were studied by phospholipid fatty acid (PLFA) profiles. In drinking water samples the most common PLFAs, with the proportion of 60.9%, were monoenoic fatty acids, such as 16:1omega7c and 18:1omega7c, indicating high abundance of gram-negative bacteria. Instead, in biofilm samples saturated fatty acids, such as 16:0 and 18:0, indicating general biomass, accounted for 54.9-78.4% of the total PLFAs. The proportions of monoenoic fatty acids in biofilm increased from 11.5% to 31.2% with water aging from 22 h to 62 h in the distribution system. In conclusion, water aging affected the structure of microbial communities in biofilms, and the microbes in water differed from those in biofilms. These differences might also reflect the differences in the physiological state of the microbes, which is influenced by water chemistry and by the growth environment, i.e. water or biofilm.

Gas chromatographic-mass spectrometric detection of 2- and 3-hydroxy fatty acids as methyl esters from soil, sediment and biofilm.

Hydroxy fatty acids (OH-FAs) can be used in the characterization of microbial communities, especially Gram-negative bacteria. We prepared methyl esters of 2- and 3-OH-FAs from the lipid extraction residue of soil, sediment, and biofilm samples without further purification or derivatization of hydroxyl groups. OH-FA methyl esters were analyzed using a gas chromatograph equipped with a mass selective detector (GC-MS). The ions followed in MS were m/z 103 for 3-OH-FAs and m/z 90 and M-59 for 2-OH-FAs. The rapid determination of 3- and 2-OH-FAs concomitantly with phospholipid fatty acids provided more detailed information on the microbial communities present in soil, sediment, and drinking water biofilm.

Herbivore-induced allene oxide synthase transcripts and jasmonic acid in Nicotiana attenuata.

Exogenous jasmonate treatment of Nicotiana attenuata Torr. ex Wats. plants elicits durable resistance against herbivores and attack from its specialist herbivore, Manduca sexta, results in an amplification of the transient wound-induced increase in endogenous jasmonic acid levels (JA). To understand whether this "JA burst" is under transcriptional control, we cloned allene oxide synthase (AOS; EC 4.2.1.92), the enzyme that catalyzes the dehydration of 13(S)-hydroperoxy octadecatrienoic acid to an allene oxide, the first specific reaction in JA biosynthesis. An AOS cDNA coding for a 520 aa protein (58.6 kDa) with an isoelectric point of 8.74 was overexpressed in bacteria and determined to be a functional AOS. Southern blot analysis indicated the presence of more than one gene and AOS transcripts were detected in all organs, with the highest levels in stems, stem leaves and flowers. Attack by M. sexta larvae resulted in a sustained JA burst producing an endogenous JA amount 9-fold above control levels and 3-fold above maximum wound-induced levels, a response which could be mimicked by the addition of Manduca oral secretion and regurgitant to puncture wounds. M. sexta attack, wounding and regurgitant treatment transiently increased AOS transcript in the wounded leaf, but increases were not proportional to the JA response. Moreover, transcript accumulation lagged behind JA accumulation. Systemic wound-induced increases in AOS transcript, AOS activity or JA accumulation could not be detected. We conclude that increase in AOS transcript does not contribute to the initial increase in endogenous JA, but may contribute to sustaining the JA burst.

Rapid HPLC screening of jasmonate-induced increases in tobacco alkaloids, phenolics, and diterpene glycosides in Nicotiana attenuata.

A rapid, HPLC-based screening procedure for the main classes of secondary metabolites in Nicotiana attenuata leaves (alkaloids, phenolics, and diterpene glycosides) is reported. In a single step, leaves are extracted in aqueous acidified (0.5% acetic acid) methanol, and the extracted compounds are separated by reversed-phase HPLC with an acidic water/acetonitrile gradient in <30 min. The utility of the method in quantifying changes in the secondary metabolites after methyl jasmonate treatment of the plants, a treatment known to elicit resistance to herbivores in nature, is illustrated. Methyl jasmonate treatment elicited dramatic increases in some secondary metabolites (caffeoylputrescine, nicotine, and diterpene glycosides increased 12.5-, 1.4-, and 1.9-fold, respectively) but left others, such as rutin, unchanged. Such broad-based analytical screens will help characterize environmental and genetic changes in secondary metabolite profiles.

Cooperation between mental health professionals and doctors in a Balint-oriented supervision group.

A Balint-oriented supervision group for physicians is described concentrating on the study of the patient-doctor relationship, the recognition and diagnosis of psychiatric problems, and the planning of psychiatric treatment. The group includes five general practitioners, a gynecologist, a dermatologist, a psychiatrist and a psychologist, who have met once a month for an hour over a period of 12 years. Interaction between the physicians and the mental health professionals is illustrated by two clinical examples. The group helps the physician recognize, tolerate and use his countertransference feelings, and facilitates the examination and treatment of patients suffering from psychiatric problems. In Balint-oriented group work, the focus can be moved from physical symptoms to include observation of the patient's emotional life and significant object relations, to the factors that are crucial for his psychological balance. This kind of holistic observation in the examination and treatment of psychiatric problems is as important as appropriate laboratory investigations in the diagnosis and care of physical diseases.

The evolution of the internal dialogue during the psychoanalytic psychotherapy process.

This article illustrates curative changes occurring in psychoanalytic psychotherapy by developing the internal dialogue in the mind. By the internal dialogue, I mean the internalization of the therapy process during the recurrent therapy sessions so that the external dialogue between therapist and patient becomes the corresponding internal structure of the patient. In this way, there is a development of the patient's capacity to identify himself and the therapist as separate, sentient, thinking, and reflecting individuals, who have a free internal world of their own. The evolution of the internal dialogue takes place by gradually progressing symbolization achieved through a four-step symbolization-reflectiveness approach. This process is one of the specific curative factors in the psychotherapeutic treatment of borderline patients. The psychodynamics are illustrated by material from one session each of the early and the final stages of therapy, showing a shift from monologue to internal dialogue. The evolved internal dialogue is a central part of the patient's budding thinking capacity, which creates a ground for her subjectivity and autonomy. This study underlines and specifies three factors as cornerstones of the evolution of identity in psychotherapy: symbolization, reflectiveness, and the internal dialogue.

Ion regulation in whitefish (Coregonus lavaretus L.) yolk-sac fry exposed to low pH and aluminum at low and moderate ionic strength.

Anadromous whitefish [Coregonus lavaretus (L.)] were exposed during the yolk-sac phase to combinations of pH values of 6.0, 5.5, 5.0, 4.5, and 4.0 and nominal Al concentrations of 0, 100, 200, and 400 micrograms liter-1 for 4 days. The test waters were: (1) lake water diluted 1:1 with ion exchanged water (DLW) and (2) artificial water (AW). The major differences were: [Ca2+] 0.069 mmol liter-1 in DLW versus 0.034 mmol liter-1 in AW, [Na+] 1.5 and [Cl-] 1.8 times higher in DLW, with no dissolved organic material in AW. The fry were sampled daily for the analysis of exchangeable body Na+ and Cl- contents, and the swimming activity and mortality were registered. Aluminum initially protected yolk-sac fry against acidic water at pH 4.0 both in DLW and in AW. Generally, however, an increase in [Al] and a decrease in pH increased the adverse effects, seen as decreased exchangeable body Na+ and Cl- content, decreased swimming activity, and increased mortality. The effects of Al were more pronounced in AW compared with DLW. In AW, the Na+ and Cl- content of the fry already decreased after 1 day of exposure to a level that predicts mortalities. There was a time trend in DLW toward decreasing exchangeable body Na+ and Cl- concentrations as a function of increasing acidity and [Al]. In DLW, mortalities occurred primarily at pH 4.0; at that pH value in AW, the mortality rate was even higher, and mortalities also occurred after 2 to 4 days at all other pH values. There was a threshold limit in exchangeable body Na+ and Cl- concentrations at which yolk-sac fry became passive; it was at approximately 0.3 mumol per fry in DLW and was slightly higher in AW. In the DLW control with moderate ionic strength, the Na+/Cl- ratio was above 1 from the second exposure day on but remained closer to 1 in soft AW control. The Na+/Cl- ratio was predominantly > 1 in AW when the fry were exposed to moderate acidities but approached 1 at pH values < 5.0. The Na+/Cl- ratio was mostly < or = 1 in DLW, irrespective of the exposure, indicating cation selectivity of ion leakage pathways in slightly harder water.

Genetic aberrations in pediatric acute lymphoblastic leukemia by comparative genomic hybridization.

Classical cytogenetic analysis plays an important role in the diagnosis and classification of childhood acute lymphoblastic leukemia (ALL). However, poor in vitro growth of the malignant cells and suboptimal quality of metaphase spreads may sometimes cause false-negative findings (normal karyotype). We used comparative genomic hybridization (CGH) to study whether this new method is able to detect and characterize genetic aberrations not detected by karyotyping. CGH showed clonal genetic aberrations in 8 of 13 cases, most of which showed gains of several chromosomes, indicating hyperdiploidy. The sensitivity of CGH was sufficient to detect a small interstitial deletion of 6q. One karyotypically complex case was resolved by CGH showing a high-level amplification of DNA sequences originating from the 12p12-13. Interphase fluorescence in situ hybridization (FISH) analyses confirmed the CGH findings in 2 cases, validating the accuracy of CGH. In conclusion, CGH experiments established the known fact that hyperdiploidy is the most common finding in pediatric ALLs and that CGH may detect aberrations that are not seen in the G-banded karyotype. CGH was also able to further characterize genetic aberrations such as gene amplification, which is occasionally involved in pediatric ALL as well as in other leukemias.

A transcript map of the newly defined 165 kb Wolf-Hirschhorn syndrome critical region.

Wolf-Hirschhorn syndrome (WHS) is a multiple malformation syndrome characterised by mental and developmental defects resulting from the absence of a segment of one chromosome 4 short arm (4p16.3). Due to the complex and variable expression of this disorder, it is thought that the WHS is a contiguous gene syndrome with an undefined number of genes contributing to the phenotype. In an effort to identify genes that contribute to human development and whose absence results in this syndrome, we have utilised a series of landmark cosmids to characterise a collection of WHS patient derived cell lines. Fluorescence in situ hybridisation with these cosmids was used to refine the WHS critical region (WHSCR) to 260 kb. The genomic sequence of this region is available and analysis of this sequence through BLAST detected several cDNA clones in the dbEST data base. A total of nine independent cDNAs, and their predicted translation products, from this analysis show no significant similarity to members of DNA or protein databases. Furthermore, these genes have been localised within the WHS critical region and reveal an interesting pattern of transcriptional organisation. A previously published report of a patient with proximal 4p- syndrome further refines the WHSCR to 165 kb defined by the loci D4S166 and D4S3327. This work provides the starting point to understand how multiple genes or other mechanisms can contribute to the complex phenotype associated with the Wolf-Hirschhorn syndrome.

The meaning of the symbolic function in psychoanalytic psychotherapy: clinical theory and psychotherapeutic applications.

This study presents the clinical theory and psychotherapeutic applications of symbolic function in psychoanalytic psychotherapy. The three modes of symbolic function (indexical, iconic and conventional symbolic) form the representations of the self and the object and further the unconscious fantasy, which includes the affective bond between these representations. During his/her development the child absorbs from the mother a new ego capacity (by means of identification), which I call the reflective-integration capacity. Then, the ego of the child has two crucial functions: the fantasy world adopted through the symbolic function and the reflective-integrative capacity. In psychoanalytic psychotherapy our aim is to study the fantasy world of the patient by means of the reflective-integrative capacity, or if this capacity is missing or weak, to promote its development. During psychoanalytic psychotherapy a shared area of reflection-integration is formed, in which the healing changes occur. These healing changes include mainly the formation, consolidation and enrichment of the symbolic function of the patient. The consolidation of symbolic function and the capacity to move within the different modes of symbolic function occur in the area of psychology in which there are incapabilities and/or conflicts (e.g. separation anxiety) in the patient. Clinical case material is presented to illustrate these phenomena.

Maternal origin of transferrin receptor positive cells in venous blood of pregnant women.

We studied the origin of transferrin receptor (CD71) positive cells in blood from seven women pregnant with a male fetus in order to explore if fetal cells could be detected among them. We used a technique that allows direct chromosomal analysis by in situ hybridization on immunologically and morphologically classified cells. Enrichment was performed by magnetic activated cell sorting (miniMACS) using an anti-CD71 monoclonal antibody. The cells were immunophenotyped by alkaline phosphatase anti-alkaline phosphatase immunostaining with the same antibody. The origin of the immunophenotyped cells was studied by in situ hybridization using an X cosmid Y repeat chromosome specific probe cocktail. CD71 positive cells were found in six of the seven women at the range of 4 to 43 in respective samples. Over 90% of the CD71 positive cells were nucleated erythrocytes. None of the detected positive cells were shown to be fetal. Thus, the use of transferrin receptor antigen alone in combination with the miniMACS may not be sufficient for enrichment of fetal cells.

Maternal origin of nucleated erythrocytes in peripheral venous blood of pregnant women.

We studied the origin of nucleated red blood cells (NRBC) in peripheral venous blood samples from 40 pregnant women carrying a male fetus, using a technique that allows direct chromosomal analysis by in situ hybridisation on immunologically and morphologically classified cells. Samples from ten nulligravid women were studied as controls. NRBC were enriched by negative magnetic activated cell sorting (miniMACS) using anti-CD45 monoclonal antibody. NRBC were detected by alkaline phosphatase anti-alkaline phosphatase immunostaining using a monoclonal anti-glycophorin A antibody. The origin of the NRBC was determined by fluorescence in situ hybridisation using X and Y specific probes. NRBC were found in 37 of the 40 pregnant women at a range of 1 to 230 per 20 ml of venous blood and in 6 of the 10 controls at a range of 1 to 3 per 20 ml of venous blood. All NRBC detected in the pregnant women were evidently of maternal origin, and in the pregnant women the number of NRBC was significantly higher (P < 0.05) than in the controls. Pregnancy per se seems to induce the appearance of maternal NRBC in the circulation, and it cannot therefore be assumed that NRBC isolated from the maternal blood are of fetal origin on the basis of morphology alone. Discrimination of fetal NRBC must occur for prenatal diagnosis of fetal genetic disorders.

ISH and PCR study with Y-specific probe/primers.

Our main aim was to evaluate whether maternal whole venous blood could be used for determination of fetal sex, when no enrichment of fetal cells was attempted and when "standard" interphase cytogenetics and PCR analysis were adopted. Altogether 39 pregnant women were studied by using ISH and 59 by using PCR. Out of the 59 pregnant women, 26 carried a male fetus and 33 a female fetus. By ISH, Y-positive cells were detected in 12 of 19 pregnancies with a male fetus and in two of the 20 pregnancies with a female fetus. The frequency of the fetal cells ranged from 1 in 639 to 1 in 100,000. By nested PCR with primers flanking a Y-specific repeat sequence, the positive band indicating a male fetus was found in one of the 26 pregnancies with a male fetus and in one of the 33 pregnancies with a female fetus. According to our results, fetal cells in maternal blood cannot be reliably used for prenatal diagnosis without enrichment of fetal cells.