Embankment Quality Phase II Final Report, TR-401, 1999

Phase II research included the following: (1) develop and evaluate alternative soil design and embankment construction specifications based on soil type, moisture, density, stability, and compaction process; (2) assess various quality control and acceptance procedures with a variety of in-situ test methods including the Dual-mass Dynamic Cone Penetrometer (DCP); and (3) develop and design rapid field soil identification methods. At the start of the research, soils were divided into cohesive and cohesionless soil types, with each category being addressed separately. Cohesionless soils were designated as having less than 36% fines content (material passing the No. 200 sieve) and cohesive soils as having greater than 36% fines content. Subsequently, soil categories were refined based not only on fines content but soil plasticity as well. Research activities included observations of fill placement, in-place moisture and density testing, and dual-mass DCP index testing on several highway embankment projects throughout Iowa. Experiments involving rubber-tired and vibratory compaction, lift thickness changes, and disk aeration were carried out for the full range of Iowa soils. By testing for soil stability the DCP was found to be a valuable field tool for quality control, whereby shortcomings from density testing (density gradients) were avoided. Furthermore, critical DCP index values were established based on soil type and compaction moisture content.

Precession electron diffraction in the transmission electron Microscope: electron crystallography and orientational mapping

Precession electron diffraction (PED) is a hollow cone non-stationary illumination technique for electron diffraction pattern collection under quasikinematicalconditions (as in X-ray Diffraction), which enables “ab-initio” solving of crystalline structures of nanocrystals. The PED technique is recently used in TEMinstruments of voltages 100 to 300 kV to turn them into true electron iffractometers, thus enabling electron crystallography. The PED technique, when combined with fast electron diffraction acquisition and pattern matching software techniques, may also be used for the high magnification ultra-fast mapping of variable crystal orientations and phases, similarly to what is achieved with the Electron Backscatter Diffraction (EBSD) technique in Scanning ElectronMicroscopes (SEM) at lower magnifications and longer acquisition times.

Spermiogenesis and spermatozoon ultrastructure of the bothriocephalidean cestode Clestobothrium crassiceps (Rudolphi, 1819), a parasite of the teleost fish Merluccius merluccius (Gadiformes: Merlucciidae)

Spermiogenesis and the ultrastructure of the spermatozoon of the bothriocephalidean cestode Clestobothrium crassiceps (Rudolphi, 1819), a parasite of the teleost fish Merluccius merluccius (Linnaeus, 1758), have been studied by means of transmission electron microscopy. Spermiogenesis involves firstly the formation of a differentiation zone. It is characterized by the presence of two centrioles associated with striated rootlets, an intercentriolar body and an electron-dense material in the apical region of this zone. Later, two flagella develop from the centrioles, growing orthogonally in relation to the median cytoplasmic process. Flagella then undergo a rotation of 90° until they become parallel to the median cytoplasmic process, followed by the proximodistal fusion of the flagella with the median cytoplasmic process. The nucleus elongates and afterwards it migrates along the spermatid body. Spermiogenesis finishes with the appearance of the apical cone surrounded by the single helical crested body at the base of the spermatid. Finally, the narrowing of the ring of arched membranes detaches the fully formed spermatozoon. The mature spermatozoon of C. crassiceps is filiform and contains two axonemes of the 9 + '1' trepaxonematan pattern, a parallel nucleus, parallel cortical microtubules, and electron-dense granules of glycogen. The anterior extremity of the gamete exhibits a short electron-dense apical cone and one crested body, which turns once around the sperm cell. The first axoneme is surrounded by a ring of thick cortical microtubules that persist until the appearance of the second axoneme. Later, these thick cortical microtubules disappear and thus, the mature spermatozoon exhibits two bundles of thin cortical microtubules. The posterior extremity of the male gamete presents only the nucleus. Results are discussed and compared particularly with the available ultrastructural data on the former 'pseudophyllideans'. Two differences can be established between spermatozoa of Bothriocephalidea and Diphyllobothriidea, the type of spermatozoon (II vs I) and the presence/absence of the ring of cortical microtubules.

Genotyping microarray for CSNB-associated genes.

PURPOSE: Congenital stationary night blindness (CSNB) is a clinically and genetically heterogeneous retinal disease. Although electroretinographic (ERG) measurements can discriminate clinical subgroups, the identification of the underlying genetic defects has been complicated for CSNB because of genetic heterogeneity, the uncertainty about the mode of inheritance, and time-consuming and costly mutation scanning and direct sequencing approaches. METHODS: To overcome these challenges and to generate a time- and cost-efficient mutation screening tool, the authors developed a CSNB genotyping microarray with arrayed primer extension (APEX) technology. To cover as many mutations as possible, a comprehensive literature search was performed, and DNA samples from a cohort of patients with CSNB were first sequenced directly in known CSNB genes. Subsequently, oligonucleotides were designed representing 126 sequence variations in RHO, CABP4, CACNA1F, CACNA2D4, GNAT1, GRM6, NYX, PDE6B, and SAG and spotted on the chip. RESULTS: Direct sequencing of genes known to be associated with CSNB in the study cohort revealed 21 mutations (12 novel and 9 previously reported). The resultant microarray containing oligonucleotides, which allow to detect 126 known and novel mutations, was 100% effective in determining the expected sequence changes in all known samples assessed. In addition, investigation of 34 patients with CSNB who were previously not genotyped revealed sequence variants in 18%, of which 15% are thought to be disease-causing mutations. CONCLUSIONS: This relatively inexpensive first-pass genetic testing device for patients with a diagnosis of CSNB will improve molecular diagnostics and genetic counseling of patients and their families and gives the opportunity to analyze whether, for example, more progressive disorders such as cone or cone-rod dystrophies underlie the same gene defects.

Axial spondylometaphyseal dysplasia: Additional reports.

Axial spondylometaphyseal dysplasia (SMD) (OMIM 602271) is an uncommon skeletal dysplasia characterized by metaphyseal changes of truncal-juxtatruncal bones, including the proximal femora, and retinal abnormalities. The disorder has not attracted much attention since initially reported; however, it has been included in the nosology of genetic skeletal disorders [Warman et al. (2011); Am J Med Genet Part A 155A:943-968] in part because of a recent publication of two additional cases [Isidor et al. (2010); Am J Med Genet Part A 152A:1550-1554]. We report here on the clinical and radiological manifestations in seven affected individuals from five families (three sporadic cases and two familial cases). Based on our observations and Isidor's report, the clinical and radiological hallmarks of axial SMD can be defined: The main clinical findings are postnatal growth failure, rhizomelic short stature in early childhood evolving into short trunk in late childhood, and thoracic hypoplasia that may cause mild to moderate respiratory problems in the neonatal period and later susceptibility to airway infection. Impaired visual acuity comes to medical attention in early life and function rapidly deteriorates. Retinal changes are diagnosed as retinitis pigmentosa or pigmentary retinal degeneration on fundoscopic examination and cone-rod dystrophy on electroretinogram. The radiological hallmarks include short ribs with flared, cupped anterior ends, mild spondylar dysplasia, lacy iliac crests, and metaphyseal irregularities essentially confined to the proximal femora. Equally affected sibling pairs of opposite gender and parental consanguinity are strongly suggestive of autosomal recessive inheritance. © 2011 Wiley-Liss, Inc.

Photoreceptor types and distributions in the retinae of insectivores.

The retinae of insectivores have been rarely studied, and their photoreceptor arrangements and expression patterns of visual pigments are largely unknown. We have determined the presence and distribution of cones in three species of shrews (common shrew Sorex araneus, greater white-toothed shrew Crocidura russula, dark forest shrew Crocidura poensis; Soricidae) and in the lesser hedgehog tenrec Echinops telfairi (Tenrecidae). Special cone types were identified and quantified in flattened whole retinae by antisera/antibodies recognizing the middle-to-long-wavelength-sensitive (M/L-)cone opsin and the short-wavelength-sensitive (S-)cone opsin, respectively. A combination of immunocytochemistry with conventional histology was used to assess rod densities and cone/rod ratios. In all four species the rods dominate at densities of about 230,000-260,000/mm2. M/L- and S-cones are present, comprising between 2% of the photoreceptors in the nocturnal Echinops telfairi and 13% in Sorex araneus that has equal diurnal and nocturnal activity phases. This suggests dichromatic color vision like in many other mammals. A striking feature in all four species are dramatically higher S-cone proportions in ventral than in dorsal retina (0.5% vs. 2.5-12% in Sorex, 5-15% vs. 30-45% in Crocidura poensis, 3-12% vs. 20-50% in Crocidura russula, 10-30% vs. 40-70% in Echinops). The functional and comparative aspects of these structural findings are discussed.

Geology and ground-water resources of Cerro Gordo County, Iowa, 1970

The purposes of this report are to delineate and describe aquifers in Cerro Gordo County, evaluate the availability and quality of water in the aquifers, supply data on ground-water utilization, and determine the rate of growth and the magnitude of the cone of drawdown in the Mason City area. It includes photos and fold-out maps

The DNA-binding domain (DBD) is essential for NR2E3 dimerization and interaction with Crx

Purpose:NR2E3 (PNR) is an orphan nuclear receptor essential for proper photoreceptor determination and differentiation. In humans, mutations in NR2E3 have been associated with the recessively inherited enhanced short wavelength sensitive (S-) cone syndrome (ESCS) and, more recently, with autosomal dominant retinitis pigmentosa (adRP). NR2E3 acts in concert with the transcription factors Crx and Nrl to repress cone-specific genes and activate rod-specific genes. NR2E3 and Crx have been shown to physically interact by their DNA-binding domain (DBD), which may also be implicated in the dimerization process of the nuclear receptor. However, neither NR2E3 homodimerization nor NR2E3/Crx complex formation has been investigated in detail. Methods:In this present work, we analyzed the dimerization of the NR2E3 protein and its interaction with Crx by bioluminescence resonance energy transfer (BRET2) which utilizes Renilla luciferase (hRluc) protein and its substrate DeepBlueC as an energy donor and a mutant green fluorescent protein (GFP2) as the acceptor. We investigated, on whole intact cells, the role of NR2E3 DBD-mutations in dimerization and association with Crx. Results:We clearly showed that NR2E3 formed homodimers in HEK-293T cells. Moreover, all causative NR2E3 mutations present in the DBD of the protein showed an alteration in dimerization, except for the R76Q and the R104W mutants. Interestingly, the adRP-linked G56R mutant was the only DBD-NR2E3 mutant that showed a correct interaction with Crx. Finally, we observed a decrease in rhodospin gene transactivation for all DBD-NR2E3 mutants tested and no potentiation for the adRP-linked G56R mutant. In addition, the p.G56R mutant enhanced the transrepression of M-opsin promoter, while all other DBD-NR2E3 mutants did not repress M-opsin transactivation. Conclusions:A defect, either in the dimer formation or in the interaction of NR2E3 with Crx, leads to abnormal transcriptional activity on rhodopsin and M-opsin promoter and to an atypical retinal development; while the titration of Crx by p.G56R-NR2E3 leads to low levels of rhodopsin and M-opsin expression and may be responsible for the strong adRP phenotype.

Embankment Quality Phase IV: Application to Unsuitable Soils, TR-492, 2007

The Quality Management Earthwork (QM-E) special provision was implemented on a pilot project to evaluate quality control (QC) and quality assurance (QA) testing in predominately unsuitable soils. Control limits implemented on this pilot project included the following: 95% relative compaction, moisture content not exceeding +/- 2% of optimum moisture content, soil strength not exceeding a dynamic cone penetrometer (DCP) index of 70 mm/blow, vertical uniformity not exceeding a variation in DCP index of 40 mm/blow, and lift thickness not exceeding depth determined through construction of control strips. Four-point moving averages were used to allow for some variability in the measured parameter values. Management of the QC/QA data proved to be one of the most challenging aspects of the pilot project. Implementing use of the G-RAD data collection system has considerable potential to reduce the time required to develop and maintain QC/QA records for projects using the QM-E special provision. In many cases, results of a single Proctor test were used to establish control limits that were used for several months without retesting. While the data collected for the pilot project indicated that the DCP index control limits could be set more tightly, there is not enough evidence to support making a change. In situ borings, sampling, and testing in natural unsuitable cut material and compacted fill material revealed that the compacted fill had similar strength characteristics to that of the natural cut material after less than three months from the start of construction.

Case Study of Seasonal Variation in the Subgrade and Subbase Layers of Highway US 20, TR-516, 2008

Seasonal variations in ground temperature and moisture content influence the load carrying capacity of pavement subgrade layers. To improve pavement performance, pavement design guidelines require knowledge of environmental factors and subgrade stiffness relationships. As part of this study, in-ground instrumentation was installed in the pavement foundation layers of a newly constructed section along US Highway 20 near Fort Dodge, Iowa, to monitor the seasonal variations in temperature, frost depth, groundwater levels, and moisture regime. Dynamic cone penetrometer (DCP), nuclear gauge, and Clegg hammer tests were performed at 64 test points in a 6-ft x 6-ft grid pattern to characterize the subgrade stiffness properties (i.e., resilient modulus) prior to paving. The purpose of this paper is to present the field instrumentation results and the observed changes in soil properties due to seasonal environmental effects.

Mutation in the human homeobox gene NKX5-3 causes an oculo-auricular syndrome.

Several dysmorphic syndromes affect the development of both the eye and the ear, but only a few are restricted to the eye and the external ear. We describe a developmental defect affecting the eye and the external ear in three members of a consanguineous family. This syndrome is characterized by ophthalmic anomalies (microcornea, microphthalmia, anterior-segment dysgenesis, cataract, coloboma of various parts of the eye, abnormalities of the retinal pigment epithelium, and rod-cone dystrophy) and a particular cleft ear lobule. Linkage analysis and mutation screening revealed in the first exon of the NKX5-3 gene a homozygous 26 nucleotide deletion, generating a truncating protein that lacked the complete homeodomain. Morpholino knockdown expression of the zebrafish nkx5-3 induced microphthalmia and disorganization of the developing retina, thus confirming that this gene represents an additional member implicated in axial patterning of the retina.

Rehabilitation of Concrete Pavements Utilizing Rubblization and Crack and Seat Methods (Phase II): Performance Evaluation of Rubblized Pavements in Iowa, TR-550, 2008

This Phase II follow-up study of IHRB Project TR-473 focused on the performance evaluation of rubblized pavements in Iowa. The primary objective of this study was to evaluate the structural condition of existing rubblized concrete pavements across Iowa through Falling Weight Deflectometer (FWD) tests, Dynamic Cone Penetrometer (DCP) tests, visual pavement distress surveys, etc. Through backcalculation of FWD deflection data using the Iowa State University's advanced layer moduli backcalculation program, the rubblized layer moduli were determined for various projects and compared with each other for correlating with the long-term pavement performance. The AASHTO structural layer coefficient for rubblized layer was also calculated using the rubblized layer moduli. To validate the mechanistic-empirical (M-E) hot mix asphalt (HMA) overlay thickness design procedure developed during the Phase I study, the actual HMA overlay thicknesses from the rubblization projects were compared with the predicted thicknesses obtained from the design software. The results of this study show that rubblization is a valid option to use in Iowa in the rehabilitation of portland cement concrete pavements provided the foundation is strong enough to support construction operations during the rubblization process. The M-E structural design methodology developed during Phase I can estimate the HMA overlay thickness reasonably well to achieve long-lasting performance of HMA pavements. The rehabilitation strategy is recommended for continued use in Iowa under those conditions conducive for rubblization.

Remaining rod activity mediates visual behavior in adult Rpe65-/- mice.

Purpose: C57/Bl6, Cpfl1-/- (Cone photoreceptors function loss 1; pure rod function), Gnat1alpha-/- (rod alpha-transducin; pure cone function) and Rpe65-/-;Rho-/- double knock-out mice were studied in order to distinguish the respective contributions of the different photoreceptor (PR) systems that enable light perception and mediate a visual reflex in adult Rpe65-/- mice using a simple behavioural procedure. Methods: Visual function was estimated using a rotating automatized optomotor drum covered with vertical black and white stripes at spatial frequencies of 0.025 to 0.5 cycles per degree (cpd) in both photopic and scotopic conditions. To evaluate the contribution as well as the light intensity threshold of each PR system, we tested the mouse strains with different luminances. Results: Stripe rotation elicits head movements in wild-type (WT) animals in photopic and scotopic conditions depending on the spatial frequency, whereas Cpfl1-/- mice show a reduced activity in the photopic condition and Gnat1alpha-/- mice an almost absent response in the scotopic condition. Interestingly, a robust visual response is obtained with Rpe65-/- knockout mice at 0.075 cpd and 0.1 cpd in the photopic condition. The residual rod function in the Rpe65-/- animals was demonstrated by testing Rpe65-/-;Rho-/- mice that present no response in photopic conditions. Conclusions: The optomotor test is a simple method to estimate the visual function, and to evaluate the respective contributions of rod and cone systems. Using this test, we demonstrate that in Rpe65-/- mice, devoid of functional cones and of detectable 11-cis-retinal protein, rods mimic in part the cone function by mediating vision in photopic conditions.

Embankment Quality: Phase III, TR-401, 2002

This report describes test results from a full-scale embankment pilot study conducted in Iowa. The intent of the pilot project was to field test and refine the proposed soil classification system and construction specifications developed in Phase II of this research and to evaluate the feasibility of implementing a contractor quality control (QC) and Iowa DOT quality assurance (QA) program for earthwork grading in the future. One of the primary questions for Phase III is “Was embankment quality improved?” The project involved a “quality conscious” contractor, well-qualified and experienced Iowa Department of Transportation field personnel, a good QC consultant technician, and some of our best soils in the state. If the answer to the above question is “yes” for this project, it would unquestionably be “yes” for other projects as well. The answer is yes, the quality was improved, even for this project, as evidenced by dynamic cone penetrometer test data and the amount of disking required to reduce the moisture content to within acceptable control limits (approximately 29% of soils by volume required disking). Perhaps as important is that we know what quality we have. Increased QC/QA field testing, however, increases construction costs, as expected. The quality management-earthwork program resulted in an additional $0.03 per cubic meter, or 1.6%, of the total construction costs. Disking added about $0.04 per cubic meter, or 1.7%, to the total project costs. In our opinion this is a nominal cost increase to improve quality. It is envisioned that future contractor innovations have the potential for negating this increase. The Phase III results show that the new soil classification system and the proposed field test methods worked well during the Iowa Department of Transportation soils design phase and during the construction phase. Recommendations are provided for future implementation of the results of this study by city, county, and state agencies.

Highway Applications for Rammed Aggregate Piers in Iowa Soils, TR-443, 2003

This report describes a study to evaluate Geopier® soil reinforcement technology in transportation construction. Three projects requiring settlement control were chosen for evaluation—an embankment foundation, a box culvert, and a bridge approach fill. For each project, construction observations, in situ soil testing, laboratory material characterization, and performance monitoring were carried out. For the embankment foundation project, Geopier elements were installed within and around an abutment footprint for the new I-35 overpass at the US Highway 5/Interstate 35 interchange in Des Moines, Iowa. Although the main focus of this investigation was to evaluate embankment foundation reinforcement using Geopier elements, a stone column reinforced soil provided an opportunity to compare systems. In situ testing included cone penetration tests (CPTs), pressuremeter tests (PMTs), Ko stepped blade tests, and borehole shear tests (BSTs), as well as laboratory material testing. Comparative stiffness and densities of Geopier elements and stone columns were evaluated based on full-scale modulus load tests and standard penetration tests. Vibrating wire settlement cells and total stress cells were installed to monitor settlement and stress concentration on the reinforcing elements and matrix soil. Settlement plates were also monitored by conventional optical survey methods. Results show that the Geopier system and the stone columns performed their intended functions. The second project involved settlement monitoring of a 4.2 m wide x 3.6 m high x 50 m long box culvert constructed beneath a bridge on Iowa Highway 191 south of Neola, Iowa. Geopier elements were installed to reduce total and differential settlement while ensuring the stability of the existing bridge pier foundations. Benefits of the box culvert and embankment fill included (1) ease of future roadway expansion and (2) continual service of the roadway throughout construction. Site investigations consisted of in situ testing including CPTs, PMTs, BSTs, and dilatometer tests. Consolidated drained triaxial compression tests, unconsolidated undrained triaxial compression test, oedometer tests, and Atterberg limit tests were conducted to define strength and consolidation parameters and soil index properties for classification. Vibrating wire settlement cells, total stress cells, and piezometers were installed for continuous monitoring during and after box culvert construction and fill placement. This project was successful at controlling settlement of the box culvert and preventing downdrag of the bridge foundations, but could have been enhanced by reducing the length of Geopier elements at the ends of the box culvert. This would have increased localized settlement while reducing overall differential settlement. The third project involved settlement monitoring of bridge approach fill sections reinforced with Geopier elements. Thirty Geopier elements, spaced 1.8 m apart in six rows of varying length, were installed on both sides of a new bridge on US Highway 18/218 near Charles City, Iowa. Based on the results of this project, it was determined that future applications of Geopier soil reinforcement should consider extending the elements deeper into the embankment foundation fill, not just the fill itself.