Establishment of killer whale (Orcinus orca) primary fibroblast cell cultures and their transcriptomic responses to pollutant exposure.

Populations of killer whale (Orcinus orca) contain some of the most polluted animals on Earth. Yet, the knowledge on effects of chemical pollutants is limited in this species. Cell cultures and in vitro exposure experiments are pertinent tools to study effects of pollutants in free-ranging marine mammals. To investigate transcriptional responses to pollutants in killer whale cells, we collected skin biopsies of killer whales from the Northern Norwegian fjords and successfully established primary fibroblast cell cultures from the dermis of 4 out of 5 of them. Cells from the individual with the highest cell yield were exposed to three different concentrations of a mixture of persistent organic pollutants (POPs) that reflects the composition of the 10 most abundant POPs found in Norwegian killer whales (p,p'-DDE, trans-nonachlor, PCB52, 99, 101, 118, 138, 153, 180, 187). Transcriptional responses of 13 selected target genes were studied using digital droplet PCR, and whole transcriptome responses were investigated utilizing RNA sequencing. Among the target genes analysed, CYP1A1 was significantly downregulated in the cells exposed to medium (11.6 µM) and high (116 µM) concentrations of the pollutant mixture, while seven genes involved in endocrine functions showed a non-significant tendency to be upregulated at the highest exposure concentration. Bioinformatic analyses of RNA-seq data indicated that 13 and 43 genes were differentially expressed in the cells exposed to low and high concentrations of the mixture, respectively, in comparison to solvent control. Subsequent pathway and functional analyses of the differentially expressed genes indicated that the enriched pathways were mainly related to lipid metabolism, myogenesis and glucocorticoid receptor regulation. The current study results support previous correlative studies and provide cause-effect relationships, which is highly relevant for chemical and environmental management.

Persistent organic pollutants and organophosphate esters in feathers and blood plasma of adult kittiwakes (Rissa tridactyla) from Svalbard - associations with body condition and thyroid hormones.

Polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs), organochlorine pesticides (OCPs) and organophosphate esters (OPEs) were assessed in blood plasma and feathers of 19 adult black-legged kittiwakes (Rissa tridactyla) breeding in two colonies (Blomstrandhalvøya and Krykkjefjellet) at the Arctic archipelago, Svalbard. Potential associations with body condition index (BCI) and thyroid hormones were investigated. All compound classes were detected in both blood plasma and feathers, but due to low sample size and volumes, OPEs could only be quantified in four individuals, warranting larger follow-up studies. Kittiwakes breeding at Blomstrandhalvøya had significantly higher concentrations of organic pollutants in blood plasma than kittiwakes breeding at Krykkjefjellet (p < 0.001). Concentrations in blood plasma and feathers did not significantly correlate for any of the investigated compounds, and feather concentrations did not differ significantly between the colonies. This suggests that pollutant levels in adult kittiwake feathers do not reflect local contamination at breeding sites and are as such not useful to monitor local contamination at Svalbard. Significant negative associations between BCI and most pollutants were found in both populations, whereas significant correlations between the BCI, the ratio of total triiodothyronine to free triiodothyronine (TT3:fT3), and several pollutants were only found for kittiwakes from Blomstrandhalvøya (all r ≥ -0.60 and p ≤ 0.05). This indicates that higher levels of circulating pollutants during the breeding period covary with the TT3: fT3 ratio, and may act as an additional stressor during this period.

Towards an alternative testing strategy for nanomaterials used in nanomedicine: lessons from NanoTEST.

In spite of recent advances in describing the health outcomes of exposure to nanoparticles (NPs), it still remains unclear how exactly NPs interact with their cellular targets. Size, surface, mass, geometry, and composition may all play a beneficial role as well as causing toxicity. Concerns of scientists, politicians and the public about potential health hazards associated with NPs need to be answered. With the variety of exposure routes available, there is potential for NPs to reach every organ in the body but we know little about the impact this might have. The main objective of the FP7 NanoTEST project ( www.nanotest-fp7.eu ) was a better understanding of mechanisms of interactions of NPs employed in nanomedicine with cells, tissues and organs and to address critical issues relating to toxicity testing especially with respect to alternatives to tests on animals. Here we describe an approach towards alternative testing strategies for hazard and risk assessment of nanomaterials, highlighting the adaptation of standard methods demanded by the special physicochemical features of nanomaterials and bioavailability studies. The work has assessed a broad range of toxicity tests, cell models and NP types and concentrations taking into account the inherent impact of NP properties and the effects of changes in experimental conditions using well-characterized NPs. The results of the studies have been used to generate recommendations for a suitable and robust testing strategy which can be applied to new medical NPs as they are developed.

Umbilical cord length in singleton gestations: a Finnish population-based retrospective register study.

INTRODUCTION: Many complications of pregnancy and delivery are associated with umbilical cord length. It is important to examine the variation in length, in order to identify normal and abnormal conditions. Moreover, the factors influencing cord growth and development are not precisely known. OBJECTIVE: The main objectives were to provide updated reference charts for umbilical cord length in singleton pregnancies and to evaluate potential factors affecting cord length. METHODS: Birth register data of 47,284 singleton pregnant women delivering in Kuopio University Hospital, Finland was collected prospectively. Gender-specific centile charts for cord length from 22 to 44 gestational weeks were obtained using generalized additive models for location, scale, and shape (GAMLSS). Gestational, fetal, and maternal factors were studied for their potential influence on cord length with single variable analysis and stepwise multiple linear regression analysis. RESULTS: Cord length increased according to gestational age, while the growth decelerated post-term. Birth weight, placental weight, pregravid maternal body mass index, parity, and maternal age correlated to cord length. Gestational diabetes and previous miscarriages were associated with longer cords, while female gender and placental abruption were associated with shorter cords. DISCUSSION AND CONCLUSIONS: Girls had shorter cords throughout gestation although there was substantial variation in length in both genders. Cord length associated significantly with birth weight, placental weight, and gestational age. Significantly shorter cords were found in women with placental abruption. This important finding requires further investigation.

Reduction of intra-ocular pressure by ocular compression before combined peri- and retrobulbar block.

The intra-ocular pressure immediately before glaucoma surgery can be raised. We wished to investigate if ocular compression for 20 min before a combined peri- and retrobulbar injection would result in a lower pressure after the block. Sixty consecutive patients scheduled for filtration surgery were randomly assigned to receive ocular compression using an external pressure device for 20 min before combined peri- and retrobulbar injection (intervention group, who also received compression after the block) or to a control group in whom pressure was applied only after the block was completed. The intra-ocular pressure was measured at baseline, after the 20-min pre-injection compression (intervention group), after injecting the block, and after the 10-min post-injection compression. The pressure did not differ between groups at baseline, after the block or after the post-injection compression. In the intervention group, the compression before the block reduced the median (IQR [range]) pressure from 21.0 (17.0-25.0 [12.0-40.0]) mmHg to 16.8 (12.5-22.5 [7.5-33.5]) mmHg (p<0.001). We conclude that external ocular compression reduces the intra-ocular pressure, but applying an additional compression for 20 min before injecting the block is not beneficial.

Vacuum-assisted delivery is associated with late-onset asthma.

BACKGROUND: Perinatal factors during delivery might modulate fetal immunological development and thereby be associated with the development of allergic diseases and asthma later. METHODS: Perinatal data was recorded during pregnancy and at the time of delivery in regard to 5823 children who were born in Northern Finland in 1985-1986. Data from self-administered questionnaires were available at the ages of 7 and 15-16 years and skin prick tests for four main allergens were carried out at the age of 15-16 years. Only singletons delivered by the vaginal route were analyzed. RESULTS: There was a higher prevalence of doctor-diagnosed asthma at any time of life among children who were delivered by vacuum extraction (RR 1.80, 95% CI 1.27-2.56; P < 0.001) in comparison with spontaneously delivered children. In particular, this risk was increased as regards late-onset asthma (RR 2.41, 95% CI 1.52-3.81; P < 0.001). Perinatal effects had less impact on the development of other asthma, atopy or hay fever. CONCLUSIONS: The delivery by vacuum extraction had significant impact on the development of late-onset asthma compared with spontaneously delivered children.

Reversal of optic disc cupping in glaucoma.

AIM: To study whether reversal of optic disc cupping after intraocular pressure (IOP) reduction is related to risk of glaucoma progression. METHODS: In this prospective follow-up study, where 51 patients with exfoliation glaucoma and five with ocular hypertension combined with exfoliation syndrome were followed for 6 years after IOP reduction, 24 showed progression of glaucoma in visual fields or optic nerve head (ONH) stereophotographs. ONH topography was measured with the Heidelberg Retina Tomograph (HRT). A decrease in HRT parameter cup volume of more than 5% was considered cup reversal. Multiple logistic regression was used to model progression of glaucoma. RESULTS: Cup reversal (OR 0.226; 95% CI 0.055 to 0.918, p = 0.037), final IOP (OR 1.216; 95% CI 1.000 to 1.479, p = 0.050) and visual field mean defect at entry (OR 1.158; 95% CI 1.034 to 1.296, p = 0.011) were associated with progression. IOP change from study entry to 6-year control visit was not associated with progression (OR 0.964, 95% CI 0.850 to 1.092, p = 0.56). CONCLUSION: Cup reversal seemed to be an independent protective factor for progression of glaucoma.

Individual patient variation and inter-rater reliability of lower calyceal infundibular width on routine intravenous pyelography.

OBJECTIVE: To standardise infundibular width (IFW) measurement, to determine patient variability, and to determine inter-rater variability on intravenous pyelography (IVP). PATIENTS AND METHODS: Fifty outpatient IVP films were randomly drawn from two hospitals between 1 July 1998 and 31 August 1999, and examined to measure the lower calyceal IFW on the 5, 10 and 20 min films with compression, and after voiding. Kidneys with previous renal surgery, hydronephrosis or renal anatomical anomalies were excluded; in all, 81 kidneys from 50 patients were examined. The IFW was measured at the narrowest point along the infundibulum. All 50 films were then reviewed by two urologists unaware of their origin, to determine the inter-rater reliability of the infundibular measurements. RESULTS: Analysis of variance (anova) with posthoc analysis showed a significant difference in IFW at each phase of the IVP (repeated measures anova, P < 0.001). The mean (95% confidence interval) IFW was greatest on the compression film, at 4.4 (2.6) mm, and least on the postvoid film, at 1.6 (2.1) mm. Overall inter-rater reliability was 0.9780 (intraclass correlation coefficient), and the Pearson correlation between each rater for the IFW at each phase of the IVP was >/= 0.886. CONCLUSIONS: There is wide variability in lower calyceal IFW among the various IVP films in a given study. Thus any predictive value of the IFW must be standardized for the timing and IVP film type (compression, postvoid, etc.). The inter-rater reliability of IFW is high, suggesting that if used correctly it may be more useful in predicting the outcome after shock wave lithotripsy.

Blood flow of the optic nerve head and peripapillary retina in exfoliation syndrome with unilateral glaucoma or ocular hypertension.

BACKGROUND: The purpose was to study whether any differences exist in the optic nerve head (ONH) and peripapillary retinal blood flow between the two eyes of patients with unilateral exfoliation glaucoma or ocular hypertension (OHT) with exfoliation syndrome. METHODS: This cross-sectional study included 50 patients. All had exfoliation syndrome with glaucoma or OHT in one eye, and these eyes comprised the study group. The fellow eyes, all normotensive, comprised the control group. Blood flow was measured with scanning laser Doppler flowmetry in the lamina cribrosa region, in the rim area, and on the peripapillary retina. Multiple linear regression analyses were used to identify any associations between different factors and differences in flow. RESULTS: Flow in the rim area was significantly higher in the study eyes than in the control eyes, with a mean difference of 172 arbitrary units (P = 0.001). The difference of 40 units in the laminar area was of borderline significance (P = 0.065) and no significant difference was found in the peripapillary retina (P = 0.530). In the study eyes, blood flow of the ONH lamina and rim area decreased with increasing glaucomatous damage, and treatment with topical timolol was associated with reduced blood flow in the lamina cribrosa and rim area. Perfusion pressure was associated only with flow in the peripapillary area (P = 0.021). CONCLUSIONS: Advanced glaucomatous damage was associated with reduced flow both in the lamina cribrosa and the rim area but not in the peripapillary retina. Treatment with topical timolol was associated with decreased flow in the ONH.

Scanning laser ophthalmoscopy of the optic nerve head in exfoliation glaucoma and ocular hypertension with exfoliation syndrome.

AIMS: To study the relation between optic nerve head topography (Heidelberg retina tomograph, HRT) and disc area, visual field index mean defect (MD), and intraocular pressure (IOP), and to see whether change in HRT parameters is associated with change in MD in a prospective follow up. METHODS: 80 consecutive patients (69 patients with exfoliation glaucoma and 11 with ocular hypertension combined with exfoliation syndrome) were examined before IOP reducing intervention and prospectively followed every 6 months for 2 years. RESULTS: At the entry point, multiple regression analysis showed significant linear association between MD and all HRT parameters, when controlling for disc area. Disc area showed significant association with cup area, cup/disc area ratio, rim area, cup volume, and mean RNFL thickness. Six months after intervention IOP had decreased significantly. Reversible changes in cup area, cup/disc area ratio, rim area, cup volume, rim volume, mean cup depth, and maximum cup depth were associated with decrease in IOP. During the follow up period from 6 month to 2 years, IOP did not change significantly, and MD was used as a measure of progression of glaucoma. During this period, only cup shape measure among HRT parameters showed significant association with subsequent change in MD. CONCLUSION: Disc area should be taken into account when using HRT to compare patients. Any effect of change in IOP should be also taken into account when using HRT in follow up. Cup shape measure is a promising indicator of progression of glaucomatous damage.

Corneal thickness and corneal endothelium in normotensive subjects with unilateral exfoliation syndrome.

PURPOSE: To examine the central corneal thickness (CCT) and corneal endothelium in both eyes of patients with unilateral exfoliation syndrome (EXS). To determine the effect of CCT on the measurement of intraocular pressure (IOP). METHODS: In this cross-sectional clinical study, comparisons were made of CCT (Humphrey Ultrasonic Pachometer), corneal endothelial cells (Keeler-Konan contact specular microscope) and IOP (Goldmann applanation tonometer) between the exfoliative (E) and fellow nonexfoliative (NE) eyes in 40 normotensive patients with unilateral EXS. The CCT was used to obtain a corrected value for the IOP. RESULTS: The E eyes had significantly higher values for CCT (0.528+/-0.030 vs 0.523+/-0.032 mm, P<0.01) and IOP (15.7+/-3.6 vs 14.4+/-2.9 mmHg, P<0.001) than the fellow NE eyes. The paired E and NE eyes did not differ in endothelial cell density (2779+/-540 vs 2870+/-386 cells/mm2), in the coefficient of variation of cell size (0.25+/-0.03 vs 0.26+/-0.03) or in the frequency of hexagonal cells (80.5+/-6.5 vs 82.0+/-5.0%). After correcting IOP for CCT, the E eyes still had significantly higher IOP than the NE eyes (15.1+/-4.4 vs 14.2+/-3.7 mmHg, P<0.05). CONCLUSION: Normotensive eyes with early EXS did not have quantitative (cell density) or qualitative (variation in cell size, frequency of hexagonal cells) morphological changes in corneal endothelium, but had higher values for IOP and CCT. After correcting IOP for CCT, the E eyes still had significantly higher IOP than the fellow NE eyes.

Intraocular pressure and progression in exfoliative eyes with ocular hypertension or glaucoma.

PURPOSE: This retrospective study was conducted in exfoliative eyes in order to study the influence of IOP and other possible factors on the conversion from ocular hypertension (OHT) to glaucoma and progression of glaucoma. METHODS: 139 patients with exfoliation OHT or glaucoma with exfoliation syndrome were included (mean follow-up 5.2 +/- 3.6 years). The effects of age, gender, weighted mean IOP, maximum IOP, stage of glaucoma at the first visit, refraction, glaucoma medication, and interventions on the hazard of progression were studied by multivariate survival analysis. RESULTS: Conversion from OHT to glaucoma or progression of glaucoma was detected in 63 eyes (45.3%). A significant association with progression was found for age (relative risk 1.042; p-value 0.043), weighted mean IOP (1.076; <0.001), and stage of glaucoma (1.436; <0.001). History of trabeculectomy (0.360; 0.002) related to a decreased risk. CONCLUSION: The risk of conversion from OHT to glaucoma and progression of glaucoma increased in exfoliative eyes with severity of stage of glaucoma, weighted mean IOP, and the age of the patient. A history of trabeculectomy was related to decreased risk of progression.

Optic disc topography before and after trabeculectomy in advanced glaucoma.

BACKGROUND AND OBJECTIVE: To analyze changes in the optic disc topography after filtration surgery. PATIENTS AND METHODS: Laser scanning tomography was performed in 10 eyes of 9 patients (mean age 65.6 +/- 8.1 years; age range 55 to 75 years) after a mean follow-up of 3.7 months, and in 8 eyes of 7 patients (mean age 63.1 +/- 7.6 years; age range 55 to 75 years) after a mean follow-up of 13.1 months. RESULTS: Preoperatively, the mean intraocular pressure (IOP) was 24.4 +/- 6.9 mm Hg with maximal medication. Postoperatively at 3.7 months it was 11.0 +/- 2.2 mm Hg, and at l2.1 months it was l3.6 +/- 2.8 mm Hg. An initial reduction in IOP of more than 30 percent at 3.7 months was achieved without medication in all but 1 eye. After 12.1 months, an IOP reduction of more than 30 percent was achieved in 6 of 8 eyes (1 with medication), while 2 eyes had IOP reductions of less than 15 percent (< or = 3 mm Hg) despite medication. In the 6 eyes, the optic disc cup volume showed a decrease of more than 30 percent, while the 2 eyes without marked IOP reductions had slight increases in cup volume. Of the 2 eyes without marked decreases in cup size, 1 had normal-tension glaucoma and 1 experienced an increase in IOP of more than 25 mm Hg during the first postoperative week. CONCLUSION: The postoperative IOP should be kept low enough to permit reversal of optic disc changes.

Postpartum recovery after severe pre-eclampsia and HELLP-syndrome.

Postpartum recovery was examined in 100 pregnancies complicated by severe pre-eclampsia, and in 15 pregnancies in which HELLP-syndrome as present. Albuminuria disappeared and diastolic blood pressure returned to normal (< 90 mmHg) in half of the cases within one week postpartum. Postpartum recovery in the cases with HELLP-syndrome did not differ from that seen in the pre-eclamptic patients. Thrombocytopenia showed spontaneous resolution within three days after delivery. In stepwise discriminant analysis, the incidence of IUGR predicted a slow postpartum recovery, but this did not apply to other factors. Such as diastolic blood pressure before delivery, duration of subjective symptoms, duration of pregnancy or severity of albuminuria. Elevated diastolic pressure and/or albuminuria were diagnosed two months postpartum in one third of the patients. If the development of eclampsia is prevented by correctly timing the induction of delivery, relatively good short-term postpartum recovery is probable after severe pre-eclampsia and HELLP-syndrome.

A randomized, prospective study on the use of sodium hyaluronate (Healon) in trabeculectomy.

The effect of injecting sodium hyaluronate (Healon) before fistulization in trabeculectomy was investigated in a prospective, randomized study of 107 consecutive eyes. Fifty-two eyes were operated on with the viscoelastic and 55 without. An early postoperative intraocular pressure (IOP) peak of 21 mm Hg or more occurred in 27% of the study eyes versus 17% of the control eyes, an insignificant difference. The mean peak IOPs of the study and the control eyes did not differ significantly (29.8 +/- 9.0 mm Hg versus 32.7 +/- 10.2 mm Hg), nor did the mean IOPs and the need for glaucoma medications 1 month after surgery. Shallow/flat anterior chamber and choroidal detachment were equally frequent in both groups, but hyphema occurred significantly more often in the study group (40%) than in the control group (18%).

Comparative effects of alpha-2 receptor agents and THA on the performance of adult and aged rats in the delayed non-matching to position task.

The present study investigated the effects of dexmedetomidine (an alpha-2 adrenoceptor agonist), atipamezole (an alpha-2 adrenoceptor antagonist) and tacrine (an inhibitor of acetylcholinesterase) on the performance of adult and aged rats in a delayed non-matching to position task assessing spatial short-term memory. Most of the aged rats were impaired in the pretraining phases and in the acquisition of the non-delayed version of the task. After a substantial training period of the delayed version of the task, both adult and aged rats reached their asymptotic level of performance. Both adult and aged rats showed a decline in the percent correct responses at the longest delays in this task, and a delay independent decrease in the percent correct responses across the delays (0-30 s) was found in the group of aged rats (25-month-old) as compared to the adults (10-month-old). Dexmedetomidine (0.3, 1.0 or 3.0 micrograms/kg), atipamezole (0.03, 0.3 or 3.0 mg/kg) and tacrine (1.0 or 3.0 mg/kg) did not increase the percent correct responses in adult or aged rats. The highest doses of dexmedetomidine and tacrine decreased behavioural activity of rats during this short-term memory testing. Atipamezole (0.03 mg/kg) increased behavioural activity of rats. The results suggest that acute, systemic administrations of alpha-2 drugs or an anticholinesterase do not improve short-term memory in rats.

Occlusal anomalies in 45,X/46,XX- and 46,Xi(Xq)-women (Turner syndrome).

A total of 20 individuals with X-chromosome aberrations, 14 45,X/46,XX-women and six 46,Xi(Xq)-women, were examined for occlusal anomalies. The controls were first-degree woman relatives and population women. The results showed that the most common type of malocclusion in both study groups was lateral cross bite. Also distal molar occlusion, increased maxillary overjet and in 45,X/46,XX-women open bite was found. 45,X/46,XX-women's occlusion seems to be more affected than that of 46,Xi(Xq)-women. Compared to 45,X-women, the present study groups show milder expression of malocclusion. One might speculate that enamel genes of the X-chromosome are involved in occlusal development.