The effects of prenatal, perinatal and neonatal factors on academic performance in primary school age children.

OBJECTIVE: To synthesize existing evidence about the effects of prenatal, perinatal and neonatal factors on academic performance in primary school age children (up to 12 years). DESIGN: Review article. SETTING: National Institute of Mental Health, Klecany. METHODS: Literature review using databases SCOPUS, WoS, ProQuest, MEDLINE, PsycArticles, PsycINFO and ERIC with a search strategy (academ* OR schol* OR school*) AND (success* OR aptitude OR fail* OR abilit* OR perform* OR achiev* OR progress* OR outcom* OR attain*) AND (pregnan* OR prenatal* OR perinatal* OR childbirth OR (child AND birth) OR birth OR intrapart* OR postpart* OR preterm OR pre-term OR premature OR (low AND birth AND weight) OR cesarean OR caesarean OR c-section OR (c AND section) OR breastfeeding OR (breast AND feeding)) AND (child* OR kid* OR pupil*). Only peer-reviewed studies in English were included, no time limits for publication date were set. RESULTS: The main factors influencing academic performance include prematurity and factors related to maternal health and life style in pregnancy (substance abuse, health complications such as diabetes mellitus and hypothyroidism, psychiatric diagnosis, use of antiepileptic drugs, dietary habits, especially restriction of caloric intake, exposure to harmful influences as heavy metals, ionizing radiation and stress). Academic performance has also been found to be associated with delivery mode and several neonatal factors, particularly respiratory or cardiac insufficiency and infection. CONCLUSION: Factors related to pregnancy, childbirth and the early postpartum period may have a negative impact on school success in children. Their early detection can help to develop strategies for early support and intervention in vulnerable groups of children.

Psychosocial risk factors for emergency cesarean section.

OBJECTIVE: To compare the effects of childbirth fear and trait anxiety on the risk of emergency cesarean section; to analyze whether emergency cesarean section is associated with low mastery and maternal self-esteem; to examine whether stress and low social support in pregnancy may be considered risk factors for emergency cesarean section. DESIGN: Original study. SETTING: Department of Psychology, Faculty of Arts, Charles University, Prague. METHODS: The study sample consisted of 447 women who gave birth at a maternity hospital in Vysočina Region (Havlíčkův Brod, Jihlava, Pelhřimov, Třebíč, Nové Město na Moravě) between October 2013 and September 2014. In the last trimester of pregnancy, the women completed validated questionnaires designed to assess fear of childbirth, general anxiety, maternal self-esteem, mastery, perceived stress and social support. Data regarding the course of labor were extracted from medical records. The association between the psychosocial factors and the risk of delivery via emergency cesarean section was analyzed using multiple logistic regression adjusted for marital status, parity, childs sex and epidural anesthesia. RESULTS: A total of 73 women (16.3%) delivered by emergency cesarean section. The only statistically significant psychosocial predictor of emergency cesarean section was fear of childbirth (the women with strong fear had a twice higher risk; OR = 2.01; p = 0.021), whereas low maternal self-esteem was marginally significant (OR = 1.68; p = 0.082) in the adjusted analysis. No association between emergency cesarean section and general anxiety, mastery, stress or social support in pregnancy was found. The risk of cesarean section was higher for primiparous women and lower for women who gave birth to a girl. CONCLUSION: Fear of childbirth but not general anxiety is associated with a higher risk of emergency cesarean section. The women who experience strong fear of childbirth during pregnancy should be recommended to attend antenatal classes or, in case of extremely severe childbirth fear, to seek psychological counseling.

[Are womens attitudes towards pregnancy, childbirth and motherhood associated with length of labour?] / Souvisí postoje zen k tehotenství, porodu a materství s délkou porodu?

OBJECTIVE: To examine the association between length of labour and womens attitudes towards pregnancy, childbirth and motherhood, while controlling for biomedical and maternal psychosocial characteristics. DESIGN: Original study. SETTING: Department of Psychology, Faculty of Arts and Philosophy, Charles University, Prague. METHODS: The sample consisted of 362 women who gave birth in one of the five maternity hospitals in Vysocina region (Havlíckuv Brod, Jihlava, Pelhrimov, Trebíc, Nové Mesto na Morave) between October 2013 and September 2014. The data were collected at two points in time. In the third trimester of pregnancy, womens attitudes towards pregnancy, childbirth and motherhood and other psychosocial characteristics were surveyed using the range of validated tools. Within one week of delivery, women were asked about social support they received during labour. Data concerning the course of labour were extracted from medical records. The dependent variable was the active phase of the first stage of labour (time from 3 cm to full dilatation). The differences among women with prolonged/rapid labour (duration above 90th percentile/less than 10th percentile) with respect to their attitudes towards pregnancy, childbirth and motherhood and other characteristics were assessed using the Students t test and the χ2 test. Factors associated with length of labour were analysed using logistic regression. The results were adjusted for maternal age, marital status and newborn weight. RESULTS: In labours without oxytocin administration, the median of the duration of active phase of the first stage was 180 minutes for primiparas and 144 minutes for multiparas. Women with prolonged labour reported more frequently low maternal self-esteem compared to women with normal labour duration (34.0% vs. 19.1%; p = 0,021), but maternal self-esteem was not a significant predictor of prolonged labour in regression analysis. No other differences were found with respect to psychosocial and personality characteristics in women with prolonged and normal labour. Women with rapid labour did not differ from women with normal labour as for their attitudes towards pregnancy, labour and motherhood. Nevertheless, they reported clinically relevant scores for trait anxiety more frequently, a difference that approached statistical significance (52.6% vs. 38.3%; p = 0,090). Multiple regression analysis showed a trend towards trait anxiety being associated with rapid labour (p = 0,098). No significant predictor of labour duration was identified except newborn weight which predicted both prolonged and rapid labour. CONCLUSION: Our data indicate that the association between length of labour and womens attitudes towards pregnancy, childbirth and motherhood is rather weak and clinically less relevant. Our results portray the newborn weight as a key factor affecting labour duration.

[Postpartum blues - a Czech adaptation of the Maternity Blues Questionnaire]. / Poporodní blues - ceská adaptace dotazníku "Maternity Blues Questionnaire".

OBJECTIVE: To validate the Kennerley and Gaths Maternity Blues Questionnaire (MBQ) for the Czech postpartum population, to present the psychometric properties of the Czech version of that screening method, and to assess its predictive power for the risk of postpartum depression. DESIGN: Original study. SETTING: Department of Psychology, Faculty of Arts and Philosophy, Charles University, Prague. METHODS: The Czech version of the MBQ was validated on a sample of 1093 women. The data were collected from October 2013 to September 2014 at all maternity hospitals in Vysocina region. The MBQ was administered on a one-time basis during womens postpartal stay at maternity hospital. After six weeks post partum, a screening for postpartum depression was performed using the Edinburgh Postnatal Depression Scale (EPDS). The cut-off point was set at 10/11 for MBQ and 12/13 for EPDS as such were the respective levels achieved by the 90th percentile in the MBQ and EPDS scores. The sociodemografic data were collected at the time of completing the MBQ. A logistic regression was performed to identify the predictors of severe blues. Cronbachs alpha was calculated to assess the internal consistency of the MBQ as a whole and its component scales. In order to assess the validity of the MBQ, a logistic regression was used to analyze the association between the MBQ and EPDS scores. The norms for the Czech version of MBQ are presented as percentiles. RESULTS: The MBQ scores showed a gradual rise over the days following the delivery (day 0 to day 4). The percentage of women with severe blues (MBQ score > 10) increased from 7.3% to 14.55% between day 0 and day 4. The most frequent feelings and mood states experienced by women in the first postpartum days included tiredness (61%), decreased self-confidence (30%), over-sensitivity (26%) and tension (19%), while 6,5% of women felt low spirited and 7% felt depressed. The women suffering from severe blues reported most frequently the same states of mood as did the women in the whole sample, but the rates of those states were higher: 83% for tiredness, 81% for decreased self-confidence, 79% for over-sensitivity, 71% for tension, while 46% of women with severe blues felt low spirited and 51% felt depressed. The significant risk factors for severe blues included parity (multipara, OR = 0.42; p < 0,001), mode of delivery (reference category unassisted vaginal delivery: emergency sectio caesarea, OR = 2.188, p = 0.004; planned sectio caesarea, OR = 1.843, p = 0.03; assisted vaginal delivery, OR = 6.136; p < 0.001), and previous depression (OR = 4.71, p = 0.003). Cronbachs alpha of the individual scales ranged from 0.34 to 0.76, Cronbachs alpha for the MBQ as a whole was 0.88. The severe blues were found to be a predictive factor for postpartum depression as assessed with EPDS (OR = 5.90; p < 0.001). CONCLUSION: With its high reliability and validity, the MBQ appears to be a useful tool for clinical practice and research. The MBQ can be used to identify the women with severe blues and with an increased risk of postpartum depression.

[Satisfaction with perinatal care in Vysocina region in the period between October 2013 and September 2014]. / Spokojenost zen s perinatální pécí v Kraji Vysocina za období ríjen 2013 az zárí 2014.

OBJECTIVE: To assess women's satisfaction with perinatal care provided in maternity hospitals in Vysocina region, to identify the areas with high satisfaction scores as well as those requiring improvement, and to describe the factors influencing women's satisfaction, i. e. dissatisfaction with the care provided during labor and birth and the early postpartal period. DESIGN: Original study. SETTING: Department of Psychology, Faculty of Philo-sophy, Charles University, Prague. METHODS: The satisfaction survey was conducted in all maternity hospitals in Vysocina region (Jihlava, Havlíckuv Brod, Trebíc, Pelhrimov, Nové mesto na Morave) during the period between October 2013 and September 2014. All women who had given birth in those hospitals during the period were approached and asked to participate in this survey. The women evaluated the perinatal care not before 58 days after birth, so that the evaluation of perinatal care did not take place directly during their stay at maternity hospital. In total, 1366 women took part in the study. The original Czech questionnaire KLI-P was used for the data collection. The KLI-P measures psychosocial climate of maternity hospitals on the following six scales: helpfulness and empathy of caregivers; control and involvement in decision-making; communication of information and availability of caregivers; dismissive attitude and lack of interest; physical comfort and services. The satisfaction rates with component dimensions of intrapartal and postpartal care at the maternity hospitals in Vysocina region were compared to the satisfaction rates for the Czech Republic as a whole as obtained in our previous study. We used the Kruskal-Wallis test for this comparison. The ordinal logistical regression (cumulative logit model) was used to identify predictors of women's satisfaction with intrapartum and postpartum care in Vysocina region. RESULTS: The women who delivered at maternity hospitals in Vysocina region were significantly more satisfied with all dimensions of care received both at delivery (DU) and after-birth unit (ABU) as compared to the average parturient's satisfaction in the Czech Republic as specified in our previous study. The best rated scale at DU in Vysocina region was Physical comfort and services (93% vs. 85% for the whole country; P < 0,0001), while the worst evaluation score received the scale Control and involvement in decision-making (75% vs. 58% for the whole country; P < 0,0001). At ABU, the best rated scale was Control and involvement in decision-making (95% vs. 89% for the whole country; P < 0,0001), while the lowest evaluation score was found for the scale Physical comfort and services (85% vs. 76% for the whole Republic; P < 0,0001). The women perceived generally the caregivers as helpful and kind but they often missed sufficient emotional support. They considered the information received easy to understand, but they would welcome more information in general. Among the most important determinants of parturients satisfaction with care at DU were kind and helpful attitude of midwives (P < 0,0001), confidence in physicians (P < 0,0001), sufficient provision of information (P = 0,0016), pleasant appearance of the delivery room (P = 0,0010), kind and helpful attitude of physicians (P = 0,018). Among the most important determinants of satisfaction with care at ABU were sufficient provision of information (P = 0,0007), equipment of the room (P = 0,0014), informations regarding the care for the newborn (P = 0,0013), emotional support (P = 0,0039) and confidence in staff employed in newborn care (P = 0,025). CONCLUSION: Satisfaction with care received at maternity hospitals in Vysocina region was rated very high. However, despite its high quality, the perinatal care in Vysocina region suffers from several shortcomings. Regarding the significance of the individual determinants of parturient's satisfaction, improving the quality of care requires first of all better provision of information both during the labor and postpartal period. It would be beneficial to provide the health care professionals with the opportunities of attending a training in breastfeeding support and communication and of taking part in clinical supervision.

Self-sustaining reactions as a tool to study mechanochemical activation.

Ever since Faraday reported on the reduction of AgCl with Zn and Sn the "dry way", the study of Mechanically induced Self-propagating Reactions (MSRs) has been an important area within mechanochemistry. An interesting phenomenon is the mutual suppression of ignition in some mixed metal-chalcogen systems, such as in (1 - x)(Sn + Se) + x(Zn + Se) powders. Here both the Sn + Se and Zn + Se mixtures react in a self-sustaining manner after some activation time, but when they are combined, the reaction is gradual in the middle of the concentration scale. Mechanically induced metal-chalcogen combination reactions were studied by Chakurov et al. in the 1980s, using a low-energy vibratory mill. Similar measurements were carried out in our laboratory using the more energetic SPEX 8000 shaker mill. The results show qualitative similarities, but the details are different. It is suggested that the loss of MSR in mixed systems is the consequence of the very different properties of the binary systems, so that either one of the components (Zn) or a product formed gradually without ignition (e.g. SnSe) can act as an inert component relative to the rest of the system. Several examples are presented and the effect of the milling conditions is discussed. Finding new systems with similar behaviour and detailed studies of the activated state are needed to understand MSR in these systems.

[Psychosocial climate in maternity hospitals from the perspective of parturients II. Predictors of womens satisfaction with perinatal care in the Czech Republic]. / Psychosociální klima porodnice ocima rodicek II. Prediktory spokojenosti s perinatální pécí v Ceské republice.

OBJECTIVE: To identify the key determinants of parturients satisfaction with care provided in the Czech maternity hospitals and to draw up recommendation for good practice. DESIGN: Original study. SETTING: Department of Psychology, Faculty of Philosophy, Charles University, Prague. METHODS: Statistical analysis of data from the representative sample of Czech parturients (n = 1195) who gave birth from 2005 to 2012 in Czech maternity hospitals was conducted. Ordinal logistical regression (cumulative logit model) was used to identify predictors of womens satisfaction with intrapartum and postpartum care and their willingness to return and give birth in the same hospital as well as to recommend the hospital to others. The independent variables were the individual items of the KLI-P questionnaire measuring psychosocial climate in maternity hospitals and socio-demographic and anamnestic characteristics of parturients. RESULTS: Among the most important determinants of parturients satisfaction with care at delivery unit were well-timed provision of information concerning a treatment plan (OR = 2,79; p < 0,0001), privacy during the first stage of labour (OR = 2,81; p < 0,0001), kind and helpful attitude of physicians (OR = 2,67; p < 0,0001) and confidence in physicians (OR = 2,68; p = 0,0001). Among the most important determinants of satisfaction with care at after-birth unit were well-timed provision of information concerning a treatment plan (OR = 2,25; p < 0,0001), active breastfeeding support (OR = 2,19;p < 0,0001), availability of caregivers (OR = 2,16;p < 0,0001), multiparity (OR = 1,52; p = 0,0001), respect shownby staff employed in mother care (OR = 2,02; p = 0,0004) and kind and helpful attitude of staff employed in newborn care (OR = 1,86; p = 0,0005). When considering the future place of birth and recommendation of the hospital to others, women predominantly take into account the care provided at delivery unit. Intrapartal care evaluation was affected to a lesser degree by satisfaction with midwives as compared to satisfaction with physicians. CONCLUSION: Analyses of the most important predictors of satisfaction with perinatal care indicate the need for practice to focus on enhancing psychosocial competences (particularly communication skills) of health care providers, more active breastfeeding support and communication of consistent information (especially concerning newborn care) by caregivers at after-birth unit.

[Psychosocial climate in maternity hospitals from the perspective of parturients I. Results from a national survey on perinatal care satisfactionusing a representative sample of 1195 Czech parturients]. / Psychosociální klima porodnice ocima rodicek I. Výsledky celorepublikového pruzkumu spokojenosti s perinatální pécí v CR u reprezentativního souboru 1195 rodicek.

OBJECTIVE: To assess women's satisfaction with psychosocial aspects of perinatal care provided in Czech maternity hospitals, to identify areas that need improvement and to compare satisfaction with maternity care between selected subgroups of parturients. DESIGN: Original study. SETTING: Department of Psychology, Faculty of Philosophy, Charles University, Prague. METHODS: A survey on satisfaction with perinatal care was conducted with a sample of 1195 Czech parturients. The sample was representative of the Czech parturients population in terms of educational level, age, parity, and rate of vaginal and caesarean section delivery. The sample was proportionated as regards the number of births at small and large hospitals and at hospitals in different regions as well. All currently existing Czech maternity hospitals were included. For the data collection, the original Czech questionnaire KLI-P was used. The KLI-P measures psychosocial climate of maternity hospitals on following six scales: helpfulness and empathy of caregivers; control and involvement in decision-making; communication of information and availability of caregivers; dismissive attitude and lack of interest; physical comfort and services. In addition, differences in satisfaction rates between different subgroups of respondents were investigated: primiparas/multiparas, women with lower/higher educational status, women who gave birth at smaller/lager hospitals (< 800 / > 800 births per year), women who gave birth at university/other hospitals, women after vaginal delivery/caesarean section, women accommodated in high-standard rooms at after-birth unit, and women who filled the questionnaire within one year after/later than one year after delivery. RESULTS: The overall satisfaction with care provided at delivery unit (DU) and after-birth unit (ABU) was 70% and 61%, respectively. The best rated scale at DU was physical comfort and services (69%), the worst evaluation score received the scale control and involvement in decision-making (34%). At ABU, the best rated scale was control and involvement in decision-making (76%) while the lowest evaluation score was found for the scale dismissive attitude and lack of interest (48% - reverse-scored). The items with the best scores referred to the cleanliness at DU and mother-infant contact at ABU, the items with the lowest evaluation scores referred to emotional support provided by physicians at DU, involvement in decision-making concerning the position during the second stage of labour and quality of food. Significantly more satisfied with care provided at DU were multiparas, women who gave birth at non-university hospitals and women who gave birth vaginally. Significantly more satisfied with care at ABU were multiparas, women with lower educational status, women who gave birth at non-university and smaller hospitals (< 800 births per year) and women who evaluated a given hospital within one year after delivery. CONCLUSION: Despite its rather high quality, the Czech perinatal care suffers from several shortcomings as regards its psychosocial aspects. These shortcomings include lack of respect and empathy shown by caregivers, poor communication of information and low involvement of parturients in decision-making. Improving the quality of care at the Czech maternity hospitals requires empowerement of parturients in the system of perinatal care and development of psychological and psychosocial compteneces of health care providers.

[The psychosocial aspects of perinatal care and their relationship to selected medical interventions and health complications during parturition]. / Souvislost psychosociálních aspektu perinatální péce s nekterými zákroky a zdravotními komplikacemi v prubehu porodu.

OBJECTIVE: Find association between psychosocial factors of perinatal care and selected childbirth complications and interventions. DESIGN: Original study. SETTING: Department of Psychology, Faculty of Arts and Philosophy, Charles University, Prague. METHODS: Statistical data analysis of questionnaire survey of low-risk parturients (n=657) was carried out. Respondents were divided into experimental groups (EG) according to experienced interventions and complications (hemorrhage, failure to progress, fetal distress, forceps/VEX delivery, caesarean birth, failure to progress followed by caesarean birth, labour acceleration infusion, other infusion, epidural anesthesia, other form of pharmacological analgesia, episiotomy, amniotomy). EG were compared with a control group (CG) of women with no complications and interventions (n=107) in evaluation of psychosocial factors of perinatal care (healthcare provider attitude; control; communication; woman-friendliness of hospital rules) and physical comfort and services, presence of other persons at birth, prevailing emotional reaction and overall satisfaction with maternity care. The interpretation of results draws from qualitative analysis of open-ended question answers. RESULTS: EG and CG differed significantly in control (all EGs scored lower) and healthcare provider attitude evaluation (significantly more negative perception at EGs: hemorrhage, failure to progress, fetal distress, forceps/VEX delivery, labour acceleration infusion, episiotomy). Significant differences were noted also for woman-friendliness of hospital rules (lower scores for EGs: hemorrhage, failure to progress, failure to progress followed by caesarean birth) and for presence of other persons at birth. Possible interpretations of the above results are discussed. CONCLUSION: Psychosocial factors of perinatal care constitute important childbirth process determinants, while playing a key role for parturients ability to cope with anxiety and stress connected with childbirth complications and interventions. Beneficial psychosocial climate of a hospital contributes to higher patient satisfaction as well as to lower maternal and neonatal morbidity.

[The psychosocial needs of newborn children in the context of perinatal care]. / Psychosociální potreby novorozence v kontextu perinatální péce.

OBJECTIVE: To outline the variety of the psychosocial needs of newborn children to be taken into account in maternal-child nursing and to present recommendations for good practice. DESIGN: Review article. SETTING: Department of Psychology, Faculty of Philosophy, Charles University, Prague. SUBJECT AND METHOD: This study is focused on psychological importance of birth experience for the newborn, early mother-child contact, initiation of breastfeeding as well as on the psychosocial needs of the preterm newborn and newborn children at health risk. Both conclusions of scientific literature and the results of our research demonstrate that respecting the psychosocial needs of the newborn in mother-child nursing is beneficial not only for psychological development of children, but also for their physical health in short-term as well as long-term perspective. CONCLUSION: Despite its rather high quality, the perinatal health care in the Czech republic still suffers from several shortcomings in the psychological field. More emphasis needs to be put on supporting the early mother-infant contact, individual needs of parturients and their infants and initiation of breastfeeding, especially as regards women after cesarean section delivery and mothers of preterm infants.

[Psychosocial factors influencing the perinatal health care satisfaction of parturients]. / Psychosociální faktory ovlivnující spokojenost rodicek s perinatální pécí.

OBJECTIVE: To give an overview of the main psychosocial factors influencing the perinatal health care satisfaction of parturients and present recommendations for good practice. DESIGN: Original study. SETTING: Department of Psychology, Faculty of Philosophy, Charles University, Prague. METHODS: Qualitative methodology--content analysis of 189 childbirth narratives written by parturients and demi-structured interviews with 44 parturients. RESULTS: Seven dimensions of perinatal health care satisfaction of parturients were identified: 1. staff attitude and behavior; 2. staff communication; 3. parturient's participation in decision-making; 4. support of early mother-baby contact; 5. breastfeeding support; 6. mother-baby friendliness of maternity unit operation rules; 7. clarity of maternity unit operation rules. The most important dimensions include empathy and psychological support, respect for privacy and feelings of shame, relational symmetry/asymmetry and quality of provided information. CONCLUSION: Psychosocial aspects are a decisive criterion of perinatal health care satisfaction of parturients. Psychological competences of health care providers and staff make an inseparable part of their competence, with communication skills development and prevention and therapy of the burn-out syndrome deserving special attention.

Cell proliferation and apoptosis in stromal corneal dystrophies.

The aim of our study was to evaluate corneal cell proliferation and apoptosis in cases of granular, macular and lattice dystrophy, and to provide evidence which may help to clarify whether apoptosis is a pathogenic factor in any of these dystrophies. The study group comprised 39 eyes (from 33 patients) which had undergone penetrating keratoplasty (PK) for stromal dystrophies: these comprised 12 eyes (from 9 patients, 55.5% males) with granular dystrophy, 13 eyes (12 patients, 33.3% males) with macular dystrophy, and 14 eyes (13 patients, 61.5% males) with lattice type I dystrophy. A further 4 corneal buttons from enucleated eyes of 4 patients with choroideal melanoma served as controls. Immunocytochemical analysis of Ki67 (DNAcon Kit, DakoCytomation A/S, Glostrup, Denmark) was used for evaluation of cell proliferation. Apoptosis was detected by use of the TUNEL (terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick-end labelling) assay method (Apoptag reagent, Q-Biogene, Strasbourg, France). Statistical comparisons were made using the Mann-Whitney test. No Ki67-positive cells were detected in the study-group or control corneas. In control corneas no apoptotic activity was found. In the study group the mean (normalised) apoptotic keratocyte number was 1.1+/-1.7 in granular dystrophy and 0.5+/-1.1 in lattice type I dystrophy (p = 0.36, 0.63 respectively). Compared to the controls, the difference was statistically significant only for macular dystrophy (1.6+/-1.2; p = 0.01). Keratocyte apoptosis seems to be a concomitant or pathogenic factor in macular dystrophy. However, the pathways that are triggered to result in increased apoptotic cell death remain to be clarified.

Human sphingosine kinase: molecular cloning, functional characterization and tissue distribution.

Sphingosine-1-phosphate (SPP), the product of sphingosine kinase, is an important signaling molecule with intra- and extracellular functions. The cDNA for the mouse sphingosine kinase has recently been reported. In this paper we describe the cloning, expression and characterization of the human sphingosine kinase (huSPHK1). Sequence analysis comparison revealed that this kinase is evolutionarily very conserved, having a high degree of homology with the murine enzyme, and presenting several conserved regions with bacteria, yeast, plant, and mammalian proteins. Expressed huSPHK1 cDNA specifically phosphorylates D-erythro-sphingosine and, to a lesser extent, D, L-erythro-dihydrosphingosine, and not at all the 'threo' isoforms of dihydrosphingosine; hydroxy-ceramide or non-hydroxy-ceramide; diacylglycerol (DAG); phosphatidylinositol (PI); phosphatidylinositol-4-phosphate (PIP); or phosphatidylinositol-4, 5-bisphosphate (PIP(2)). huSPHK1 shows typical Michaelis-Menten kinetics (V(max)=56microM and K(m)=5microM). The kinase is inhibited by D,L-threo-dihydrosphingosine (K(i)=3microM), and by N, N-dimethyl-sphingosine (K(i)=5microM). Northern blots indicate highest expression in adult lung and spleen, followed by peripheral blood leukocyte, thymus and kidney, respectively. It is also expressed in brain and heart. In addition, database searches with the stSG2854 sequence indicate that huSPHK1 is also expressed in endothelial cells, retinal pigment epithelium, and senescent fibroblasts.

Signal-exon trap: a novel method for the identification of signal sequences from genomic DNA.

We describe a genomic DNA-based signal sequence trap method, signal-exon trap (SET), for the identification of genes encoding secreted and membrane-bound proteins. SET is based on the coupling of an exon trap to the translation of captured exons, which allows screening of the exon-encoded polypeptides for signal peptide function. Since most signal sequences are expected to be located in the 5'-terminal exons of genes, we first demonstrate that trapping of these exons is feasible. To test the applicability of SET for the screening of complex genomic DNA, we evaluated two critical features of the method. Specificity was assessed by the analysis of random genomic DNA and efficiency was demonstrated by screening a 425 kb YAC known to contain the genes of four secretory or membrane-bound proteins. All trapped clones contained a translation initiation signal followed by a hydrophobic stretch of amino acids representing either a known signal peptide, transmembrane domain or novel sequence. Our results suggest that SET is a potentially useful method for the isolation of signal sequence-containing genes and may find application in the discovery of novel members of known secretory gene clusters, as well as in other positional cloning approaches.

Expression of betaig-h3 is lower than normal in keratoconus corneas but increases with scarring.

PURPOSE: Keratoconus is a progressive ectatic disease of the cornea. Despite extensive clinical and laboratory investigations, its pathogenesis remains unclear. In this study, we examined the localization of betaig-h3, a recently described extracellular matrix protein in keratoconus corneas both in the absence and presence of subepithelial scarring. METHODS: Two normal corneas and central corneal buttons of 10 patients with keratoconus were excised during perforating keratoplasty and examined, including one case with acute corneal hydrops. In one case, keratoconus was associated with Down syndrome. Immunodetection was done with an antipeptide antibody reacting with the N-terminal part of betaig-h3. RESULTS: We found decreased betaig-h3 levels in the basal epithelial layer and keratocytes of keratoconus corneas. In the scarred corneas, however, betaig-h3 levels were increased in the basal epithelial layers and in activated keratocytes at the places of scarring. In the cornea of the patient with Down syndrome, we found an additional betaig-h3-positive zone in the anterior stroma. CONCLUSIONS: The decreased levels of betaig-h3 corneas seem to be specific for keratoconus. Considering the putative role of betaig-h3 as a cellular-attachment protein, paucity of betaig-h3 in the corneal stroma may lead to decreased mechanical stability and contribute to the development of keratoconus.

Dipeptidyl peptidase activity of CD26 in serum and urine as a marker of cholestasis: experimental and clinical evidence.

Dipeptidyl peptidase IV (CD26) is a membrane-associated enzyme that is expressed on the surface of T cells and on the hepatocyte brush border. In a soluble form it is present in serum. CD26 has been implicated in the regulation of T cell activation and in the metabolism of hormones and cytokines. Dipeptidyl peptidase (DPP) activity is elevated in the urine and serum of patients with biliary atresia (BA). To clarify the role of cholestasis in the development of increased serum and urinary DPP/CD26 activity, we studied the mechanism of activity increase in experimentally induced cholestasis of CD26-deficient and wild-type rats. The clinical utility of serum and urinary DPP/CD26 activity measurements was tested in adult and pediatric patients with hepatobiliary diseases and in liver transplant recipients. The results establish CD26-associated serum DPP activity as a novel, clinically useful marker of cholestasis and demonstrate that in contrast with alkaline phosphatase levels, DPP levels do not change in metastatic bone disease. Additionally, DPP activity is useful as a urinary test of cholestasis in infants who are not receiving nephrotoxic medication.

Immunohistochemical detection of betaIG-H3 in scarring human corneas.

BACKGROUND: BetaIG-H3 is a recently described extracellular matrix protein that is present in various organs. In rabbit corneas, increased betaIG (the rabbit form of betaIG-H3) mRNA levels were shown during corneal development and wound healing. In this study, we investigated the localization of betaIG-H3 protein in scarring human corneas. METHODS: Corneal buttons obtained during keratoplasty were examined. Immunohistological detection using a polyclonal antipeptide antibody against the betaIG-H3 protein was performed on 24 pathological corneas (9 ulcerations, 8 alkali burns, 2 perforating injuries, 5 bullous keratopathy) and 2 normal corneas. RESULTS: In normal corneas, strong staining was present in the basal layer of the epithelium and in the endothelium; the stromal fibers showed faint, uniform immunoreactivity. In all scarring corneas, the epithelium was usually thickened and all of its layers were reactive with the betaIG-H3 antibody. The cytoplasm of the stromal fibroblasts, as well as the stromal fibers around them also showed staining with the antibody. These changes were present in all scarring corneas, irrespective of the pathological process leading to scar formation. CONCLUSION: These results prove, at the protein level, the presence of increased amounts of betaIG-H3 at the sites of scarring in human corneas.

Corneal transplantation in children.

Authors discuss the special problems related to childhood keratoplasties. They evaluate the results of the keratoplasties carried out in children during the past 10 years in their service. They review the concerning literature and summarize their opinion on the corneal transplantations of this age group.

Characterization and chromosomal mapping of two pseudogenes of the mouse Pafaha/Lis1 gene: retrointegration hotspots in the mouse genome.

Isolated lissencephaly sequence and Miller-Dieker syndrome are related neurodevelopmental disorders caused by defects of the LIS1 gene encoding the alpha subunit of intracellular platelet-activating factor acetylhydrolase. In addition to the ortholog of the human LIS1 gene (Pafaha/Lis1), the mouse genome contains two more homologs. In order to characterize the new members of this gene family, we isolated both Pafaha/Lis1-related genes (Pafaha-ps1 and Pafaha-ps2) from a mouse genomic library. Pafaha-ps1 and Pafaha-ps2 are processed pseudogenes formed by the retroinsertion of 5'-truncated Pafaha/Lis1 cDNAs. Sequence analysis revealed a striking accumulation of retroelements at both loci, identifying two retroinsertion hotspots in the mouse genome. The recognition of tRNA genes flanking Pafaha-ps1 provides an example for the potential association of RNA polymerase III transcription and retroinsertion in mammals. Linkage mapping placed Pafaha-ps1 and Pafaha-ps2 to distal chromosome (Chr) 3 and proximal Chr 7, respectively. Our results indicate that only one of the three LIS1-related mouse loci (Pafaha/Lis1) is functional, in contrast with two closely related functional genes (LIS1 and LIS2) reported in humans. 1998 Elsevier Science B.V.