Small nerve fiber quantification in the diagnosis of diabetic sensorimotor polyneuropathy: Comparing corneal confocal microscopy with intraepidermal nerve fiber density

OBJECTIVE Quantitative assessment of small fiber damage is key to the early diagnosis and assessment of progression or regression of diabetic sensorimotor polyneuropathy (DSPN). Intraepidermal nerve fiber density (IENFD) is the current gold standard, but corneal confocal microscopy (CCM), an in vivo ophthalmic imaging modality, has the potential to be a noninvasive and objective image biomarker for identifying small fiber damage. The purpose of this study was to determine the diagnostic performance of CCM and IENFD by using the current guidelines as the reference standard. RESEARCH DESIGN AND METHODS Eighty-nine subjects (26 control subjects and 63 patients with type 1 diabetes), with and without DSPN, underwent a detailed assessment of neuropathy, including CCM and skin biopsy. RESULTS Manual and automated corneal nerve fiber density (CNFD) (P < 0.0001), branch density (CNBD) (P < 0.0001) and length (CNFL) (P < 0.0001), and IENFD (P < 0.001) were significantly reduced in patients with diabetes with DSPN compared with control subjects. The area under the receiver operating characteristic curve for identifying DSPN was 0.82 for manual CNFD, 0.80 for automated CNFD, and 0.66 for IENFD, which did not differ significantly (P = 0.14). CONCLUSIONS This study shows comparable diagnostic efficiency between CCM and IENFD, providing further support for the clinical utility of CCM as a surrogate end point for DSPN.

A chitotetrose specific lectin from Luffa acutangula :Physico-chemical properties and the assignment of orientation of sugars in the lectin binding site

A chitooligosaccharide specific lectin (Luffa acutangula agglutinin) has been purified from the exudate of ridge gourd fruits by affinity chromatography on soybean agglutininglycopeptides coupled to Sepharose-6B. The affinity purified lectin was found homogeneous by polyacrylamide gel electrophoresis, in sodium dodecyl sulphate-polyacrylamide gels, by gel filtration on Sephadex G-100 and by sedimentation velocity experiments. The relative molecular weight of this lectin is determined to be 48,000 ± 1,000 by gel chromatography and sedimentation equilibrium experiments. The sedimentation coefficient (S20, w) was obtained to be 4·06 S. The Stokes’ radius of the protein was found to be 2·9 nm by gel filtration. In sodium dodecyl sulphate-polyacrylamide gel electrophoresis the lectin gave a molecular weight of 24,000 in the presence as well as absence of 2-mercaptoethanol. The subunits in this dimeric lectin are therefore held by non-covalent interactions alone. The lectin is not a glycoprotein and circular dichroism spectral studies indicate that this lectin has 31% α-helix and no ß-sheet. The lectin is found to bind specifically to chitooligosaccharides and the affinity of the lectin increases with increasing oligosaccharide chain length as monitored by near ultra-violetcircular dichroism and intrinsic fluorescence titration. The values of ΔG, ΔΗ and ΔS for the binding process showed a pronounced dependence on the size of the oligosaccharide. The values for both ΔΗ and ΔS show a significant increase with increase in the oligosaccharide chain length showing that the binding of higher oligomers is progressively more favoured thermodynamically than chitobiose itself. The thermodynamic data is consistent with an extended binding site in the lectin which accommodates a tetrasaccharide. Based on the thermodynamic data, blue shifts and fluorescence enhancement, spatial orientation of chitooligosaccharides in the combining site of the lectin is assigned.

On the orientation of grain boundaries in cadmium recrystallized during hot deformatio

The authors are grateful to Professor K. P. Abraham for the provision of facilities and encouragement. One of us (PRR) acknowledges the award of a National Associateship by the UGC which facilitated a short-time visit to the Indian Institute of Science.

A hypothesis on the role of hydroxyproline in stabilizing collagen structure

The possibility of hydroxyproline residues stabilizing the collagen triple-helical structure by the formation of additional hydrogen bonds through their γ-hydroxyl group has been studied from structural considerations. It is not possible for this hydroxyl group to form a direct hydrogen bond with a suitable group in a neighbouring chain of the triple-helical protofibril. However, in the modified one-bonded structure, which is stabilized by additional hydrogen bonds being formed through water molecules as intermediaries (put forward in 1968 by Ramachandran, G. N. and Chandrasekharan, R.), it is found that the γ-hydroxyl group of hydroxyproline can form a good hydrogen bond with the water oxygen as acceptor, the hydrogen bond length being 2.82 Å. It is proposed that, in addition to stabilizing the collagen triple-helical structure due to the stereochemical properties of the pyrrolidine ring, hydroxyproline gives added stability by the formation of an extra hydrogen bond. Experimental studies on the determination of shrinkage and denaturation temperatures of native collagen and its synthetic analogues, as a function of their hydroxyproline content, are being undertaken to test this hypothesis.

Stabilization of the collagen structure by hydroxyproline residues

The molecular structure of collagen is now accepted to be based on a triple-stranded coiled-coil, in which the three strands are held together predominantly by hydrogen bonds. Recent experimental evidence has shown that the presence of hydroxyproline residues in the third position of the repeating tripeptide unit lends additional stability to the collagen structure. In this paper, we report a model structure, which is supported by these observations. In a model structure proposed earlier, there are two hydrogen bonds per tripeptide unit, one of which is a direct interchain hydrogen bond, while the second hydrogen bond can be formedvia a water molecule. It has now been shown that the same water molecule can also form a hydrogen bond with the oxygen of theγ-hydroxyl group of hydroxyproline in the third position in the sequence (Gly-R2-R3). This hydroxyl group can also take part in an inter-triple-helix hydrogen bond. Our studies thus show the role played by hydroxyproline residues in the structure and stability of collagen.

Average Orientation of Molecules Dissolved in Nematic Liquid Crystals of Opposite Diamagnetic Anisotropies

ZLI-1167 is a ternary mixture of nematic liquid crystals with negative diamagnetic anisotropy. It has, therefore, been used as a solvent where the spinning of the samples around the vertical axis in the conventional electromagnets without destroying the orientation of the dissolved molecules is possible in NMR experiments. This results in sharp lines with widths up to 1 Hz in the spectra.1,2 In an NMR system using a superconducting magnet (where the magnetic field direction is along the axis of spinning of the sample), it is possible to use even the nematic liquid crystals with positive diamagnetic anisotropy such as N-(p'-methoxybenzylidene)-p-n-butylaniline (MBBA) or N-(p'-ethoxybenzylidene)-p-n-butylaniline (EBBA) to obtain the spectra with sample spinning with equally sharp lines.3 The orientational behaviour of the dissolved molecules as a function of relative concentrations of the two solvents is investigated and the results are reported in the present communication.

Steroechemistry of the Pyrrolidine rings in the collagen structure

In the collagen triple-helical structure, large side groups occuring at location 3 in the repeating triplet sequences (Gly-Rz-Rz)n are appreciably constrained if a proline residue occurs as Rz in a neighbouring chain. The severity of the steric hindrance depends on the geometry of the prolyl ring. In this paper we propose two different puckerir.gs for the proline ring, the first one being energetically favorable for most types of residue sequences commonly found in collegen while the second is preferable when an amino acid residue with a large side group occurs at location 3 in a neighbouring chain. The puckering of the pyrrolidine ring of hydroxyproline, as proposed earlier, is quite favorable from energy as well as stereochemical considerations.

Fourier transform of the collagen triple-helical structure and its significance

The Fourier transforms of the collagen molecular structure have been calculated taking into consideration various side chain atoms, as well as the presence of bound water molecules. There is no significant change in the calculated intensity distribution on including the side chain atoms of non-imino-acid residues. Taking into account the presence of about two bound water molecules per tripeptide unit, the agreement with the observed x-ray pattern is slightly improved. Fourier transforms have also been calculated for the detailed molecular geometries proposed from other laboratories. It is found that there are no major differences between them, as compared to our structure, either in the positions of peak intensity or in the intensity distribution. Hence it is not possible to judge the relative merits of the various molecular geometries for the collagen triple helix from a comparison of the calculated transforms with the meagre data available from its x-ray fibre pattern. It is also concluded that the collagen molecular structure should be regarded as a somewhat flexible chain structure, capable of adapting itself to the requirements of the different side groups which occur in each local region.

A Novel Opto Electro Mechanical Interrogation System for Measuring Wavelength Shifts in Fiber Bragg Grating (FBG) Sensors

Here, we describe a novel FBG interrogation system in which FBGs are used as both sensing and reference elements. The reference FBGs is bonded to a mechanical flexure system having a linear amplification of 1:3.5, which is actuated using a piezo-actuator by applying a 0-150V ramp. The lengths of the reference gratings decide the maximum strain that can be applied to the reference grating, which in turn decides that strain range which can be interrogated. The main advantages of the present system are the on-line measurement of the wavelength shifts, small size, good sensitivity, multiplexing capability and low cost.

Why and when do learning goal orientation and attitude decrease with aging? The role of perceived remaining time and work centrality

We conducted two studies to improve our understanding of why and when older workers are focused on learning. Based on socioemotional selectivity theory, which proposes that goal focus changes with age and the perception of time, we hypothesized and found that older workers perceive their remaining time at work as more limited than younger workers which, in turn, is associated with lower learning goal orientation and a less positive attitude toward learning and development. Furthermore, we hypothesized and found that high work centrality buffers the negative association between age and perceived remaining time, and thus the indirect negative effects of age on learning goal orientation and attitude toward learning and development (through perceived remaining time). These findings suggest that scholars and practitioners should take workers’ perceived remaining time and work centrality into account when examining or stimulating learning activities among aging workers.

Compact fiber Bragg grating dynamic strain sensor cum broadband thermometer for thermally unstable ambience

An instrument for simultaneous measurement of dynamic strain and temperature in a thermally unstable ambience has been proposed, based on fiber Bragg grating technology. The instrument can function as a compact and stand-alone broadband thermometer and a dynamic strain gauge. It employs a source wavelength tracking procedure for linear dependence of the output on the measurand, offering high dynamic range. Two schemes have been demonstrated with their relative merits. As a thermometer, the present instrumental configuration can offer a linear response in excess of 500 degrees C that can be easily extended by adding a suitable grating and source without any alteration in the procedure. Temperature sensitivity is about 0.06 degrees C for a bandwidth of 1 Hz. For the current grating, the upper limit of strain measurement is about 150 mu epsilon with a sensitivity of about 80 n epsilon Hz(-1/2). The major source of uncertainty associated with dynamic strain measurement is the laser source intensity noise, which is of broad spectral band. A low noise source device or the use of optical power regulators can offer improved performance. The total harmonic distortion is less than 0.5% up to about 50 mu epsilon, 1.2% at 100 mu epsilon and about 2.3% at 150 mu epsilon. Calibrated results of temperature and strain measurement with the instrument have been presented. Traces of ultrasound signals recorded by the system at 200 kHz, in an ambience of 100-200 degrees C temperature fluctuation, have been included. Also, the vibration spectrum and engine temperature of a running internal combustion engine has been recorded as a realistic application of the system.

Inhibition of Tyrosine Phosphorylation of Sperm Flagellar Proteins, Outer Dense Fiber Protein-2 and Tektin-2, Is Associated With Impaired Motility During Capacitation of Hamster Spermatozoa

In mammals, acquisition of fertilization competence of spermatozoa is dependent on the phenomenon of sperm capacitation. One of the critical molecular events of sperm capacitation is protein tyrosine phosphorylation. In a previous study, we demonstrated that a specific epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitor, tyrphostin-A47, inhibited hamster sperm capacitation, accompanied by a reduced sperm protein tyrosine phosphorylation. Interestingly, a high percentage of tyrphostin-A47-treated spermatozoa exhibited circular motility, which was associated with a distinct hypo-tyrosine phosphorylation of flagellar proteins, predominantly of Mr 45,000-60,000. In this study, we provide evidence on the localization of capacitation-associated tyrosine-phosphorylated proteins to the nonmembranous, structural components of the sperm flagellum. Consistent with this, we show their ultrastructural localization in the outer dense fiber, axoneme, and fibrous sheath of spermatozoa. Among hypo-tyrosine phosphorylated major proteins of tyrphostin-A47-treated spermatozoa, we identified the 45 kDa protein as outer dense fiber protein-2 and the 51 kDa protein as tektin-2, components of the sperm outer dense fiber and axoneme, respectively. This study shows functional association of hypo-tyrosine-phosphorylation status of outer dense fiber protein-2 and tektin-2 with impaired flagellar bending of spermatozoa, following inhibition of EGFR-tyrosine kinase, thereby showing the critical importance of flagellar protein tyrosine phosphorylation during capacitation and hyperactivation of hamster spermatozoa.