Species-specific detection of Candida tropicalis using evolutionary conserved intein DNA sequences.

Inteins (internal proteins) are self-splicing transportable genetic elements present in conserved regions of housekeeping genes. The study highlights the importance of intein as a potential diagnostic marker for species-specific identification of Candida tropicalis, a rapidly emerging opportunistic human pathogen. Initial steps of primer validation, sequence alignment, phylogenetic tree analysis, gel electrophoresis and real-time polymerase chain reaction (PCR) assays were performed to confirm the specificity of the designed primers. The primers were selective for C. tropicalis with 100% inclusivity and showed no cross-species or cross-genera matches. The established technique is a prototype for developing multifaceted PCR assays and for point-of-care testing in near future. SIGNIFICANCE AND IMPACT OF THE STUDY: Development of molecular markers for specific detection of microbial pathogens using real-time polymerase chain reaction (PCR) is an appealing and challenging technique. A real-time PCR is an emerging technology frequently used to detect the aetiologic agents. In recent times, designing species-specific primers for pathogen detection is gaining momentum. The method offers rapid, accurate and cost-effective strategy to identify the target, thus providing sufficient time to instigate appropriate chemotherapy. The study highlights the use of intein DNA sequence as molecular markers for species-specific identification of Candida tropicalis. The study also offers a prototype model for developing multifaceted PCR assays using intein DNA sequences, and provides a developmental starting point for point-of-care testing in near future.

Effect of orally administered vibrio bacterin on immunity, survival and growth in tiger shrimp (Penaeus monodon) grow-out culture ponds.

Vibriosis is one of the important diseases causing economic loss to the shrimp industry worldwide. The present study reports field observations on the immune stimulatory effect of vibrio bacterin in commercial tiger shrimp (Penaeus monodon) grow-out culture ponds (n = 62) which were grouped under three stocking densities; low (6-8 nos per m2 ), medium (9-11 nos per m2 ) and high (12-14 nos per m2 ). The bacterin was administered in feed as a top dressing at final concentration equivalent to 2 × 108 CFU per kilogram feed twice a week throughout the culture period. In 20 representative ponds, total haemocyte count and prophenoloxidase activity in shrimp were significantly (P < 0·05) higher and anatomical deformities like, antennae cut (5·02 ± 2·42), tail rot (5·10 ± 1·74), rostrum cut (4·49 ± 2·19) and soft shell (10·05 ± 5·77) were significantly lower compared to controls in all the studied stocking densities. Significant (P < 0·05) improvement in production parameters like survival and production (kg ha1 ) was observed in all treatment ponds while similar improvement in average daily gain and feed conversion ratio could be observed in groups with low and medium stocking densities. Results of the study suggest that, oral administration of vibrio bacterin improves the immunity, reduces anatomical deformities and enhances the production in commercial shrimp culture operations. SIGNIFICANCE AND IMPACT OF THE STUDY: Administration of vibrio bacterin in feed as a top dressing induced immune stimulation as indicated by higher levels of total haemocyte count and prophenoloxidase. Further reduction in percentage of animals with anatomical deformities suggests the protection against subclinical bacterial infections. The overall improvement in the production parameters like, average daily gain, survival, feed conversation ratio and production in different shrimp stocking densities under commercial farming conditions suggested the possible development of an immune stimulant product based on the inactivated vibrio bacteria for improved health and production in Penaeus monodon shrimp farming.

A novel segmentation approach for noisy medical images using intuitionistic fuzzy divergence with neighbourhood-based membership function.

Medical image segmentation demands higher segmentation accuracy especially when the images are affected by noise. This paper proposes a novel technique to segment medical images efficiently using an intuitionistic fuzzy divergence-based thresholding. A neighbourhood-based membership function is defined here. The intuitionistic fuzzy divergence-based image thresholding technique using the neighbourhood-based membership functions yield lesser degradation of segmentation performance in noisy environment. Its ability in handling noisy images has been validated. The algorithm is independent of any parameter selection. Moreover, it provides robustness to both additive and multiplicative noise. The proposed scheme has been applied on three types of medical image datasets in order to establish its novelty and generality. The performance of the proposed algorithm has been compared with other standard algorithms viz. Otsu's method, fuzzy C-means clustering, and fuzzy divergence-based thresholding with respect to (1) noise-free images and (2) ground truth images labelled by experts/clinicians. Experiments show that the proposed methodology is effective, more accurate and efficient for segmenting noisy images.

Mechanistic modeling of vacuum UV advanced oxidation process in an annular photoreactor.

A novel mechanistic model that describes the vacuum UV advanced oxidation process in an annular photoreactor initiated by 172 nm and 185 nm (in combination with 253.7 nm, with and without exogenous H2O2) is presented in this paper. The model was developed from first principles by incorporating the vacuum UV-AOP kinetics into the theoretical framework of in-series continuous flow stirred tank reactors. After conducting a sensitivity analysis, model predictions were compared against experiments conducted under a variety of conditions: (a) photo-induced formation of hydrogen peroxide by water photolysis at 172 nm (for both air- and oxygen-saturated conditions); (b) photo-induced formation of hydrogen peroxide by water photolysis at 185 + 253.7 nm (in the presence of formic acid, with and without the initial addition of hydrogen peroxide); (c) direct photolysis of hydrogen peroxide by 253.7 nm; (d) degradation of formic acid by 185 + 253.7 nm (with and without initial addition of hydrogen peroxide); and (e) degradation of formic acid by 253.7 nm (with the addition of exogenous hydrogen peroxide). In all cases, the model was able to accurately predict the time-dependent profiles of hydrogen peroxide and formic acid concentrations. Two newly recognized aspects associated with water photolysis were identified through the use of the validated model. Firstly, unlike the 185 nm and 253.7 nm cases, water photolysis by the 172 nm wavelength revealed a depth of photoactive water layer an order of magnitude greater (∼230-390 µm, depending on the specific operating conditions) than the 1-log photon penetration layer (∼18 µm). To further investigate this potentially very important finding, a computational fluid dynamics model was set up to assess the role of transport mechanisms and species distributions within the photoreactor annulus. The model confirmed that short-lived hydroxyl radicals were present at a radial distance far beyond the ∼18 µm photon penetration layer. Secondly, kinetic simulations showed that the higher penetration depth of hydroxyl radicals was not caused by diffusive or convective transport phenomena but rather the effect of non-linear behavior of the complex reaction kinetics involved in the process.

[Landmark-based statistical procrustes analysis in the examination of breast shape and symmetry]. / Landmark-basierte statistische Procrustes Analyse bei der Untersuchung der Brustform und -symmetrie.

AIM: The aim of this presentation is the explanation of a mathematical analysis of a landmark-based procrustes calculation. The various components of breast symmetry and their impact on breast reconstruction have been examined. METHOD: In an objective breast shape analysis asymmetries are quantified as a measure of the difference between a 3D landmark configuration of a breast and its mirror image. 10 landmarks, 4 on each side and 2 for a definition of the midline were defined. For each landmark the 3D coordinates were calculated statistically with the help of the software Programme R. Over a midline from the jugulum to the manubrium sterni, a mirror image of the breast was constructed. The data for the breast to be reconstructed and the non-operated breast from 44 patients were investigated. The statistical differences of the Procrustes analysis were used for an asymmetry score and the proportions of the individual components were calculated. These included size, location and orientation of the breast as well as the individual inherent morphological surface form data of the breast. RESULTS: All 44 patients exhibited breast asymmetries and the mean asymmetry score amounted to 0.52. A calculation of the proportions of the individual components making up the asymmetry score revealed that morphological surface data made the largest contribution to the asymmetry score, closely followed by location, i. e., positioning of the breast on the thoracic wall. In contrast, the size of the breast and its orientation were of lesser relevance. CONCLUSION: When considering breast symmetry in plastic surgery, the form plays a more important role than the size as objectively calculated by the statistical procrustes analysis. Almost equally important as the form is the positioning of the breast on the thoracic wall which contributes significantly more to total breast symmetry than axial displacements.

Octaoctyl-substituted lutetium bisphthalocyanine for NADH biosensing.

Cyclic voltammetric and Raman and UV-vis spectroscopic measurements were performed on thin films of nonperipherally substituted bis[1,4,8,11,15,18,22,25-octakis(octyl)phthalocyaninato] lutetium(III) (R16LuPc2). Voltammograms exhibit one-electron quasi-reversible redox processes in 1.5 M LiClO4 aqueous solutions. The red-shift of the Q-band of R16LuPc2 in the UV-visible absorption spectra upon oxidation is attributed to the shortening of the inter-ring distance between the two phthalocyanine moieties. This observation is also consistent with the shift in the redox-sensitive vibrational modes in the Raman spectra due to the localization of the positive charge on phthalocyanine moieties. Neutralization of the oxidized R16LuPc2(+) film by dihydronicotinamide adenine dinucleotide (NADH) using different concentrations varying from 0.05 to 1 mM has been studied by UV-vis absorption and Raman spectroscopies. The reduction processes for a three month old film were found to be slower than those for freshly prepared films and showed a dependence upon NADH concentration. The data provide a basis for application of R16LuPc2 as a sensor for NADH.

Application of higher order cumulant features for cardiac health diagnosis using ECG signals.

Electrocardiogram (ECG) is the electrical activity of the heart indicated by P, Q-R-S and T wave. The minute changes in the amplitude and duration of ECG depicts a particular type of cardiac abnormality. It is very difficult to decipher the hidden information present in this nonlinear and nonstationary signal. An automatic diagnostic system that characterizes cardiac activities in ECG signals would provide more insight into these phenomena thereby revealing important clinical information. Various methods have been proposed to detect cardiac abnormalities in ECG recordings. Application of higher order spectra (HOS) features is a seemingly promising approach because it can capture the nonlinear and dynamic nature of the ECG signals. In this paper, we have automatically classified five types of beats using HOS features (higher order cumulants) using two different approaches. The five types of ECG beats are normal (N), right bundle branch block (RBBB), left bundle branch block (LBBB), atrial premature contraction (APC) and ventricular premature contraction (VPC). In the first approach, cumulant features of segmented ECG signal were used for classification; whereas in the second approach cumulants of discrete wavelet transform (DWT) coefficients were used as features for classifiers. In both approaches, the cumulant features were subjected to data reduction using principal component analysis (PCA) and classified using three layer feed-forward neural network (NN) and least square-support vector machine (LS-SVM) classifiers. In this study, we obtained the highest average accuracy of 94.52%, sensitivity of 98.61% and specificity of 98.41% using first approach with NN classifier. The developed system is ready clinically to run on large datasets.

Characterisation and identification of enzyme-producing autochthonous bacteria from the gastrointestinal tract of two Indian air-breathing fish.

Characterisation and identification of autochthonous enzyme-producing bacteria isolated from the proximal intestine and distal intestine of two species of Indian air-breathing fish, murrel (Channa punctatus) and stinging catfish (Heteropneustes fossilis), were investigated using conventional culture technique. Population levels of proteolytic strains were highest in the digestive tract of stinging catfish. In both species, the viable counts of amylase-producing bacteria were somewhat higher than cellulase-producing bacteria. Among the gut bacteria isolated, 8 strains (4 from murrel and 4 from stinging catfish) were selected as potent enzyme-producers on the basis of quantitative enzyme assays. All these strains were Gram-positive rods, but only four isolates (CPF4, CPH6, CPH7 and HFH4) were capable of forming endospores. The tested bacteria grew in wide range of temperatures and pH. The strains were further identified by 16S rRNA gene sequence analysis. Two strains, CPF3 (isolated from murrel) and HFH4 (isolated from stinging catfish) showed high similarity to Bacillus sp., strain HFH7 (isolated from the stinging catfish) was most closely related to Bacillus subtilis, while five strains belonged to Bacillus licheniformis. Based on the results of the present study, we suggest that incorporation of autochthonous enzyme-producing bacteria in aquafeeds merits further investigations.

Quantitative microscopy approach for shape-based erythrocytes characterization in anaemia.

Anaemia is one of the most common diseases in the world population. Primarily anaemia is identified based on haemoglobin level; and then microscopically examination of peripheral blood smear is required for characterizing and confirmation of anaemic stages. In conventional approach, experts visually characterize abnormality present in the erythrocytes under light microscope, and this evaluation process is subjective in nature and error prone. In this study, we have proposed a methodology using machine learning techniques for characterizing erythrocytes in anaemia associated with anaemia using microscopic images of peripheral blood smears. First, peripheral blood smear images are preprocessed based on grey world assumption technique and geometric mean filter for reducing unevenness of background illumination and noise reduction. Then erythrocyte cells are segmented using marker-controlled watershed segmentation technique. The erythrocytes in anaemia, such as, tear drop, echinocyte, acanthocyte, elliptocyte, sickle cells and normal erythrocytes cells have been characterized and classified based on their morphological changes. Optimal subset of features, ranked by information gain measure provides highest classification performance using logistic regression classifier in comparison with other standard classifiers.

In situ chemichromic studies of interactions between a lutetium bis-octaalkyl-substituted phthalocyanine and selected biological cofactors.

Spin-coated films, approximately 100 nm thick, of a newly synthesized bis[octakis(octyl)phthalocyaninato] lutetium(III) complex on ultrasonically cleaned glass substrates exhibit pronounced chemichromic behaviour with potential application in healthcare. In situ kinetic optical absorption spectroscopic measurements show that the phthalocyanine Q-band is red shifted by 60 nm upon oxidation arising from exposure to bromine vapour. Recovery to the original state is achieved by the treatment of the oxidized films with nicotinamide adenine dinucleotide and l-ascorbic acid (vitamin C) in an aqueous solution containing 1.5 M lithium perchlorate. The neutralization process is found to be governed by first-order kinetics. The linear increase of the reduction rate with increasing concentration of cofactors provides a basis for calibration of analyte concentrations ranging from 3.5 mM down to 0.03 mM.

Automated diagnosis of oral cancer using higher order spectra features and local binary pattern: a comparative study.

In the field of quantitative microscopy, textural information plays a significant role very often in tissue characterization and diagnosis, in addition to morphology and intensity. The objective of this work is to improve the classification accuracy based on textural features for the development of a computer assisted screening of oral sub-mucous fibrosis (OSF). In fact, the approach introduced is used to grade the histopathological tissue sections into normal, OSF without dysplasia (OSFWD) and OSF with dysplasia (OSFD), which would help the oral onco-pathologists to screen the subjects rapidly. The main objective of this work is to evaluate the use of Higher Order Spectra (HOS) features and Local Binary Pattern (LBP) features extracted from the epithelial layer in classifying normal, OSFWD and OSFD. For this purpose, we extracted twenty three HOS features and nine LBP features and fed them to a Support Vector Machine (SVM) for automated diagnosis. One hundred and fifty eight images (90 normal, 42 OSFWD and 26 OSFD images) were used for analysis. LBP features provide a good sensitivity of 82.85% and specificity of 87.84%, and the HOS features provide higher values of sensitivity (94.07%) and specificity (93.33%) using SVM classifier. The proposed system, can be used as an adjunct tool by the onco-pathologists to cross-check their diagnosis.

A comparative study of the effect of refining on charge of various pulps.

This work studied the influence of refining on various pulp properties such as freeness, specific surface area, specific volume, water retention value, surface charge, total charge and tensile index of five different pulps. At a freeness level the wheat straw pulp exhibited higher surface charge and total charge compared to other pulps. The specific surface area, specific volume and water retention value of the pulps increased with increased refining. The total fibre charge as determined by conductometric titrations was not affected by refining. However, the surface charge as determined by titrations with poly-DADMAC increased with refining. The increases in specific surface area of pulps by refining resulted in a higher fibre surface charge and also better fibre-fibre bonding. The linear regression models developed using experimental data were found to be accurate. The comparison of model predicted data and the experimental data showed an excellent agreement between them.

Characterisation of protein adsorption on different liquid crystal phthalocyaninethin films.

Bovine serum albumin (BSA) protein adsorption on thin spun films of different metal octakishexylthiophthalocyanine [(C(6)S)(8)PcM, M=Cu, Ni] derivatives is investigated by using three independent spectroscopic measurements namely Raman spectroscopy, ellipsometry and surface plasmon resonance imaging. Thermally induced molecular self-reorganisations in the phthalocyanine films are found to have produced the changes in the surface energy which, in turn, control protein adsorption. The amount of BSA adsorption on [(C(6)S)(8)PcNi] is more limited than that on [(C(6)S)(8)PcCu] and this observation is consistent with the results on the surface wettability obtained from the contact angle measurements. The shift from the plasmonic resonance wavelength because of the BSA adsorption was significantly larger for the heat-treated [(C(6)S)(8)PcCu] than as-deposited film. Similar measurements on the [(C(6)S)(8)PcNi] films showed a limited BSA adsorption. The results of surface plasmon resonance experiments are consistent with those obtained from Raman spectroscopic and ellipsometric techniques.

Surface plasmon resonance imaging for biosensing.

Surface plasmon resonance imaging (SPRI) is a useful tool for the study of surface biomolecular interactions allowing for label-free detection and elegant instrumentation. SPRI imaging system is described in this review with an emphasis on recent applications with examples of different biological interactions and high throughput analysis. Signal amplification in SPRI using nanoparticle and waveguide-based optical coupling is introduced. Finally the detection sensitivity of the SPRI system is examined in terms of other competitive methods.

Bilateral giant juvenile fibroadenoma of breasts.

An 11-year-old girl with rapidly enlarging bilateral breast lumps is reported. It was diagnosed as a case of juvenile fibroadenoma following fine needle aspiration cytology and confirmed on histopathological examination of the excised specimens.

Polaron hopping in some biomolecular solids and their charge transfer complexes.

The solid state spectroscopy of charge transfer complexes of biomolecules such as fatty acids, tripalmitin, lysozyme. folic acid, beta-carotene, cytochrome c, valinomycin and gramicidin has been carried out. The absorption coefficient is related with electronic conductivity. A half-power beta density is found common among these macromolecular solids, indicating photon-induced polaron hopping or hopping of a charge carrier between two branches of a polariton. Band gap vs full width at half-maximum of the mid-IR peak also reveals a linear relation.

Use of the electrochromic behaviour of lanthanide phthalocyanine films for nicotinamide adenine dinucleotide detection.

Electrochromic properties of spun films of bis[octakis(hexylthio)phthalocyaninato] dysprosium(III) were investigated for determining nicotinamide adenine dinucleotide hydride (NADH) in water solutions. A spin-coated film deposited on indium tin oxide electrode displays only one redox couple (at E1/2=0.78V). The films of [(C6H13S)8Pc]2Dy were modified chemically or electrochemically for the detection of reduced NADH in water solution. The modified film in the oxidized ([(C6H13S)8Pc]2Dy)+ form is believed to be reduced to its neutral form on interaction with NADH.

Structural and chemical changes in ultra-high-molecular-weight polyethylene due to gamma radiation-induced crosslinking and annealing in air.

Ultra-High-Molecular-Weight-Polyethylene (UHMWPE) is the material of choice for one of the articulating surfaces in many total joint replacements, notably hip and knee prostheses. The various methods used by the orthopaedic biomaterials industry to sterilize and anneal UHMWPE components, and the resulting oxidation and crosslinking, affect the mechanical wear resistance properties in ways still unknown at the microscopic and molecular levels. Transmission electron microscopy and chemical pyrolysis were used to quantify crosslinking induced by gamma irradiation and annealing in air. Changes in lamellar stacking and the amount of crosslinking suggest two types of crosslinking: relatively unstable crosslinks in the amorphous region initially resulting from gamma irradiation which are later replaced by more thermally stable crosslinks resulting from rearrangements at the annealing temperature. Lamellar mobility, the ability of crystalline lamellae to flow in the material, is enhanced during the transition from one type of bond to the other, and this appears to optimize near eight hours of annealing time. Results from decomposition and percent crystallinity measurements provide further support for this theory.

Zinc protoporphyrin IX attenuates closed head injury-induced edema formation, blood-brain barrier disruption, and serotonin levels in the rat.

The role of heme oxygenase (HO) in closed head injury (CHI) was examined using a potent HO and guanylyl cyclase inhibitor, zinc protoporphyrin (Zn-PP) in the rat. Blood-brain barrier (BBB) permeability to Evans blue and radioiodine, edema formation, and plasma and brain levels of serotonin were measured in control, CHI, and Zn-PP-treated CHI rats. CHI was produced by an impact of 0.224 N on the right parietal bone by dropping 114.6 g weight from a height of 20 cm in anesthetized rats. This concussive injury resulted in edema formation and brain swelling 5 hours after insult that was most pronounced in the contralateral hemisphere. The whole brain was edematous and remained in a semi-fluid state. Microvascular permeability disturbances to protein tracers were prominent in both cerebral hemispheres and the underlying cerebral structures. Plasma and brain serotonin showed pronounced increases and correlated with edema formation. Pretreatment with Zn-PP (10 mg/ kg, i.p) 30 minutes before or after CHI attenuated edema formation, brain swelling, plasma and brain serotonin levels, and microvascular permeability at 5 hours. Brain edema, BBB permeability, and serotonin levels were not attenuated when the compound was administered 60 minutes post-CHI suggesting that HO is involved in cellular and molecular mechanisms of edema formation and BBB breakdown early after CHI.