Mechanical properties of the human lumbar anterior longitudinal ligament.

A new technique incorporating a motion analysis system and a materials testing machine was used to investigate regional differences in the tensile mechanical properties of the lumbar spine anterior longitudinal ligament (ALL). Bone-ALL-bone specimens were prepared from young human cadaveric motion segments with no disc or bony pathology. Each specimen was distracted until failure at a constant crosshead displacement rate of 2.5 mm s-1 (approximately 1.0% strain per second). Strains were evaluated from digitized video recordings of markers attached to the ALL at 12 sites along its length and width, including the ligament substance and insertions. The 'overall' strain in the ligament was calculated from the outermost pairs of markers along the ligament length. The average tensile strength, the 'overall' tensile modulus and the 'overall' strain of the ALL at failure were 27.4 MPa (S.D. 5.9), 759 MPa (S.D. 336) and 4.95% (S.D. 1.51), respectively. Large and significant variations in the strains were present along the width and length of the ALL. Peak substance strains were over twofold greater than peak strains at the ligament insertion sites, whereas across the ligament width, peak strains in the outer portion of the ligament were over 40% greater than in the central region. Failure consistently occurred in the ligament mid-substance and ultimate strains at the ligament failure site averaged 12.1% (S.D. 2.3). These results indicate that the strains are highly nonuniform in the normal ALL.     

On the identification of the most suitable traits for plant functional trait analyses

Within the past few years plant functional trait analyses have been widely applied to learn more about the processes and patterns of ecosystem development in response to environmental changes. These approaches are based on the assumption that plants with similar ecologically relevant trait attributes respond to environmental changes in comparable ways. Several methods have been described on how to analyse a priori defined trait sets with respect to environment. Irrespective of the statistical methods used to contrast ecosystem responses and environmental conditions, each functional trait approach depends strongly on the initial trait set. In nearly all recent studies on functional trait analysis a test, if a trait is responsible, is applied independently from the core analysis. In the current study we present a method that extracts those traits from a wider set of traits which are optimal for describing the ecosystem response to a given environmental gradient. This was done by the use of iterative three‐table ordination techniques with each possible trait combination. We further concentrated on the effect of the inclusion of too many traits in such analyses. As examples the method was applied to three long term studies on abandoned arable fields. The approach was validated by comparing the results with literature‐knowledge on arable field succession. Although the trait pre‐selection was only based on a statistical procedure, our method was able to identify all relevant processes of ecosystem responses. All three sites show comparable ecosystem responses; the importance of the competitive ability of plants was highlighted. We further demonstrated that the use of too many traits results in an over‐fitting of the trait‐environment model. The presented method of iterative RLQ‐analyses is adequate to identify responding traits to environmental changes: the discovered processes of successional development of abandoned arable fields are consistent with our knowledge from the literature.     

PlantRGS: a web server for the identification of most suitable candidate reference genes for quantitative gene expression studies in plants

Normalization of quantitative gene expression data with a suitable reference gene is essential for accurate and reliable results. However, the availability and choice of most suitable reference gene(s) showing uniform expression across all the experimental conditions remain a drawback. We have developed a web server, PlantRGS (http://www.nipgr.res.in/PlantRGS), for the identification of most suitable candidate reference gene(s) at the whole-genome level using microarray data for quantitative gene expression studies in plants. Microarray data from more than 11 000 tissue samples for nine plant species have been included in the PlantRGS for meta-analysis. The web server provides a user-friendly graphical user interface-based analysis tool for the identification of most suitable reference genes in the selected plant species under user-defined experimental conditions. Various parameter options and output formats will help users to investigate desired number of most suitable reference genes with wide range of expression levels. Validation of results revealed that novel reference genes identified by the PlantRGS outperforms the traditionally used reference genes in terms of expression stability. We anticipate that the PlantRGS will provide a platform for the identification of most suitable reference gene(s) under given experimental conditions and facilitate quantitative gene expression studies in plants.     

Fluorescence-based AFLPs occur as the most suitable marker system for oilseed rape cultivar identification

Three different types of molecular markers, RAPD, SSR and fluorescence-based AFLP, were evaluated and compared for their ability to identify oilseed rape cultivars. The direct comparison of RAPD, SSR and AFLP approaches in cultivar identification showed that the AFLP methodology detected polymorphisms more efficiently than either RAPD or SSR methods. For the characterisation of six oilseed rape cultivars, 60 RAPD primers were tested and only eight of them (14%) detected sufficient levels of polymorphism. Five microsatellites out of fifteen tested were polymorphic, but in all loci, except one, only two different alleles were detected. This result indicated the limited degree of polymorphism found in Brassica napus. Each of the six tested AFLP combinations detected polymorphisms, the best combination (M-CAA/E-ACT) had 26% polymorphic peaks from a total of 90 peaks and could distinguish the analysed cultivars and 4 out of 5 core lines of cultivars. The results presented show that florescence-based AFLP is, for the purposes of oilseed rape cultivar fingerprinting, a more suitable approach than either RAPD or SSR.     

Rheological properties of the human lumbar spine ligaments

The purpose of this study is to provide a better understanding of the rheological properties of the lumbar spinal ligaments under subfailure physiological loads. Non-destructive tests including an hysteresis experiment, stress-relaxation and stepwise load-relaxation tests were used to investigate the time-dependent properties of the interspinous-supraspinous ligament complex. Using a reduced relaxation function, the viscoelastic behaviour over the experimental time-scale was described by a linear function of the logarithm of time. Internal damping of ligament substance dissipates about 36% of the mechanical energy applied during physiological loading. Local elastic stiffness is found to be two to four times global stiffness of the bone-ligament-bone complex. These physical parameters (stiffness, energy dissipation, hysteresis, relaxation, etc) can be used to improve computer models of the lumbar spinal column.     

Atypical localization of myenteric ganglia in the human appendical wall: a comparative study with animal appendix

The presence of well developed appendices in some animals when compared to humans has led to speculation that appendix is a vestigial organ. Increasing number of studies have revealed that the appendix serves as an important organ in humans. The function of animal appendix, and the differences between species remain poorly understood. In this study we examined human myenteric plexus and compared them with animal studies. Appendices were obtained from five young adults in which the appendix was found to be normal after removal. Fixed appendix cryosections were examined by immunofluorescence methods using neuronal marker antibodies to neurofilaments and beta III tubulin. Both antibodies stained myenteric ganglia which were arranged in an apparently irregular pattern in human appendix wall. We observed unexpected localization of myenteric ganglia in the subserosa often accompanied by rarely occurring ganglia in the longitudinal muscle layer. These ganglia were of different sizes and shapes and unequally distributed under a thin layer of serosa. Our findings raise many questions about the possible role of irregular and atypical myenteric ganglia localization in relation to altered motility and subsequent pathogenesis of the appendix in inflammatory disease in humans. On the other hand, studies of the literature have revealed simplicity in the organization of myenteric plexus, e.g., in well-developed rabbit appendix. In addition, appendicitis in animals is restricted to in apes with similarly shaped appendix to humans.     

Lectins as markers of endothelial cells: comparative study between human and animal cells

Vascular endothelial cells were labelled with 10 vegetal lectins and 3 more monoclonal antibodies antiblood group ABO substances, in major organs of 14 common laboratory animals. After fixation in PLPa and paraffin embedding, cells were examined to determine their likeness to human cells. The most interesting reactive used was EEA, whose positivity defines upper mammalians. Blood B substance positivity and CSA negativity defines primates among which man is unique and defined by UEA I positivity and variability in ABO substance. CSA positivity defines non-primate upper mammalians. Rodents and birds were negative with all reactives tested. From the histochemical point of view, the animals closest to humans are monkeys, followed by swine and oxen, then by cat and dog and lastly by sheep. Rodents appear unrelated to humans in this system.     

The comparative ultrastructure of the epididymis in monkeys and man: A search for a suitable animal model for studying epididymal physiology in primates

To assess both the need and potential of different primates for studying human epididymal function, the ultrastructure of the epididymis in several New and Old World monkeys has been compared with that of man. Sexually mature monkeys of six species were used; three talapoin monkeys, two pig-tail macaques, one patas monkey, one capuchin, one spider monkey, and four common marmosets. Samples of human epididymis were obtained from men undergoing vasectomy. Tissue was examined by light and electron microscopy and observations were quantified using image analysis. The primate epididymis displayed several ultrastructural features not observed in other mammals. These included the presence of small membrane-bound granules in the infranuclear cytoplasm of principal cells, and a close association of blood capillaries with the basal lamina and mitochondria-rich cells. Differences were apparent in the number and volume of organelles in principal cells from different regions of the epididymis and between species. Epididymal tissue in man showed a much greater ultrastructural diversity than that of monkeys. The significance of these results is discussed in relation to the need for an animal model for studying the primate epididymis.     

Geometric and mechanical properties of human cervical spine ligaments

This study characterized the geometry and mechanical properties of the cervical ligaments from C2-T1 levels. The lengths and cross-sectional areas of the anterior longitudinal ligament, posterior longitudinal ligament, joint capsules, ligamentum flavum, and interspinous ligament were determined from eight human cadavers using cryomicrotomy images. The geometry was defined based on spinal anatomy and its potential use in complex mathematical models. The biomechanical force-deflection, stiffness, energy, stress, and strain data were obtained from 25 cadavers using in situ axial tensile tests. Data were grouped into middle (C2-C5) and lower (C5-T1) cervical levels. Both the geometric length and area of cross section, and the biomechanical properties including the stiffness, stress, strain, energy, and Young's modulus, were presented for each of the five ligaments. In both groups, joint capsules and ligamentum flavum exhibited the highest cross-sectional area (p < 0.005), while the longitudinal ligaments had the highest length measurements. Although not reaching statistical significance, for all ligaments, cross-sectional areas were higher in the C5-T1 than in the C2-C5 group; and lengths were higher in the C2-C5 than in the C5-T1 group with the exception of the flavum (Table 1 in the main text). Force-deflection characteristics (plots) are provided for all ligaments in both groups. Failure strains were higher for the ligaments of the posterior (interspinous ligament, joint capsules, and ligamentum flavum) than the anterior complex (anterior and posterior longitudinal ligaments) in both groups. In contrast, the failure stress and Young's modulus were higher for the anterior and posterior longitudinal ligaments compared to the ligaments of the posterior complex in the two groups. However, similar tendencies in the structural responses (stiffness, energy) were not found in both groups. Researchers attempting to incorporate these data into stress-analysis models can choose the specific parameter(s) based on the complexity of the model used to study the biomechanical behavior of the human cervical spine.     

Inhibition of trypsins and chymotrypsins from different animal species: a comparative study

The effect of 3 purified trypsin inhibitors and 4 legume seed extracts on teh trypsins and chymotrypsins of the activated pancreata of 11 animal species, including man, was measured. The activation was performed by either homologous enterokinase or by bovine trypsin. Several trypsinogens were not activated by the latter. Rabbit trypsin was the most sensitive to all inhibitor preparations, while the human trypsin was the most resistant, except to the black bean extract. The response of the chymotrypsins was more variable and those of capybara and rabbit showed extreme sensitivity. Considerable differences between the extracts of black and white garden beans, both Phaseolus vulgaris, with respect to their reactivity toward different animal enzymes were detected. No relation between relative pancreas weight and susceptibility toward soybean trypsin inhibitor could be observed.     

Species identification and confirmation of human and animal cell lines: a PCR-based method

Misidentification and cross-contamination of cell lines are major problems of cell cultures that can make scientific results and their reproducibility unreliable. This paper describes a PCR-based method for easily identifying or confirming the species of origin of cell lines by using a panel of oligonucleotides specific for the nine animal species most common in cell culture laboratories. A panel of 35 human and animal cell lines, whose species of origin were previously confirmed by isoenzyme assay, was studied with nine species-specific primer pairs that specifically anneal to DNA sequences codifying for human, cat, dog, mouse, rat, horse, rabbit, African Green monkey cytochrome c oxidase subunit I (cox I), and one primer pair specific for the cytochrome b gene of Chinese hamster. The amplified fragments were analyzed by electrophoresis in ethidium bromide-stained 2% agarose gels. The method is simple, rapid, highly sensitive, and useful for routinely monitoring the species identity of cell cultures.     

Distinguishing biomechanical properties of intrinsic and extrinsic human wrist ligaments

Two intrinsic (scapholunate and lunotriquetral) and two extrinsic (radiolunate and radiocapitate) wrist ligaments were studied at high and low elongation rates (1 and 100 mm/min). Statistically significant differences among all four ligaments were noted for the viscoelastic and elastic components of stress versus strain for the fully recoverable strain and early permanent deformation stress for all ligaments. Intrinsic ligaments became permanently deformed at statistically significantly higher strain levels than the extrinsic ligaments and accept larger permanent deformation at strain levels below evident fiber failure. Ultimate strength data demonstrated significant rate dependency for stress and strain for all ligaments. Intrinsic ligaments failed statistically greater stress and strain levels than the extrinsic group. Some clinical implications of these findings are discussed.     

LAMP for human African trypanosomiasis: a comparative study of detection formats

Loop-mediated isothermal amplification (LAMP) is at the forefront of the search for innovative diagnostics for human African trypanosomiasis (HAT). Several simple endpoint detection methods have been developed for LAMP and here we compare four of these: (i) visualization of turbidity; (ii) addition of hydroxynaphthol blue before incubation; (iii) addition of calcein with MnCl₂ before incubation and (iv) addition of Quant-iT PicoGreen after incubation. These four methods were applied to four LAMP assays for the detection of human African trypanosomiasis, including two Trypanozoon specific and two Trypanosoma brucei rhodesiense specific reactions using DNA extracted from cryo-preserved procyclic form T. b. rhodesiense. A multi-observer study was performed to assess inter-observer reliability of two of these methods: hydroxynapthol blue and calcein with MnCl₂, using DNA prepared from blood samples stored on Whatman FTA cards. Results showed that hydroxynaphthol blue was the best of the compared methods for easy, inexpensive, accurate and reliable interpretation of LAMP assays for HAT. Hydroxynapthol blue generates a violet to sky blue colour change that was easy to see and was consistently interpreted by independent observers. Visible turbidity detection is not possible for all currently available HAT LAMP reactions; Quant-iT PicoGreen is expensive and addition of calcein with MnCl₂ adversely affects reaction sensitivity and was unpopular with several observers.     

The most reliable surface marker for the identification of colorectal cancer stem-like cells: A systematic review and meta-analysis

Several surface markers have been proposed for the identification and characterization of colorectal cancer stem-like cells (CR-CSLCs). However, their reliability in CR-CSLCs identification remains controversial. This study evaluated the correlation between all candidate surface marker's expression and CSLCs properties (tumorigenicity) through monitoring in vivo tumor incidence and final tumor volume. PubMed, Web of Science, and Scopus databases were systematically searched until November 2017. A total of 27 studies were found that met the inclusion criteria for cluster of differentiation 133 (CD133) and CD44 markers. Results indicated that either CD133 or CD44 positive cells caused about twofold increase in tumor volume compared with the negative cells (p < 0.05). In two groups of cells derived from primary tumors and cell lines, CD133 ⁺ cells had 25 and 1.45 times higher tumor incidence potential than CD133 ⁻ cells, respectively ( p < 0.05). Also, cohort evaluation showed that CD133 overexpression at protein level is a marker of poor overall survival in colorectal cancer (CRC) patients. While CD44 ⁺ cells displayed twofold tumorigenicity compared with the negative cells ( p < 0.05), combination of CD44 and CD133 showed about sevenfold tumorigenicity potential ( p < 0.05). In conclusion, the present meta-analysis suggests that CD133 is a robust biomarker to identify primary tumor CSLCs and can be proposed as a prognostic marker of CRC patient whereas it should be used with caution in cell lines. It seems to be more reliable to use CD133 in combination with CD44 as target biomarkers for the isolation of CR-CSLCs in both cell line and primary tumor cells populations.     

The pH dependent properties of metallotransferrins: a comparative study

The dependence on pH of the absorption and circular dichroic spectra of iron(III), cobalt(III) and copper(II) transferrins has been (re)investigated. In the alkaline region, the CD profiles of iron(III) and cobalt(III) transferrin are essentially pH independent up to pH 11; only for very high pH values (pH > 11) is breakdown of the cobalt(III) and iron(III) transferrin derivatives observed, without evidence of conformational rearrangements. By contrast, the CD profiles of copper transferrin show drastic changes in shape around pH 10; these spectral changes, which are fitted to a pK_a of ~10.4, are interpreted in terms of a substantial rearrangement of the local environment of the copper ions at high pH. Although the CD spectra of copper transferrin at alkaline pH strictly resemble those observed upon addition of modifier anions, the mechanism of site destabilization in the two cases is different; at variance with the case of modifier anions, our results suggest that the high pH form of copper transferrin still contains the synergistic anion. A¹³C NMR experiment has confirmed this view. In the acidic region, iron(III) and cobalt(III) transferrins are stable down to pH ~6. For lower pH values progressive metal detachment is observed without evidence of conformational changes; around pH 4.5 most bound metals are released. In the case of the less stable copper-transferrin, metal removal from the specific binding sites is already complete around pH 6.0; in concomitance with release from the primary sites, binding of copper ions to secondary sites is observed. Additional information has been gained from CD experiments in the far UV. The pH dependent properties of iron(III), cobalt(III) and copper(II) transferrin are discussed in the frame of the present knowledge of transferrin chemistry, particular emphasis being attributed to the comparison between tripositive and bipositive metal derivatives.     

An eye movement study for identification of suitable font characters for presentation on a computer screen

We are experiencing a shifting of media: from the printed paper to the computer screen. This transition is modifying the process of how we read and understand a text. It is very difficult to conclude on suitability of font characters based upon subjective evaluation method only. Present study evaluates the effect of font type on human cognitive workload during perception of individual alphabets on a computer screen. Twenty six young subjects volunteered for this study. Here, subjects have been shown individual characters of different font types and their eye movements have been recorded. A binocular eye movement recorder was used for eye movement recording. The results showed that different eye movement parameters such as pupil diameter, number of fixations, fixation duration were less for font type Verdana. The present study recommends the use of font type Verdana for presentation of individual alphabets on various electronic displays in order to reduce cognitive workload.     

Mechanical properties of the lateral collateral ligament: effect of cruciate instability in the rabbit

Concomitant soft tissue injury resulting from knee instability following cruciate rupture is a serious clinical problem. To study this injury mechanism, the biomechanical properties of the lateral collateral ligament were measured at 0, 4, 8, 12, and 16 weeks post-operatively in rabbits having the anterior and posterior cruciate ligaments sectioned. No significant changes were found in the ligament's cross-sectional area, tensile mechanical response, or in its hexosamine content. The predominant mode of ligament failure was by bone avulsion at the insertion sites (78 percent) with 86 percent of paired limbs failing in a similar manner.