Unaffected Sites Research Articles

Prolonged shear stress and KLF2 suppress constitutive proinflammatory transcription through inhibition of ATF2

AbstractAbsence of shear stress due to disturbed blood flow at arterial bifurcations and curvatures leads to endothelial dysfunction and proinflammatory gene expression, ultimately resulting in atherogenesis. KLF2 has recently been implicated as a transcription factor involved in mediating the anti-inflammatory effects of flow. We investigated the effect of shear on basal and TNF-α–induced genomewide expression profiles of human umbilical vein endothelial cells (HUVECs). Cluster analysis confirmed that shear stress induces expression of protective genes including KLF2, eNOS, and thrombomodulin, whereas basal expression of TNF-α–responsive genes was moderately decreased. Promoter analysis of these genes showed enrichment of binding sites for ATF transcription factors, whereas TNF-α–induced gene expression was mostly NF-κB dependent. Furthermore, human endothelial cells overlying atherosclerotic plaques had increased amounts of phosphorylated nuclear ATF2 compared with endothelium at unaffected sites. In HUVECs, a dramatic reduction of nuclear binding activity of ATF2 was observed under shear and appeared to be KLF2 dependent. Reduction of ATF2 with siRNA potently suppressed basal proinflammatory gene expression under no-flow conditions. In conclusion, we demonstrate that shear stress and KLF2 inhibit nuclear activity of ATF2, providing a potential mechanism by which endothelial cells exposed to laminar flow are protected from basal proinflammatory, atherogenic gene expression.

Diffusion-weighted MR imaging of viral encephalitis

The aim of this study was to evaluate the role of diffusion-weighted imaging (DWI) in the diagnosis of viral encephalitis and its relationship with the stage of the illness. We performed conventional magnetic resonance imaging (MRI) including T1-W, T2-W and fluid attenuated inversion recovery (FLAIR) sequences and DWI in 18 patients with viral encephalitis diagnosed on the basis of laboratory, clinical and radiologic findings. Based on the qualitative and quantitative comparison of the conventional MRI and DWI, the patients were divided into three groups. Apparent diffusion coefficient (ADC) values of the involved and contralateral normal brain tissues were computed and compared for each group. The degree of correlation between the time (TI) from the onset of neurologic symptoms to the MR examination and ADC values was determined. In group I (n=11) DWI was superior to conventional MRI in detecting the encephalitic involved sites and in depicting the borders of the encephalitic lesions. In group II (n=4) DWI was similar to conventional MRI. In group III (n=3) conventional MRI was superior to DWI. Mean ADC values of affected versus contralateral normal brain tissues were 0.458+/-0.161 x 10(-3) versus 0.86+/-0.08 x 10(-3) in group I, 0.670+/-0.142 x 10(-3) versus 0.93+/-0.07 x 10(-3) in group II, and 1.413+/-0.211 x 10(-3) versus 1.05+/-0.06 x 10(-3) in group III. Patients in group I had significantly lower ADC values than those in group II, while patients in group III had the highest ADC values (P<0.05). The ADC values were significantly lower in the affected sites than in the unaffected sites of patients in groups I and II, but were significantly higher in the affected sites than in the unaffected sites of patients in group III (P<0.05). There was an excellent correlation between ADC values and duration of the disease (r=0.874, P=0.01). DWI is superior to other conventional diagnostic MR sequences in the detection of early viral encephalitic lesions and depiction of the lesion borders and, in combination with other sequences, DWI may contribute to the determination of the disease phase.

Hydrocarbon contamination of a typical mangrove floor in Niger Delta, Nigeria

The impact of petroleum hydrocarbons on the floor of a mangrove system located on one of the intertidal lands behind the barrier islands of Niger Delta basin of southern Nigeria was evaluated following the Obiafu-14 oil spillage. The area covered by the oil spill was delimited by reconnaissance and soils were sampled by grid method from the bottom and middle slopes of oil-affected and unaffected (control) sites. Soil samples were later brought to the laboratory where total hydrocarbon content was determined by extraction and spectrophotometric techniques. A lower pH range of 4.28–4.36 in the oil impacted soils meant that the site was more acidic; a higher moisture content of 33.34% and lower electrical conductivity (EC) of 31.75μS/cm in the affected area presupposed oxygen deprivation and lower salt content respectively. On the strength of the baseline data on the study area and evidence from the uncontaminated, geographically similar control site, it can be said that the Obiafu-14 oil spillage had severely contaminated the affected mangrove floor. This is corroborated by a total extractable hydrocarbon content of 6.32×103 ± 344 mg/kg (no overlap in Standard Error at 95% Confidence Limit), and the abysmal degradation of the mangrove system at the site. Although soils of the mangrove floor under consideration may well be of limited cultivation value on account of the people’s dual occupation in fishing and farming, the presence of such levels of petroleum hydrocarbons at the study site might hamper agricultural productivity.

Texture Analysis of Post Breast Cancer Lymphedema Ultrasound Images Obtained Using a Portable Device—A Pilot Study

Lymphedema is a common complaint of post-breast cancer treatment. Some prior ultrasound imaging studies of lymphedema-affected upper extremities focused upon estimating the thickness of the dermis and subcutis areas. In contrast, however, the reliability and validity of texture features derived from ultrasound images obtained using a low-cost portable ultrasound device have not been reported. This study examined the reliability and validity of the first and second order quantitative image texture measures, average pixel intensity and entropy, and compared their site-specific correlations. A total of 10 ultrasound images of an ipsilateral affected elbow and the corresponding unaffected contralateral elbow were obtained using a portable ultrasound device. A Graphical User Interface software package for image analysis was developed and tested. Entropy and average pixel intensity at six adjacent regions of interest corresponding to the dermis and subcutis areas were calculated. Entropy had smaller coefficients of variation for each image compared to average pixel intensity with the intracluster correlation reliability coefficient for entropy being twice that of average pixel intensity. Both measures discriminated between the affected and unaffected sites (p < 0.001). However, the correlation between the two measures differed for affected (r = -0.060, p = 0.667) and unaffected (r = +0.595, p < 0.001) sites. The second order measure entropy is preferable to that of the first order average pixel intensity as a measure of image texture to characterize ultrasound images of extremity lymphedema.

Gene Expression Profiling in Seminoma and Nonseminoma

Gene expression profiles of seminoma were compared with nonseminoma to get insights into tumorigenesis. Eleven testicular tumor biopsies (five pure seminoma, six nonseminoma; pT1N0M0 to pT2N2M1) and biopsies from unaffected sites were analyzed once per patient using a macroarray (1,176 genes). On the same patients, six genes were validated using real-time quantitative (RTQ) polymerase chain reaction (PCR). Additionally, in a separate cohort of 19 patients, 24 genes selected from the macroarray were measured using RTQ-PCR. (1) The agreement in gene expression was 94% between the two methods and two different patient cohorts. (2) Two features in gene expression were independent of the tumor entity: Most changes of gene expression occurred in five functional groups like "cell cycle" and "apoptosis." Genes within these groups were almost similarly (> 80%) up- or downregulated. (3) Nonseminoma were characterized by downregulated genes (75%), but in seminoma, upregulated genes (64%) prevailed. Furthermore, 64.4% of those genes that were differentially expressed in both tumor entities were usually upregulated in seminoma but downregulated in nonseminoma. A reverse pattern was found in 24.4% of such genes. Eleven percent of these genes showed a similar up- or downregulation in gene expression in both tumor entities. Seminoma in this preliminary study can be differentiated from nonseminoma due to almost opposing gene expression profiles (89% of the significantly differentially expressed genes) and are in line with the histological discrimination of both tumor entities. Underlying mechanisms and implications regarding the origin and tumor progression of both entities are discussed.

Mobility and Persistence of Petroleum Hydrocarbons in Peat Soils of Southeastern Mexico

Spilled crude petroleum from oil wells contains numerous hydrocarbons, some of which are toxic and threaten life. We have studied the mobility and persistence of hydrocarbons in waterlogged soils that contain large proportions of fermented organic matter (Histosols) and large concentrations of dissolved organic carbon (DOC) in the State of Tabasco, Mexico. We sampled soil and phreatic water at sites polluted by oil spills for several decades, as well as at sites that had only recently (few weeks) been polluted, and compared their hydrocarbon contents with those of unaffected sites in the same area. Samples were analyzed for 16 non-alkylated polyaromatic hydrocarbons (PAHs) and n-alkanes from nC9 to nC34. The spilled hydrocarbons had remained predominantly in the organic surface horizons of the soil where spillage occurred; there was little evidence of movement within the soil. The fraction of low molecular weight compounds was larger at sites of recent spills than where spills happened several decades ago. Nevertheless, sites of old spills still contained large concentrations of hydrocarbons, among which those of low molecular weight represented from 30 to 49% of total PAHs and from 50 to 84% of total n-alkanes, indicating that volatilization or microbial degradation is slow in these soils. In the peat horizons the measured organic carbon partition coefficients (K oc ) for the higher molecular weight PAHs were consistently smaller than those estimated by empirical equations by up to two orders of magnitude. The dissolved organic carbon of these peat soils seems to influence this behavior. At sites of old spills, partition coefficients for the PAHs were larger than at sites of recent spills.

Familial Segmental Neurofibromatosis

Segmental neurofibromatosis is considered to be the result of postzygotic NF1 gene mutations. We present a family in which the proband has generalized neurofibromatosis 1, whereas members of previous generations manifest segmental skin lesions. All, including the clinically asymptomatic grandmother, carry the same haplotype. This is the only case in the literature in which a parent with segmental skin findings has a child with full-blown neurofibromatosis 1 disease. The genetic mechanisms underlying this association are discussed. This family can be further investigated by examination of tissue samples from affected and unaffected sites for mutations.

Influence of the solid-state modification of MCM-22 with Cs on toluene disproportionation

The catalytic performance of MCM-22 modified with Cs to different extents by Solid-State Ion Exchange has been investigated in toluene disproportionation. It was suggested that protons on the external surface were preferentially replaced by Cs cations. It was observed that the Cs modifications did not provide improved p-xylene selectivity at conversions close to that of the H-form. The remaining unaffected proton sites in the 10 MR channels are considered as responsible for secondary xylene isomerization proceeding upon toluene disproportionation.

Distribution of anthropogenic mercury in French Guyana river sediments downstream from gold mining sites

Total mercury concentrations in French Guyana river sediments were determined near gold mining sites and compared to unaffected reference sites. In small meter-size rivers the anthropogenic impact is localized and characterised by high total mercury peak concentrations in the mg/kg dry weight range (St. Elie site). The mercury load is more diluted in larger water streams such as the Inini river system where a mercury gradient was observed along a 50 km flow path downstream with 510, 295, 261 ng/g total mercury concentrations in the sediments at 0, 35 and 50 km distance from the Dorlin mining site. Sediments situated upstream the gold mining sites compared to the geochemical background concentrations indicating that the impact related to atmospheric deposition is relatively insignificant. The mercury content of upstream sediments compare to the mercury load of unaffected soils and river sediments collected in distant virgin sites (less than 400 ng/g total mercury).

Impacts of the Jessica oil spill on intertidal and shallow subtidal plants and animals

Densities of fishes, invertebrates and plants at rocky intertidal and shallow subtidal sites were censused 1–2 days prior to the Jessica oil spill and compared with information obtained for the same sites one month after the spill, both for sites impacted by oil and unaffected reference sites. While the availability of pre-spill data made this analysis one of the most powerful to date for testing impacts of oil on shoreline environments, no clear changes attributable to oiling could be identified. Discharged oil appeared to cause very little impact in the intertidal and shallow subtidal zones, with such impacts lying within the range of natural spatial and temporal variation at sites investigated. Factors considered to minimize impact in Galápagos included paucity of fully sheltered shores in spill path, moderate wave action, warm temperature, high levels of sunlight, and mixing of bunker oil with diesel.

Effect of anoxia and high sulphide concentrations on heterotrophic microbial communities in reduced surface sediments (Black Spots) in sandy intertidal flats of the German Wadden Sea

Black reduced sediment surfaces (Black Spots) in sandy intertidal flats of the German Wadden Sea (southern North Sea) are characterised by elevated sulphide concentrations (up to 20 mM) and low redox potentials. It is assumed that the appearance of Black Spots is linked to elevated levels of organic matter content within the sediments. In order to establish the effect of high substrate and sulphide concentrations on the heterotrophic microbial communities in Black Spot sediments, bacterial abundances and the potential C-source utilisation patterns of microbial communities were compared in natural and artificially induced Black Spots and unaffected control sites. Bacterial numbers were estimated by direct counts and the most probable number technique for different physiological groups, while patterns of C-substrate utilisation of entire aerobic microbial communities were assessed using the Biolog™ sole-carbon-source-catabolism assay. Bacterial abundances at Black Spot sites were increased, with increases in mean cell numbers, more disperse data distributions and more extreme values. Substrate utilisation patterns of aerobic microbial communities were significantly different in Black Spot sediment slurries, showing diminished richness (number of C-sources catabolised) and substrate diversity (Shannon diversity index) in comparison to unaffected sites. Principal component analysis clearly discriminated Black Spot utilisation patterns from controls and indicated that microbial communities in individual Black Spot sites are functionally diverse and differ from communities in oxidised surface sediments and reduced subsurface sediments at control sites. This work suggests that potentially negative effects on microbial communities in Black Spot sediments, through anoxia and high sulphide concentrations, are balanced by the stimulating influence of substrate availability, leading to comparable or higher bacterial numbers, but lower functional microbial diversity of aerobic microbial communities.

The role of gustatory flushing in Frey's syndrome and its treatment with botulinum toxin type A.

After parotid surgery, gustatory sweating and flushing occur more frequently, the former reportedly in 15-100% of cases, while no reliable data are available for the latter. Although botulinum toxin (BoNT) is effective in controlling sweating, little is known about its effect on flushing. In 17 patients suffering from Frey's syndrome after parotid surgery, we studied the gustatory flushing phenomenon as compared to gustatory sweating, analyzing their frequency, area, type of stimulus and response to BoNT administration. Cutaneous blood flow (CBF) was monitored by laser Doppler flowmetry (LDF) on affected and unaffected areas of the cheek in basal conditions and after meals, before and then 1 month after starting the BoNT injections. The Minor test was used to identify the sweating area. Flushing was observed in 7 of 17 patients after masticatory activity, spicy meals or citrus fruits. No clinical data correlated with any presence of flushing. Flushing regressed completely after BoNT administration and CBF reached similar values in the affected and unaffected sites. No adverse effects were observed. BoNT administration proved an effective and safe treatment for gustatory sweating and flushing in patients with Frey's syndrome.

Infectious agents associated with respiratory disease in pheasants

In a case-control study of the infectious agents associated with natural outbreaks of respiratory disease in pheasants, 28 batches of birds from sites affected by disease and eight batches of...

Mensuration of cervical vertebral ratios in Doberman pinschers and Great Danes.

Canine cervical vertebral instability is a complex syndrome involving cervical spinal cord compressive lesions. Doberman pinschers and Great Danes are the most commonly affected breeds. Measurements of vertebral canal diameter (VCD), vertebral body height (H), and vertebral body length (L) were made from C3-C7 in 24 Doberman pinschers and 8 Great Danes by using digital and analog techniques. Significant differences between affected and unaffected sites were noted for Doberman pinschers at C6 (P = 0.039) and C7 (P = 0.027) using analog measurements of VCD/H, and Great Danes at C6 using analog (P = 0.041) and digital (P = 0.004) measurements of VCD/L. Cervical vertebral ratios have potential as a breed-specific screening tool for cervical vertebral instability and warrant longitudinal studies.

The Predilection Sites for Chronic Atopic Dermatitis Do Not Show Any Special Functional Uniqueness of the Stratum corneum

The functional deficiencies of the stratum corneum (SC) have been well documented in the skin of atopic dermatitis (AD) patients. AD tends to affect certain predilection areas chronically such as the face, neck and the flexural folds of the extremities. From the paucity of the information about the functional characteristics of the SC in such skin areas, we conducted biophysical measurements at the predilection sites of AD, i.e. the cheek, anterior surface of the neck, nape, antecubital fossa and popliteal fossa and relatively uninvolved sites such as the elbow, mid volar forearm, knee and mid calf in 34 AD patients and in 40 normal individuals. Reflecting the great interlocation variations in the SC noted even in the normal individuals, we found location-dependent as well as severity-dependent changes of SC function in the AD patients. Thus, the values of transepidermal water loss (TEWL) obtained on the cheek and, unexpectedly, the elbow were quite high, whereas those on the poplieal fossa were very low; even those values recorded on the lesional skin of the latter of the AD patients were significantly lower than those of the cheek of normal healthy individuals (p < 0.05). Moreover, the TEWL values measured at the nonlesional skin of AD patients were much closer to those of the normal healthy skin than to those of the lesional skin in most areas. Similarly, the SC hydration state that is influenced by the amount of skin surface lipids showed prominent location-dependent changes in addition to a severity-dependent change. It was much higher even in the lesional skin of the face and neck than in the normal skin of the extensor surface of the limbs. The corneocyte size, which inversely correlates with the turnover rate of the SC, was larger even on the lesional skin of the extremities than that of the normal cheek. It is only in skin surface pH that such location dependency was not demonstrable; it uniformly revealed higher values in the lesional and nonlesional skin of AD patients regardless of the sites. From these comparative data obtained at the predilection sites and relatively unaffected sites of AD, we think that those functional abnormalities of the SC reported in AD patients simply reflect the severity of underlying inflammatory changes rather than inherently defective SC functions.

Porites corals as recorders of mining and environmental impacts: Misima Island, Papua New Guinea

In 1989 open-cut gold mining commenced on Misima Island in Papua New Guinea (PNG). Open-cut mining by its nature causes a significant increase in sedimentation via the exposure of soils to the erosive forces of rain and runoff. This increased sedimentation affected the nearby fringing coral reef to varying degrees, ranging from coral mortality (smothering) to relatively minor short-term impacts. The sediment associated with the mining operation consists of weathered quartz feldspar, greenstone, and schist. These rocks have distinct chemical characteristics (rare earth element patterns and high abundances of manganese, zinc, and lead) and are entering the near-shore environment in considerably higher than normal concentrations. Using laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS), we analyzed eight colonies (two from high sedimentation, two transitional, two minor, and two unaffected control sites) for Y, La, Ce, Mn, Zn, and Pb. All sites show low steady background levels prior to the commencement of mining in 1988. Subsequently, all sites apart from the control show dramatic increases of Y, La, and Ce associated with the increased sedimentation as well as rapid decreases following the cessation of mining. The elements Zn and Pb exhibit a different behavior, increasing in concentration after 1989 when ore processing began and one year after initial mining operations. Elevated levels of Zn and Pb in corals has continued well after the cessation of mining, indicating ongoing transport into the reef of these metals via sulfate-rich waters. Rare earth element (REE) abundance patterns measured in two corals show significant differences compared to Coral Sea seawater. The corals display enrichments in the light and middle REEs while the heavy REEs are depleted relative to the seawater pattern. This suggests that the nearshore seawater REE pattern is dominated by island sedimentation. Trace element abundances of Misima Island corals clearly record the dramatic changes in the environmental conditions at this site and provide a basis for identifying anthropogenic influences on corals reefs.

International approach to assessing soil quality by ecologically-related biological parameters

Soil quality represents an integral value of the compositional structures and natural functions of soil in relation to soil use and environmental conditions on site. Among the indigenous soil components, different organisms and especially microorganisms play a key role in ecologically important biogeochemical processes. In that way, soil microorganisms contribute to the maintenance of the matter and energy transfer in terrestrial environments. Under stress conditions caused by adverse anthropogenic effects such as dissemination of chemical pollutants, the development and biochemical activities of soil microorganisms undergo several alterations. To prevent negative ecological consequences, microbiologically-related parameters should be involved in the indication of soil quality. In order to indicate some reliable indicators, an international project was established in which scientists from the Czech Republic, Hungary, Russia, Slovak Republic and Germany were participating. Standardized methods for selected microbiological and biochemical soil parameters were repeatedly applied in the individual laboratories on soil samples from rather unaffected and anthropogenically-affected soil sites (49 in total). After evaluation of more than 20 individual parameters it was concluded that N2-fixing bacteria, total microbial biomass, soil respiration, dehydrogenase activity, and perhaps also the humification activity of soil microorganisms could serve as sensitive indicators of soil quality. Further research is necessary, however, to recognize critical limits with respect to the different soil types and uses. Seasonal oscillations in ecologically-related soil biological activities which regularly occur should be appropriately respected.

Spatial distribution and histogenesis of colorectal Paneth cell metaplasia in idiopathic inflammatory bowel disease.

Colorectal Paneth cell metaplasia (PCM) is known to be a sign of idiopathic inflammatory bowel disease (IBD), although its distribution and histogenesis are not fully understood. Objectives of this research were to investigate the spatial distribution of PCM in IBD and other forms of colitis (non-IBD), and to find stimuli causing PCM. We studied multiple biopsy specimens from 181 patients with ulcerative colitis (UC), 159 with Crohn's disease (CD), 448 with non-IBD, and 78 normal controls. Paneth cell metaplasia frequency, at each colorectal site, was evaluated to find possible differences among diseases, phases of activity, and extents of disease. In non-IBD and controls, PCM was rarely (0-1.9%) seen at distal sites, but frequently (up to 48.7%) found at the ascending colon and cecum (P < 0.001). Paneth cell metaplasia frequency was significantly higher in IBD than in non-IBD patients and controls at distal sites (P < 0.001), but did not differ significantly between UC and CD, or among active, resolving, and quiescent phases. In UC, proctitis and left-sided colitis rarely displayed PCM at unaffected sites. Multiple logistic regression analysis revealed that PCM was positively associated with crypt distortion and mononuclear cell infiltration (P < 0.005), but negatively or not significantly associated with crypt atrophy, mucin depletion, acute inflammation, or phase of activity. Paneth cell metaplasia is a non-specific phenomenon in the proximal colon, but distal PCM, which occurs exclusively in affected mucosa, is a useful marker indicating IBD, even in the inactive phase. Regression analysis suggests that repair and regeneration may be the most potent stimuli causing PCM.

Absence of measles virus and canine distemper virus transcripts in long-term bone marrow cultures from patients with Paget’s disease of bone

The presence of intranuclear inclusions in pagetic osteoclasts with similar characteristics to those seen in some slow viral diseases has lead to the hypothesis that Paget’s disease is caused by a similar infection. Bone marrow aspirates from seven patients with hemipelvic Paget’s disease were taken from both sides of the pelvis and cultured under identical conditions. RNA was extracted after approximately 2 weeks of culture and reverse transcriptase-linked polymerase chain reaction used to investigate the expression of measles and canine distemper virus RNA. We were unable to detect the presence of measles virus (MV) or canine distemper virus (CDV) transcripts in bone marrow cultures from either affected or unaffected sites in any of our patients. The results of this study do not support a role for MV or CDV in the pathogenesis of Paget’s disease of bone.

Relative deficiency in IgA production by duodenal explants from German shepherd dogs with small intestinal disease

Matched samples of serum, saliva and tears were collected from four groups of dogs; two of the groups were German shepherd dogs (GSDs) either with (Group 1) or without (Group 4) a variety of small intestinal disorders; the remaining two groups were dogs of other breeds, again with (Group 2) or without (Group 3) small intestinal disease. Capture ELISAs were used to measure IgG, IgM, IgA and albumin concentrations within these samples; intestinal humoral immune status of clinical cases was assessed by quantifying immunoglobulin production from duodenal explant cultures. There were no significant differences in IgG, IgM or IgA concentrations in serum, saliva or tears between the different groups of dog. Moreover, no significant differences were noted between groups for IgG, IgM and IgA salivary and tear secretory indices. IgA production by 24-h explant cultures was significantly lower in GSDs compared with non-GSDs with small intestinal disease (groups 1 and 2, respectively), but the numbers of lamina propria IgA + plasma cells in duodenal biopsies were not different between groups. These results suggest that there may be a relative deficiency in local IgA secretion in GSDs with small intestinal enteropathies, which is not reflected in either serum IgA concentrations, or in secretion at unaffected mucosal sites. It remains to be determined whether such a deficiency is a breed-related primary defect, or whether it arises secondary to the pathological processes within the intestinal mucosa.