Contacting co-culture of human retinal microvascular endothelial cells alters barrier function of human embryonic stem cell derived retinal pigment epithelial cells.

Here we evaluated the effects of human retinal microvascular endothelial cells (hREC) on mature human embryonic stem cell (hESC) derived retinal pigment epithelial (RPE) cells. The hESC-RPE cells (Regea08/017, Regea08/023 or Regea11/013) and hREC (ACBRI 181) were co-cultured on opposite sides of transparent membranes for up to six weeks. Thereafter barrier function, small molecule permeability, localization of RPE and endothelial cell marker proteins, cellular fine structure, and growth factor secretion of were evaluated. After co-culture, the RPE specific CRALBP and endothelial cell specific von Willebrand factor were appropriately localized. In addition, the general morphology, pigmentation, and fine structure of hESC-RPE cells were unaffected. Co-culture increased the barrier function of hESC-RPE cells, detected both with TEER measurements and cumulative permeability of FD4 - although the differences varied among the cell lines. Co-culturing significantly altered VEGF and PEDF secretion, but again the differences were cell line specific. The results of this study showed that co-culture with hREC affects hESC-RPE functionality. In addition, co-culture revealed drastic cell line specific differences, most notably in growth factor secretion. This model has the potential to be used as an in vitro outer blood-retinal barrier model for drug permeability testing.

[A phd completed. The effect of radiotherapy on oral mucosa cell morphology.] / Serie: Hora est. Het effect van radiotherapie op de morfologie van de orale mucosa.

The treatment of oral cancer usually consists of surgical removal of the tumour, possibly followed by radiotherapy. The purpose of this dissertation research was to investigate the effects of radiotherapy on the oral tissues, in particular the superficially positioned epithelial cells in the oral mucosa. Earlier studies with electron microscopy revealed that unradiated oral mucosa cells, when observed at high magnification, contain microplicae (ridges or folds). Together with various saliva components, these microplicae form a protective layer that offers defence against, for instance, microorganisms. Radiotherapy damages these microplicae and may even destroy them altogether. Studies have shown that this effect of radiation can be observed in animals as well as in humans. As the radiation dose increased (50 Gy or more) the destruction of the microplicae was more severe. With a dose of 60 Gy or more the microplicae completely disappeared. This process may play an important role in the occurrence of osteoradionecrosis in the jaw and failure of dental implants placed after radiotherapy.

Synergistic proinflammatory interactions of microbial toxins and structural components characteristic to moisture-damaged buildings.

Indoor exposure to microbes and their structural and metabolic compounds is notoriously complex. To study proinflammatory interactions between the multiple microbial agents, macrophages derived from human THP-1 monocytic cells were exposed to several concentrations of microbial toxins alone (emodin, enniatin B, physcion, sterigmatocystin, valinomycin) and in combination with microbial structural components (bacterial lipopolysaccharide [LPS] or fungal ß-glucan). While the expression of proinflammatory cytokines TNFα and IL-1ß to single toxins alone was modest, low-dose co-exposure with structural components increased the responses of emodin, enniatin B, and valinomycin synergistically, both at the mRNA and protein level, as measured by RT-qPCR and ELISA, respectively. Co-exposure of toxins and ß-glucan resulted in consistent synergistically increased expression of several inflammation-related genes, while some of the responses with LPS were also inhibitory. Co-exposure of toxins with either ß-glucan or LPS induced also mitochondrial damage and autophagocytosis. The results demonstrate that microbial toxins together with bacterial and fungal structural components characteristic to moisture-damaged buildings can have drastic synergistic proinflammatory interactions at low exposure levels.

Microplicae--Specialized Surface Structure of Epithelial Cells of Wet-Surfaced Oral Mucosa.

The surface structure of the superficial cells of the oral mucosa is decorated with numerous membrane ridges, termed microplicae (MPLs). The MPL structure is typical of the epithelial surfaces that are covered with protective mucus. Cell membrane MPLs are no longer seen as passive consequences of cellular activity. The interaction between MPLs and the mucins has been demonstrated, however the role of MPL structure seen on the upper surface of the oral epithelial cells is speculative. The cell surface is of potentially great significance, as it harbors many markers for refined prognosis and targets for oral mucosal diseases and cancer therapy. With these aspects in mind, we conducted the present review of the MPL structure and function in order to form the basis for further studies of MPLs of the oral epithelial cells.

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.

Analysis of plastic deformation in cortical bone after insertion of coated and non-coated self-tapping orthopaedic screws.

Insertion of internal fracture fixation devices, such as screws, mechanically weakens the bone. Diamond-like carbon has outstanding tribology properties which may decrease the amount of damage in tissue. The purpose of this study was to investigate methods for quantification of cortical bone damage after orthopaedic bone screw insertion and to evaluate the effect of surface modification on tissue damage. In total, 48 stainless steel screws were inserted into cadaver bones. Half of the screws were coated with a smooth amorphous diamond coating. Geometrical data of the bones was determined by peripheral quantitative computed tomography. Thin sections of the bone samples were prepared after screw insertion, and histomorphometric evaluation of damage was performed on images obtained using light microscopy. Micro-computed tomography and scanning electron microscopy were also used to examine tissue damage. A positive correlation was found between tissue damage and the geometric properties of the bone. The age of the cadaver significantly affected the bone mineral density, as well as the damage perimeter and diameter of the screw hole. However, the expected positive effect of surface modification was probably obscured by large variations in the results and, thus, statistically significant differences were not found in this study. This can be explained by natural variability in bone tissue, which also made automated image analysis difficult.

Tissue viscoelasticity is related to tissue composition but may not fully predict the apparent-level viscoelasticity in human trabecular bone - An experimental and finite element study.

Trabecular bone is viscoelastic under dynamic loading. However, it is unclear how tissue viscoelasticity controls viscoelasticity at the apparent-level. In this study, viscoelasticity of cylindrical human trabecular bone samples (n=11, male, age 18-78 years) from 11 proximal femurs were characterized using dynamic and stress-relaxation testing at the apparent-level and with creep nanoindentation at the tissue-level. In addition, bone tissue elasticity was determined using scanning acoustic microscope (SAM). Tissue composition and collagen crosslinks were assessed using Raman micro-spectroscopy and high performance liquid chromatography (HPLC), respectively. Values of material parameters were obtained from finite element (FE) models by optimizing tissue-level creep and apparent-level stress-relaxation to experimental nanoindentation and unconfined compression testing values, respectively, utilizing the second order Prony series to depict viscoelasticity. FE simulations showed that tissue-level equilibrium elastic modulus (Eeq) increased with increasing crystallinity (r=0.730, p=.011) while at the apparent-level it increased with increasing hydroxylysyl pyridinoline content (r=0.718, p=.019). In addition, the normalized shear modulus g1 (r=-0.780, p=.005) decreased with increasing collagen ratio (amide III/CH2) at the tissue-level, but increased (r=0.696, p=.025) with increasing collagen ratio at the apparent-level. No significant relations were found between the measured or simulated viscoelastic parameters at the tissue- and apparent-levels nor were the parameters related to tissue elasticity determined with SAM. However, only Eeq, g2 and relaxation time τ1 from simulated viscoelastic values were statistically different between tissue- and apparent-levels (p<.01). These findings indicate that bone tissue viscoelasticity is affected by tissue composition but may not fully predict the macroscale viscoelasticity in human trabecular bone.

Long term treatment of severe hypercholesterolaemia with guar gum.

The long term efficacy of granulated guar gum, 15-30 g per day, was studied in 23 patients with severe hypercholesterolaemia (serum cholesterol concentration between 8.0 and 14.3 mmol/l). Originally, 29 patients participated in the study. Two patients dropped out because of gastrointestinal side effects, two others were not willing to complete the study without any given reason, and two discontinued the study because of hospitalization. A 1-month placebo period preceded the guar gum treatment, and another 1-month placebo period followed after 50 weeks of active treatment. The serum total cholesterol concentration (mean +/- SEM) was reduced from 10.0 +/- 0.4 mmol/l to 8.2 +/- 0.3 mmol/l (P less than 0.001) after 8 weeks and to 9.0 +/- 0.4 mmol/l (P less than 0.001) after 50 weeks on guar gum. During the second placebo period serum cholesterol returned to the pretreatment level. After 34 weeks of active treatment the serum LDL-cholesterol concentration had fallen by 15% and that of apoprotein B by 14% from the baseline. The changes in lipid and lipoprotein levels were independent of the initial values and the type of hypercholesterolaemia. Serum triglycerides, HDL-cholesterol, body weight and blood pressure showed no significant changes during the trial. Of the study subjects, 20 reached the maximum intended dose of 30 g per day guar gum between 8 and 14 weeks and thereafter 11 subjects continued the dose of 30 g/day while 12 subjects reduced the dose to 15-25 g/day.(ABSTRACT TRUNCATED AT 250 WORDS)

Low diastolic blood pressure and mortality in a population-based cohort of 16913 hypertensive patients in North Karelia, Finland.

OBJECTIVE: To examine the relationship between mortality and diastolic blood pressure during treatment. DESIGN: A prospective follow-up of a population-based dynamic cohort of hypertensive patients. SETTING: Province of North Karelia, eastern Finland. PATIENTS: A cohort of 16 913 North Karelian hypertensive patients in hypertension register of the North Karelia Project, who had been followed up since 1972 until the end of 1985. Most of these patients had been under antihypertensive drug therapy during the follow-up. MAIN OUTCOME MEASURES: Death: all deaths (n = 4490), deaths from cardiovascular disease (n = 2995) and deaths from non-cardiovascular disease (n = 1495). RESULTS: Of all deaths, 17% of those among men and 24% of those among women were of patients with mean diastolic blood pressures less than 90 mmHg. We found a U-shaped relationship between diastolic blood pressure and total, cardiovascular and non-cardiovascular mortalities. We investigated this relationship in more detail using Cox regression model. Low diastolic blood pressure was associated independently with increased mortality for the patients aged 50 years or more at baseline. The occurrence of cardiac failure and other cardiovascular complications of hypertension were more important determinants of mortality than was low diastolic blood pressure alone. CONCLUSIONS: We demonstrated that there is an association between low diastolic blood pressure and mortality for treated hypertensive patients aged 50-69 years. The clinical importance of this relationship for patients without any cardiovascular complications of hypertension seems negligible. For patients with complications, these complications are likely to be primary factors causing greater than normal mortality and low diastolic blood pressure is mostly a secondary phenomenon. Our data do not lend support to speculations that there is overtreatment of hypertension, which would increase mortality through making diastolic blood pressures too low.

Antidromic corticospinal tract potential of the brain.

OBJECTIVE: To describe a novel potential component (antidromic corticospinal tract potential, ACSP) of the brain after translaminar spinal stimulation of a relaxed patient during scoliosis surgery. To study the origin of this component and to compare its source to known sources of the somatosensory evoked potentials (SEPs). METHODS: We studied 17 consecutive patients during posterior scoliosis surgery. SEPs and ACSPs were elicited by translaminar spinal stimulation at the Th 2 and L 1 levels. ACSPs and SEPs were recorded on the scalp midline. Neurogenic motor evoked potentials (NMEPs) were recorded on the popliteal spaces. Preoperative tibial SEPs were also recorded. RESULTS: ACSP was distinctly separated from the corresponding spinally evoked cortical SEP that showed longer latency than the ACSP. ACSPs decreased and disappeared when stimulation was moved to the caudal direction in the conus region while SEP persisted. In addition, the hemispheric origin of ACSP was confirmed with multichannel midline recordings of the scalp and neck. Thus there was no confusion to the response of nucleus gracilis, corresponding the P 31 response of the tibial nerve SEP. CONCLUSIONS: The origin of ACSP seemed to be in the rostral part of the corticospinal tract. ACSP diminished in the conus region when stimulation was moved caudally and it disappeared when the stimulus was given to the root level. This proves that ACSP is not a response of the somatosensory tract, instead ACSP represents antidromic response of the pyramidal tract. ACSP can be used in monitoring of the motor tracts during scoliosis surgery together with NMEPs.

Subspace regularization method for the single-trial estimation of evoked potentials.

A method for the single-trial estimation of the evoked potentials is proposed. The method is based on the so-called subspace regularization approach in which the second-order statistics of the set of the measurements is used to form a prior information model for the evoked potentials. The method is closely related to the Bayesian estimation. The performance of the proposed method is evaluated using realistic simulations. As a specific application the method is applied to the estimation of the target responses in the P300 test.

Analysis of galvanic skin responses with principal components and clustering techniques.

An advanced method for analyzing the patterning of successive galvanic skin responses (GSR) is presented. The proposed method is based on principal component analysis in which the vector containing the measured signal is presented as a weighted sum of orthogonal basis vectors. The method is tested using measurements from 20 healthy controls and 13 psychotic patients. For each subject, 11 surprising auditory stimuli were delivered to right ear at irregular intervals and evoked GSRs were recorded from the hand. For most of the healthy controls, there was a clear pattern in successive GSRs, whereas within psychotic patients the lack of time-locking of GSRs seemed to be characteristical. These between group differences can be revealed by the proposed method. With application to clustering a significant discrimination, with overall correct ratings of 82%, of healthy controls and psychotic patients is achieved. A significant fact is that all patients were ranked correctly giving the proposed method a sensitivity of 100%.

Single-trial estimation of multichannel evoked-potential measurements.

A method for single-trial estimation of multichannel evoked potentials is presented. The proposed method is based on the regularized least squares scheme. The spatial correlation between the channels is used as additional information in the estimation procedure. Amplitude estimates obtained with the proposed method is compared with the estimates calculated without using the spatial information. The performance of the method is evaluated using simulated and real data of P300 responses measured using auditory stimuli. The multichannel approach is shown to give realistic and comparable information about the amplitude differences of the P300 peak between different channels.

The Edinburgh-Helsinki study: a comparison of dental care for children.

A comparison was undertaken of dental care for children in Edinburgh and Helsinki. The evaluation centred upon an epidemiological investigation of the dental health of children in the 5-, 12- and 15-year age groups. This was performed by examiners using standardized procedures who were drawn from both countries. They employed an established system of data recording and processing (SPEED). Using these data, and background information from official and other sources, an assessment was made of the effectiveness, adequacy, efficiency and appropriateness of the services. Children's dental health, particularly in the youngest age group, was generally better in Helsinki than in Edinburgh as were both the population coverage and extent to which disease was controlled. However, the services in Edinburgh were marginally the more economically efficient. This arose from their being provided mainly by independent contractors working under a fee per item of service rather than by a public salaried system. In neither city was the delivery of dental services supplemented by water fluoridation nor was extensive use made of auxiliary personnel. The decrease in caries prevalence in recent years has profound implications for the dental services in both countries. It calls for continuing re-appraisal of present policies on expenditure, manpower, dental education and ways of delivering dental care.

Apparatus to test insertion and removal torque of bone screws.

This paper describes affordable equipment for testing bone screw torque, corresponding to ASTM standard F543-00 for testing metallic medical bone screws. Correct testing of thin and long bone screws is essential due to screw failures during insertion and removal of the screws. Furthermore, insertion torque is an important factor in predicting fixation strength, screw pull-out force and effects of surface treatment of screws. The capability of the custom-built tester was determined using polytetrafluoroethylene and wood disc samples and bone screws. Bovine cortical bones allowed testing to the failure limit, i.e. the torque increased in long screws to the fracture limit. For 2.7 and 3.5 mm thick self-tapping cortical bone screws, the failure torques were 30-50 per cent higher than the minimum values required by the standard (1.0 and 2.3 N m respectively). The equipment provided reproducible results and fulfilled the ASTM standard very well. Preliminary testing with amorphous diamond coated bone screws showed good durability of the coating and on average 10-15 per cent lower torque values compared with uncoated screws. The equipment can be used to measure insertion and removal torques as described in the standard. Furthermore, it also allows testing of normal screws and bolts.

The defence architecture of the superficial cells of the oral mucosa.

The oral epithelium together with the saliva and its components forms a complex structure which is the first line of defence in the oral cavity. The surface of superficial cells of the oral epithelium contains ridge-like folds, microplicae (MPL), which are typical of the surfaces of areas covered with protective mucus. The role of MPL seen on the upper surface of the oral epithelial cells is still unknown. The salivary mucus gel performs a protective diffusion membrane against harmful substances and this membrane is built up by epithelial cells covered by a highly hydrated and viscous gel, where mucins constitute the scaffold. The interaction between the MPL-structure and the mucins is shown in cornea, so that mucins are expressed on the tips of the MPL of the epithelial cells. We hypothesized that the MPL architecture of oral superficial epithelial cells provides the underlying basis for mucins's protective function as well as in ocular surface. The salivary mucous barrier is required to protect the superficial cells and the MPL-structure together with membrane anchored mucin binding protein (MBP) forms the ground to this mucous barrier. So, oral mucosal barrier complex (OMBC) contains both the MBP-mucin - complex and the MPL-structure of the superficial cells. In the future, studies of the alterations of the salivary mucins and that of the MPL-structure may yield therapeutic opportunities for burning mouth syndrome and perhaps for mucositis causing by irradiation. Focus on cell surface microplication and mucins in oral mucosal biology and oral mucosal diseases is a promising avenue for future research in several ways.

Influence of physical activity and maturation status on bone mass and geometry in early pubertal girls.

This study aimed to evaluate the influence of leisure-time physical activity on the development of bone mass and density in early pubertal girls. Scores of physical activity were obtained from 242 Finnish girls (10-12 years old within Tanner Stages I-II) using a questionnaire. Bone mass and density were assessed using different densitometric techniques. At Tanner Stage I, active girls had significantly higher bone mineral mass (BMC) and areal bone mineral density (aBMD) of the whole body and cortical volumetric BMD and thickness of the tibial shaft compared with sedentary girls (P<0.05). On the other hand, the active girls at Tanner Stage II showed significantly higher values only in BMC and aBMD at the lumbar spine (P=0.017 and P=0.007, respectively). These indicated that girls at Tanner Stage I with higher leisure-time physical activity level benefited more from physical activity in terms of their bone development than their less active counterparts. Our results provide evidence that the most beneficial time for physical exercise to exhibit its effect on bone development is in the earlier pubertal period for normal school children, but the positive effect on the lumbar spine is also demonstrated in Tanner Stage II.