Butterfly eyespots evolved via cooption of an ancestral gene-regulatory network that also patterns antennae, legs, and wings.

Butterfly eyespots are beautiful novel traits with an unknown developmental origin. Here we show that eyespots likely originated via cooption of parts of an ancestral appendage gene-regulatory network (GRN) to novel locations on the wing. Using comparative transcriptome analysis, we show that eyespots cluster most closely with antennae, relative to multiple other tissues. Furthermore, three genes essential for eyespot development, Distal-less (Dll), spalt (sal), and Antennapedia (Antp), share similar regulatory connections as those observed in the antennal GRN. CRISPR knockout of cis-regulatory elements (CREs) for Dll and sal led to the loss of eyespots, antennae, legs, and also wings, demonstrating that these CREs are highly pleiotropic. We conclude that eyespots likely reused an ancient GRN for their development, a network also previously implicated in the development of antennae, legs, and wings.

Haemolymph transfusions transfer heritable learned novel odour preferences to naive larvae of Bicyclus anynana butterflies.

The mechanisms whereby environmental experiences of parents are transmitted to their offspring to impact their behaviour and fitness are poorly understood. Previously, we showed that naive Bicyclus anynana butterfly larvae, whose parents fed on a normal plant feed but coated with a novel odour, inherited an acquired preference towards that odour, which had initially elicited avoidance in the naive parents. Here, we performed simple haemolymph transfusions from odour-fed and control-fed larvae to naive larval recipients. We found that larvae injected with haemolymph from odour-fed donors stopped avoiding the novel odour, and their naive offspring preferred the odour more, compared to the offspring of larvae injected with control haemolymph. These results indicate that factors in the haemolymph, potentially the odour molecule itself, play an important role in odour learning and preference transmission across generations. Furthermore, this mechanism of odour preference inheritance, mediated by the haemolymph, bypasses the peripheral odour-sensing mechanisms taking place in the antennae, mouthparts or legs, and may mediate food plant switching and diversification in Lepidoptera or more broadly across insects.

ADGRU: Adaptive DenseNet with gated recurrent unit for automatic diagnosis of periodontal bone loss and stage periodontitis with tooth segmentation mechanism.

BACKGROUND: Periodontics and gingivitis are two of the most widely prevalent illnesses that affect people nowadays. The sixth most common disease in the world is periodontitis, and detecting periodontal bone loss is essential in the earlier condition and is crucial for the development of the proper diagnosis. Early bone loss detection can be assisted by using computer-assisted radiography examination. Understanding disease progression helps to select the most effective treatment action. OBJECTIVES: An effective deep model is suggested to detect periodontal bone loss at an earlier stage for preventing the progression of Periodontics bone loss. METHODS: This work is intimated by collecting images from online resources. Further, the images gathered from the dataset are preceded by the tooth segmentation which is done using DenseUNet + + . Further, the segmented images are given to the Adaptive DenseNet with Gated Recurrent Unit (AD-GRU) for detecting periodontal bone loss and this diagnosis is used for the periodontitis stage, where the ADGRU performance is augmented by optimizing the attributes using the Refined Red Kite Optimization Algorithm (RRKOA). RESULTS: The offered approach attained an accuracy of 94.45% which is higher than the88.63%, 90.58%, 89.54%, and 92.96% attained by the LSTM, DenseNet, GRU, DenseNet-GRU. DATA CONCLUSION: The findings of the simulation proved the designed framework outperformed the traditional model with high accuracy. CLINICAL RELEVANCE: The developed effectual deep model-based periodontal bone loss and stage periodontitis diagnosis structure is used in healthcare applications.

Predation favours Bicyclus anynana butterflies with fewer forewing eyespots.

There are fewer eyespots on the forewings versus hindwings of nymphalids but the reasons for this uneven distribution remain unclear. One possibility is that, in many butterflies, the hindwing covers part of the ventral forewing at rest and there are fewer forewing sectors to display eyespots (covered eyespots are not continuously visible and are less likely to be under positive selection). A second explanation is that having fewer forewing eyespots confers a selective advantage against predators. We analysed wing overlap at rest in 275 nymphalid species with eyespots and found that many have exposed forewing sectors without eyespots: i.e. wing overlap does not constrain the forewing from having the same number or more eyespots than the hindwing. We performed two predation experiments with mantids to compare the relative fitness of and attack damage patterns on two forms of Bicyclus anynana butterflies, both with seven hindwing eyespots, but with two (in wild-type) or four (in Spotty) ventral forewing eyespots. Spotty experienced more intense predation on the forewings, were shorter-lived and laid fewer eggs. These results suggest that predation pressure limits forewing eyespot number in B. anynana. This may occur if attacks on forewing eyespots have more detrimental consequences for flight than attacks on hindwing eyespots.

Successful management of severe hemolytic disease of the fetus due to anti-Jsb using intrauterine transfusions with serial maternal blood donations: a case report and a review of the literature.

BACKGROUND: The management of pregnant women with anti-Jsb is challenging due to the paucity of antigen-negative blood for fetal and neonatal transfusion. CASE REPORT: A 29-year-old woman with anti-Jsb was referred for assessment of recurrent fetal losses. With the presence of the sister as a historically matched donor, she was planned for active surveillance for fetal anemia during pregnancy. STUDY DESIGN AND METHODS: The fetus remained well until 21 weeks of gestation when signs of fetal anemia and early hydrops fetalis were noted. Anti-Jsb titer was at 128. The sister's red blood cells (RBCs) were cross-match incompatible. Urgent intrauterine transfusion (IUT) was performed with washed irradiated maternal RBCs, donated after cessation of heparin. The mother was given intravenous iron (IV-Fe) and continued on weekly recombinant human erythropoietin (rHu-EPO). RESULTS: Repeated IUTs were needed every 1 to 3 weeks. Throughout a 7-week period, three maternal donations were performed with total donated whole blood volume of 1250 mL, supporting four IUTs. At 29 weeks of gestation, the procedure was complicated by umbilical cord hematoma necessitating urgent cesarean section. A male newborn was delivered, transfused at birth, and subsequently treated with phototherapy and five top-up transfusions. CONCLUSION: This case represents a successful example of managing hemolytic disease of the fetus due to a rare antibody using maternal blood. It also supports previous data on safety of maternal donations during pregnancy and the use of combination of rHu-EPO and IV-Fe as a supportive measure.

Acquired preferences for a novel food odor do not become stronger or stable after multiple generations of odor feeding in Bicyclus anynana butterfly larvae.

Many herbivorous insects have specific host-plant preferences, and it is unclear how these preferences evolved. Previously, we found that Bicyclus anynana larvae can learn to prefer novel food odors from eating leaves with those odors and transmit those learned preferences to the next generation. It is uncertain whether such acquired odor preferences can increase across generations of repeated odor feeding and be maintained even in the absence of odor. In this study, we fed larvae with novel banana odor-coated leaves (odor-fed larvae) for five consecutive generations, without selection on behavioral choices, and measured how larval innate preferences changed over time. Then, we removed the odor stimulus from a larval subgroup, while the other group continued to be odor-fed. Our results show that larvae learned to prefer the novel odor within a generation of odor feeding and transmitted the learned preference to the next generation, as previously found. Odor-fed larvae preferred odor significantly more compared to control larvae across five generations of repeated odor or control feeding. However, this led neither to increased odor preference, nor its stabilization. This suggests that when butterfly larvae feed on a new host, a preference for that novel food plant may develop and be transmitted to the next generation, but this preference lasts for a single generation and disappears once the odor stimulus is removed.

Unusual domain architecture of aminoacyl tRNA synthetases and their paralogs from Leishmania major.

BACKGROUND: Leishmania major, a protozoan parasite, is the causative agent of cutaneous leishmaniasis. Due to the development of resistance against the currently available anti-leishmanial drugs, there is a growing need for specific inhibitors and novel drug targets. In this regards, aminoacyl tRNA synthetases, the linchpins of protein synthesis, have received recent attention among the kinetoplastid research community. This is the first comprehensive survey of the aminoacyl tRNA synthetases, their paralogs and other associated proteins from L. major. RESULTS: A total of 26 aminoacyl tRNA synthetases were identified using various computational and bioinformatics tools. Phylogenetic analysis and domain architectures of the L. major aminoacyl tRNA synthetases suggest a probable archaeal/eukaryotic origin. Presence of additional domains or N- or C-terminal extensions in 11 aminoacyl tRNA synthetases from L. major suggests possibilities such as additional tRNA binding or oligomerization or editing activity. Five freestanding editing domains were identified in L. major. Domain assignment revealed a novel asparagine tRNA synthetase paralog, asparagine synthetase A which has been so far reported from prokaryotes and archaea. CONCLUSIONS: A comprehensive bioinformatic analysis revealed 26 aminoacyl tRNA synthetases and five freestanding editing domains in L. major. Identification of two EMAP (endothelial monocyte-activating polypeptide) II-like proteins similar to human EMAP II-like proteins suggests their participation in multisynthetase complex formation. While the phylogeny of tRNA synthetases suggests a probable archaeal/eukaryotic origin, phylogeny of asparagine synthetase A strongly suggests a bacterial origin. The unique features identified in this work provide rationale for designing inhibitors against parasite aminoacyl tRNA synthetases and their paralogs.

Silver nanoparticles and magnetic beads with electrochemical measurement as a platform for immunosensing devices.

The simplicity and analytical utility of silver nanoparticles used as immunolabels with screen-printed measurement electrodes is illustrated by demonstrating an appropriate analytical signal for myoglobin (a protein marker for muscle damage) across a range of concentrations of physiological interest for distinguishing potential myocardial infarctions from normal background levels in serum. Silver nanoparticles were used as labels on one of a pair of anti-myoglobin clones while the other clone was covalently attached to magnetic beads. The two clones were selected so as to bind to different sites on the target protein and allow the formation of complexes containing both magnetic beads and silver nanoparticles. The magnetic beads enabled protein captured from test samples to be separated from other components, while the silver nanoparticles enabled the protein to be quantified. An oxidising potential, applied to screen-printed carbon electrodes, was used to dissolve silver without the need for an external oxidising agent. Silver ions released in the process were subsequently accumulated at the measurement electrodes by applying a suitable reducing potential and, finally, analytical signals were obtained by integrating the charges passed when accumulated silver was stripped from the electrodes by applying a potential ramp. The magnitudes of the measured charges were indicative of the concentrations of myoglobin in each of the test solutions.

Inheritance of Acquired Traits in Insects and Other Animals and the Epigenetic Mechanisms That Break the Weismann Barrier.

The credibility of the Weismann barrier has come into question. Several studies in various animal systems, from mice to worms, have shown that novel environmental stimuli can generate an altered developmental or behavioral trait that can be transmitted to offspring of the following generation. Recently, insects have become ideal models to study the inheritance of acquired traits. This is because insects can be reared in high numbers at low cost, they have short generation times and produce abundant offspring. Numerous studies have shown that an insect can modify its phenotype in response to a novel stimulus to aid its survival, and also that this modified phenotypic trait can be inherited by its offspring. Epigenetic mechanisms are likely at play but, most studies do not address the mechanisms that underlie the inheritance of acquired traits in insects. Here we first review general epigenetic mechanisms such as DNA methylation, histone acetylation and small noncoding RNAs that have been implicated in the transmission of acquired traits in animals, then we focus on the few insect studies in which these mechanisms have been investigated.

Transgenerational inheritance of learned preferences for novel host plant odors in Bicyclus anynana butterflies.

Many phytophagous insects have strong preferences for their host plants, which they recognize via odors, making it unclear how novel host preferences develop in the course of insect diversification. Insects may learn to prefer new host plants via exposure to their odors and pass this learned preference to their offspring. We tested this hypothesis by examining larval odor preferences before and after feeding them with leaves coated with control and novel odors and by examining odor preferences again in their offspring. Larvae of the parental generation developed a preference for two of these odors over their development. These odor preferences were also transmitted to the next generation. Offspring of butterflies fed on these new odors chose these odors more often than offspring of butterflies fed on control leaves. In addition, offspring of butterflies fed on banana odors had a significant naïve preference for the banana odors in contrast to the naïve preference for control leaves shown by individuals of the parental generation. Thus, butterflies can learn to prefer novel host plant odors via exposure to them during larval development and transmit these learned preferences to their offspring. This ability potentially facilitates shifts in host plant use over the course of insect diversification.

MulPSSM: a database of multiple position-specific scoring matrices of protein domain families.

Representation of multiple sequence alignments of protein families in terms of position-specific scoring matrices (PSSMs) is commonly used in the detection of remote homologues. A PSSM is generated with respect to one of the sequences involved in the multiple sequence alignment as a reference. We have shown recently that the use of multiple PSSMs corresponding to an alignment, with several sequences in the family used as reference, improves the sensitivity of the remote homology detection dramatically. MulPSSM contains PSSMs for a large number of sequence and structural families of protein domains with multiple PSSMs for every family. The approach involves use of a clustering algorithm to identify most distinct sequences corresponding to a family. With each one of the distinct sequences as reference, multiple PSSMs have been generated. The current release of MulPSSM contains approximately 33,000 and approximately 38,000 PSSMs corresponding to 7868 sequence and 2625 structural families. A RPS_BLAST interface allows sequence search against PSSMs of sequence or structural families or both. An analysis interface allows display and convenient navigation of alignments and domain hits. MulPSSM can be accessed at http://crick.mbu.iisc.ernet.in/~mulpssm.

Automated Detection and Classification of Microcalcification Clusters with Enhanced Preprocessing and Fractal Analysis

This paper addresses the automated detection of microcalcification clusters from mammogram images by enhanced preprocessing operations on digital mammograms for automated extraction of breast tissue from background, removing artefacts occurring during image registration using X-rays, followed by fractal analysis of suspicious regions. Identification of breast of either left or right and realigning them to a standard position forms a primitive step in preprocessing of mammograms. As the next step in the process, pectoral muscles are separated. Suspicious regions of microcalcifications are identified and are subjected to further analysis of classifying it as benign or malignant. Texture features are representative of its malignancy and fractal analysis was carried out on extracted suspicious regions for its texture features. Principal Component Analysis was carried out to extract optimal features. Ten features were found to be an optimal number of reduced texture features without compromising on classification accuracy. Scaled conjugate Gradient Back propagation network was used for classification using reduced texture features obtained from PCA analysis. By varying hidden layer neurons, accuracy of results achieved by proposed methods is analysed and is calculated to reach maximum accuracy with an optimal level of 15 neurons. Accuracy of 96.3% was achieved with 10 fractal features as input to neural network and 15 hidden layer neurons in neural network designed. The design of architecture is finalised with maximised accuracy for labelling microcalcification clusters as benign or malignant.

Strategies for the effective identification of remotely related sequences in multiple PSSM search approach.

Searches using position specific scoring matrices (PSSMs) have been commonly used in remote homology detection procedures such as PSI-BLAST and RPS-BLAST. A PSSM is generated typically using one of the sequences of a family as the reference sequence. In the case of PSI-BLAST searches the reference sequence is same as the query. Recently we have shown that searches against the database of multiple family-profiles, with each one of the members of the family used as a reference sequence, are more effective than searches against the classical database of single family-profiles. Despite relatively a better overall performance when compared with common sequence-profile matching procedures, searches against the multiple family-profiles database result in a few false positives and false negatives. Here we show that profile length and divergence of sequences used in the construction of a PSSM have major influence on the performance of multiple profile based search approach. We also identify that a simple parameter defined by the number of PSSMs corresponding to a family that is hit, for a query, divided by the total number of PSSMs in the family can distinguish effectively the true positives from the false positives in the multiple profiles search approach.

Cotyledonoid leiomyoma in pregnancy.

BACKGROUND: Benign smooth-muscle tumors (leiomyomata) are the most frequent tumors found in the female genital tract. They are easy to diagnose in pregnancy and usually managed conservatively. Some variant forms with unusual infiltrative growth pattern have been known, but they are rare in pregnancy. The variant forms pose diagnostic and management difficulties. CASE: An unusual type of leiomyoma (cotyledonoid), adhering to the bowels and occupying the whole left broad ligament, was excised completely at 14 weeks of gestation, and the pregnancy continued to term. CONCLUSION: Familiarity with benign uterine smooth-muscle tumors with unusual growth patterns by gynecologists and pathologists is essential in avoiding over-treatment.

Analysis on sliding helices and strands in protein structural comparisons: a case study with protein kinases.

Protein structural alignments are generally considered as 'golden standard' for the alignment at the level of amino acid residues. In this study we have compared the quality of pairwise and multiple structural alignments of about 5900 homologous proteins from 718 families of known 3-D structures. We observe shifts in the alignment of regular secondary structural elements (helices and strands) between pairwise and multiple structural alignments. The differences between pairwise and multiple structural alignments within helical and beta-strand regions often correspond to 4 and 2 residue positions respectively. Such shifts correspond approximately to "one turn" of these regular secondary structures. We have performed manual analysis explicitly on the family of protein kinases. We note shifts of one or two turns in helix-helix alignments obtained using pairwise and multiple structural alignments. Investigations on the quality of the equivalent helix-helix, strand-strand pairs in terms of their residue side-chain accessibilities have been made. Our results indicate that the quality of the pairwise alignments is comparable to that of the multiple structural alignments and, in fact, is often better. We propose that pairwise alignment of protein structures should also be used in formulation of methods for structure prediction and evolutionary analysis.

Role of day 4 HCG as an early predictor of success after methotrexate therapy for ectopic pregnancies.

OBJECTIVE: To validate whether change in serum HCG levels between days 0 and 4 confer any prognostic value during methotrexate therapy and to quantify its change. STUDY DESIGN: This is a retrospective study of 48 tubal ectopic pregnancies treated with single dose methotrexate protocol at University Hospital, Muscat, Oman from January 2012 to December 2013. The clinical outcome was analyzed based on the complete resolution of HCG levels or need for additional doses of methotrexate or recourse to surgery. The percentage change in HCG levels between days 0 and 4 (HCG index) of methotrexate were calculated and receiver operator characteristics curve was plotted to identify the best cutoff levels. In order to get a robust 95% confidence interval, bootstrap method using R software was done using 1000 re-sampling. ROC curve and the predictive values were estimated using MEDCALC software. RESULTS: The mean HCG level on day 4 is significantly higher in treatment failure group (4254±4095 IU/L vs. 2109±3646 IU/L, P=0.008). The HCG levels between day 0 and 4 decreased in 42.7% (21/48) of cases and 80.9% of these cases had treatment success. The HCG levels increased in 57.4% (27/48) of cases and 33.3% of these cases had treatment success. (P=0.001). A 10 percent decline in day 4 HCG levels predict the treatment success with sensitivity of 77% and Specificity 81%. The area under the ROC curve was 0.82 (95% CI: 0.67-0.92), (P<0.001). CONCLUSION: The success with single dose of methotrexate therapy for tubal ectopic pregnancies was predicted early in the course of treatment by following three key findings: the absolute mean HCG values on day 4, decrease in HCG level from day 0 to 4 and 10% or more fall in day 4 HCG levels.

Identification of patients with preeclampsia from normal subjects using wavelet-based spectral analysis of heart rate variability.

BACKGROUND: The spectral analysis of the heart rate variability (HRV) shows a decrease in the power of the high frequency (HF) component in preeclamptic pregnancy compared with normal pregnancy; such a decrease is associated with an increase in the low frequency (LF) and the very low frequency (VLF) power. The physiological interpretation is that preeclamptic pregnancy is associated with a facilitation of sympathetic regulation and an attenuation of parasympathetic influence of HR compared with non-pregnancy and normal pregnancy. OBJECTIVE: To use an efficient nased on spectral analysis non-invasive technique to identify preeclamptic pregnant subjects from normal pregnant in Oman. METHODS: The soft-decision wavelet-based technique is implemented to find the power of the HRV bands in high resolution manner compared to the classical fast Fourier Transform method. Data was obtained from 20 preeclamptic pregnant subjects and 20 normal pregnant controls of the same pregnancy duration, obtained from Nizwa and Sultan Qaboos University hospitals in Oman. RESULTS: The soft-decision wavelet method succeeds to identify patients from normal pregnant with specificity, sensitivity and accuracy of 90%, 80% and 85%, respectively, compared to the FFT which results in 75% specificity, sensitivity and accuracy. CONCLUSION: The LF power obtained by Soft-decision wavelet decomposition is shown to be a successful feature for identification of preeclampsia.

Integration of related sequences with protein three-dimensional structural families in an updated version of PALI database.

The database of Phylogeny and ALIgnment of homologous protein structures (PALI) contains three-dimensional (3-D) structure-dependent sequence alignments as well as structure-based phylogenetic trees of protein domains in various families. The latest updated version (Release 2.1) comprises of 844 families of homologous proteins involving 3863 protein domain structures with each of these families having at least two members. Each member in a family has been structurally aligned with every other member in the same family using two proteins at a time. In addition, an alignment of multiple structures has also been performed using all the members in a family. Every family with at least three members is associated with two dendrograms, one based on a structural dissimilarity metric and the other based on similarity of topologically equivalenced residues for every pairwise alignment. Apart from these multi-member families, there are 817 single member families in the updated version of PALI. A new feature in the current release of PALI is the integration, with 3-D structural families, of sequences of homologues from the sequence databases. Alignments between homologous proteins of known 3-D structure and those without an experimentally derived structure are also provided for every family in the enhanced version of PALI. The database with several web interfaced utilities can be accessed at: http://pauling.mbu.iisc.ernet.in/~pali.

SUPFAM: a database of sequence superfamilies of protein domains.

BACKGROUND: SUPFAM database is a compilation of superfamily relationships between protein domain families of either known or unknown 3-D structure. In SUPFAM, sequence families from Pfam and structural families from SCOP are associated, using profile matching, to result in sequence superfamilies of known structure. Subsequently all-against-all family profile matches are made to deduce a list of new potential superfamilies of yet unknown structure. DESCRIPTION: The current version of SUPFAM (release 1.4) corresponds to significant enhancements and major developments compared to the earlier and basic version. In the present version we have used RPS-BLAST, which is robust and sensitive, for profile matching. The reliability of connections between protein families is ensured better than before by use of benchmarked criteria involving strict e-value cut-off and a minimal alignment length condition. An e-value based indication of reliability of connections is now presented in the database. Web access to a RPS-BLAST-based tool to associate a query sequence to one of the family profiles in SUPFAM is available with the current release. In terms of the scientific content the present release of SUPFAM is entirely reorganized with the use of 6190 Pfam families and 2317 structural families derived from SCOP. Due to a steep increase in the number of sequence and structural families used in SUPFAM the details of scientific content in the present release are almost entirely complementary to previous basic version. Of the 2286 families, we could relate 245 Pfam families with apparently no structural information to families of known 3-D structures, thus resulting in the identification of new families in the existing superfamilies. Using the profiles of 3904 Pfam families of yet unknown structure, an all-against-all comparison involving sequence-profile match resulted in clustering of 96 Pfam families into 39 new potential superfamilies. CONCLUSION: SUPFAM presents many non-trivial superfamily relationships of sequence families involved in a variety of functions and hence the information content is of interest to a wide scientific community. The grouping of related proteins without a known structure in SUPFAM is useful in identifying priority targets for structural genomics initiatives and in the assignment of putative functions. Database URL: http://pauling.mbu.iisc.ernet.in/~supfam.

Uterine inversion and corpus malignancies: a historical review.

Inversion of the uterus is a rare clinical problem. It is encountered as an obstetric emergency and a diagnostic challenge in gynecology. Association of uterine inversion with malignancies such as endometrial carcinoma or sarcoma is even rarer. Accurate diagnosis is essential before treatment, so that appropriate management is offered. A history of uterine inversion and a review of literature is presented especially of its association with corpus malignancies.