Transcriptome sequencing of the endangered land snail Karaftohelix adamsi from the Island Ulleung: De novo assembly, annotation, valuation of fitness genes and SSR markers.

BACKGROUND: The Bradybaenidae snail Karaftohelix adamsi is endemic to Korea, with the species tracked from Island Ulleung in North Gyeongsang Province of South Korea. K. adamsi has been classified under the Endangered Wildlife Class II species of Korea and poses a severe risk of extinction following habitat disturbances. With no available information at the DNA (genome) or mRNA (transcriptome) level for the species, conservation by utilizing informed molecular resources seems difficult. OBJECTIVE: In this study, we used the Illumina short-read sequencing and Trinity de novo assembly to draft the reference transcriptome of K. adamsi. RESULTS: After assembly, 13,753 unigenes were obtained of which 10,511 were annotated to public databases (a maximum of 10,165 unigenes found homologs in PANM DB). A total of 6,351, 3,535, 358, and 3,407 unigenes were ascribed to the functional categories under KOG, GO, KEGG, and IPS, respectively. The transcripts such as the HSP 70, aquaporin, TLR, and MAPK, among others, were screened as putative functional resources for adaptation. DNA transposons were found to be thickly populated in comparison to retrotransposons in the assembled unigenes. Further, 2,164 SSRs were screened with the promiscuous presence of dinucleotide repeats such as AC/GT and AG/CT. CONCLUSION: The transcriptome-guided discovery of molecular resources in K. adamsi will not only serve as a basis for functional genomics studies but also provide sustainable tools to be utilized for the protection of the species in the wild. Moreover, the development of polymorphic SSRs is valuable for the identification of species from newer habitats and cross-species genotyping.

A review of the endangered mollusks transcriptome under the threatened species initiative of Korea.

Transcriptome studies for conservation of endangered mollusks is a proactive approach towards managing threats and uncertainties facing these species in natural environments. The population of these species is declining due to habitat destruction, illicit wildlife trade, and global climate change. These activities risk the free movement of species across the wild landscape, loss of breeding grounds, and restrictions in displaying the physiological attributes so crucial for faunal welfare. Gastropods face the most negative ecological effects and have been enlisted under Korea's protective species consortium based on their population dynamics in the last few years. Moreover, with the genetic resources restricted for such species, conservation by informed planning is not possible. This review provides insights into the activities under the threatened species initiative of Korea with special reference to the transcriptome assemblies of endangered mollusks. The gastropods such as Ellobium chinense, Aegista chejuensis, Aegista quelpartensis, Incilaria fruhstorferi, Koreanohadra kurodana, Satsuma myomphala, and Clithon retropictus have been represented. Moreover, the transcriptome summary of bivalve Cristaria plicata and Caenogastropoda Charonia lampas sauliae is also discussed. Sequencing, de novo assembly, and annotation identified transcripts or homologs for the species and, based on an understanding of the biochemical and molecular pathways, were ascribed to predictive gene function. Mining for simple sequence repeats from the transcriptome have successfully assisted genetic polymorphism studies. A comparison of the transcriptome scheme of Korean endangered mollusks with the genomic resources of other endangered mollusks have been discussed with homologies and analogies for dictating future research.

Time-Sequencing of the Neutrophil-to-Lymphocyte Ratio to Predict Prognosis of Triple-Negative Breast Cancer.

Since triple-negative breast cancers (TNBCs) have varying prognoses, it is important to identify subgroups with particularly poor prognosis. The aim of this study was to assess whether changes in the neutrophil-to-lymphocyte ratio (NLR) during the treatment process were associated with poor prognosis in TNBC patients. This study included 600 TNBC patients who underwent surgery from January 2005 to December 2016. The associations of the NLR and clinicopathologic factors with breast cancer recurrence and survival in patients who underwent both definitive local treatment (total mastectomy or breast-conserving surgery with radiotherapy) and systemic chemotherapy were analyzed. The NLRs at four time points (before surgery, before chemotherapy, before radiotherapy, and 1 year after surgery) were assessed. The univariate analysis showed that changes in the NLR before the start of radiotherapy (odds ratio: 1.115, confidence interval: 1.011-1.229) and 1 year after surgery (odds ratio: 1.196, confidence interval: 1.057-1.354) significantly increased the risk of recurrence or death. In multivariate analysis, T stage, N stage, and changes in the NLR were significant factors. A time-sequenced NLR may reflect the prognosis of TNBC patients. A poor prognosis is expected in patients whose NLR increases during treatment compared to the preoperative NLR, and additional treatment is needed.

De novo transcriptome sequencing of triton shell Charonia lampas sauliae: Identification of genes related to neurotoxins and discovery of genetic markers.

Charonia lampas sauliae (triton snails, triton shells or tritons; Mollusca, Caenogastropoda, Littorinimorpha, Ranellidae) is a marine species with a wide distribution. In Korea, this species is listed as vulnerable and is regionally protected as an endangered species. Here, we report the first comprehensive transcriptome dataset of C. lampas sauliae obtained using the Illumina HiSeq 2500 platform. In total, 97.68% of raw read sequences were processed as clean reads. Of the 577,478 contigs obtained, 146,026 sequences were predicted to contain coding regions. About 89.34% of all annotated unigene sequences showed homologous matches to protein sequences in PANM DB (Protostome database). Further, about one-third of the unigene sequences were annotated using the UniGene, Swiss-Prot, Clusters of Orthologous Groups (COG) and Gene Ontology (GO) databases. In total, 190 enzymes were predicted under key metabolic pathways under stood through Kyoto Encyclopedia of Genes and Genomes (KEGG) database annotation. Repetitive elements such as long terminal repeats (LTRs), short interspersed nuclear elements (SINEs), long interspersed nuclear elements (LINEs), and DNA elements were enriched in the unigene sequences. Among the identified transcripts were the channel proteins, some of which were blocked by tetrodotoxin, which is thought to be synthesized by symbiotic bacteria inhabiting the shells. In addition, conotoxin superfamily peptides, such as B-conotoxin, conotoxin superfamily T and alpha-conotoxin, were identified, which may have relevance to biomedical and evolutionary research. A transcriptome-wide search for polymorphic loci identified 21,568 simple sequence repeats (SSRs) in the unigene sequences. Most SSRs were dinucleotides, among which AC/GT was the dominant SSR type. The molecular and genetic resources revealed in this study could be utilized for investigations on the fitness of the species in the marine environment and sustainability in a changing habitat.

Primary repair of untreated cleft palate in an elderly patient: a case report.

Untreated adult or elderly cleft lip and palate patients are rarely seen, but studies on delayed primary palatal closure have been performed in the less developed Asian and African countries, where access to medical care is difficult. A 64-year-old woman visited our clinic with untreated cleft palate with a 40×20-mm-wide defect in the medial palate. Two-flap palatoplasty under general anesthesia was performed to close the cleft palate. After 1 month, the result was favorable without any complications including oronasal fistula. Cleft palate primary repair in an elderly patient is rare and has some surgical problems that are associated with a wide range of defects, but good results can be obtained if surgery is performed well with appropriate considerations.

Molecular phylogenetic, population genetic and demographic studies of Nodularia douglasiae and Nodularia breviconcha based on CO1 and 16S rRNA.

Freshwater mussels belonging to the genus Nodularia (Family Unionidae) are known to be widely distributed in East Asia. Although phylogenetic and population genetic studies have been performed for these species, there still remain unresolved questions in their taxonomic status and biogeographic distribution pathways. Here, the nucleotide sequences of CO1 and 16S rRNA were newly determined from 86 N. douglasiae and 83 N. breviconcha individuals collected on the Korean Peninsula. Based on these data, we revealed the following results: (1) N. douglasiae can be divided into the three genetic clades of A (only found in Korean Peninsula), B (widely distributed in East Asia), and C (only found in the west of China and Russia), (2) the clade A is not an independent species but a concrete member of N. douglasiae given the lack of genetic differences between the clades A and B, and (3) N. breviconcha is not a subspecies of N. douglasiae but an independent species apart from N. douglasiae. In addition, we suggested the plausible scenarios of biogeographic distribution events and demographic history of Nodularia species.

Langerhans cell histiocytosis of the mandible: two case reports and literature review.

Langerhans cell histiocytosis (LCH) is a rare disorder characterized by the proliferation of dendritic cells resulting in local or systemic symptoms. The clinical symptoms of patients with Langerhans cell histiocytosis depend on the site and the degree of involvement. This article describes two case histories of unifocal bony Langerhans cell histiocytosis with mandibular involvement and further discusses the appropriate management of such via a review of the literature.

Biomechanical analysis of a changed posterior condylar offset under deep knee bend loading in cruciate-retaining total knee arthroplasty.

BACKGROUND: The conservation of the joint anatomy is an important factor in total knee arthroplasty (TKA). The restoration of the femoral posterior condylar offset (PCO) has been well known to influence the clinical outcome after TKA. OBJECTIVE: The purpose of this study was to determine the mechanism of PCO in finite element models with conservation of subject anatomy and different PCO of ±1, ±2, ±3 mm in posterior direction using posterior cruciate ligament-retaining TKA. METHODS: Using a computational simulation, we investigated the influence of the changes in PCO on the contact stress in the polyethylene (PE) insert and patellar button, on the forces on the collateral and posterior cruciate ligament, and on the quadriceps muscle and patellar tendon forces. The computational simulation loading condition was deep knee bend. RESULTS: The contact stresses on the PE insert increased, whereas those on the patellar button decreased as posterior condylar offset translated to the posterior direction. The forces exerted on the posterior cruciate ligament and collateral ligaments increased as PCO translated to the posterior direction. The translation of PCO in the anterior direction, in an equivalent flexion angle, required a greater quadriceps muscle force. CONCLUSIONS: Translations of the PCO in the posterior and anterior directions resulted in negative effects in the PE insert and ligament, and the quadriceps muscle force, respectively. Our findings suggest that orthopaedic surgeons should be careful with regard to the intraoperative conservation of PCO, because an excessive change in PCO may lead to quadriceps weakness and an increase in posterior cruciate ligament tension.

Transcriptome analysis of air-breathing land slug, Incilaria fruhstorferi reveals functional insights into growth, immunity, and reproduction.

BACKGROUND: Incilaria (= Meghimatium) fruhstorferi is an air-breathing land slug found in restricted habitats of Japan, Taiwan and selected provinces of South Korea (Jeju, Chuncheon, Busan, and Deokjeokdo). The species is on a decline due to depletion of forest cover, predation by natural enemies, and collection. To facilitate the conservation of the species, it is important to decide on a number of traits related to growth, immunity and reproduction addressing fitness advantage of the species. RESULTS: The visceral mass transcriptome of I. fruhstorferi was enabled using the Illumina HiSeq 4000 sequencing platform. According to BUSCO (Benchmarking Universal Single-Copy Orthologs) method, the transcriptome was considered complete with 91.8% of ortholog genes present (Single: 70.7%; Duplicated: 21.1%). A total of 96.79% of the raw read sequences were processed as clean reads. TransDecoder identified 197,271 contigs that contained candidate-coding regions. Of a total of 50,230 unigenes, 34,470 (68.62% of the total unigenes) annotated to homologous proteins in the Protostome database (PANM-DB). The GO term and KEGG pathway analysis indicated genes involved in metabolism, phosphatidylinositol signalling system, aminobenzoate degradation, and T-cell receptor signalling pathway. Many genes associated with molluscan innate immunity were categorized under pathogen recognition receptor, TLR signalling pathway, MyD88 dependent pathway, endogenous ligands, immune effectors, antimicrobial peptides, apoptosis, and adaptation-related. The reproduction-associated unigenes showed homology to protein fem-1, spermatogenesis-associated protein, sperm associated antigen, and testis expressed sequences, among others. In addition, we identified key growth-related genes categorized under somatotrophic axis, muscle growth, chitinases and collagens. A total of 4822 Simple Sequence Repeats (SSRs) were also identified from the unigene sequences of I. fruhstorferi. CONCLUSIONS: This is the first available genomic information for non-model land slug, I. fruhstorferi focusing on genes related to growth, immunity, and reproduction, with additional focus on microsatellites and repeating elements. The transcriptome provides access to greater number of traits of unknown relevance in the species that could be exploited for in-depth analyses of evolutionary plasticity and making informed choices during conservation planning. This would be appropriate for understanding the dynamics of the species on a priority basis considering the ecological, health, and social benefits.

Comparison of the biomechanical effect of posterior condylar offset and kinematics between posterior cruciate-retaining and posterior-stabilized total knee arthroplasty.

BACKGROUND: The effect of the changes in the femoral posterior condylar offset (PCO) on anterior-posterior (AP) translation and internal-external (IE) rotation in cruciate-retaining (CR) and posterior-stabilized (PS) total knee arthroplasty (TKA) remains unknown. The purpose of this study was to compare the kinematics in CR and PS TKA with respect to the difference in prosthetic design and PCO change through a computational simulation. METHODS: We developed three-dimensional finite element models with the different PCOs of ±1, ±2 and ±3 mm in the posterior direction using CR and PS TKA. We performed the simulation with different PCOs under a deep knee bend condition and evaluated the kinematics for the AP and IE in CR and PS TKA. RESULTS: The more tibiofemoral (TF) translation in the posterior direction was found as PCO translated in posterior direction for both CR and PS TKA compared to the neutral position. However, the change of the AP translation with respect to the PCO change in CR TKA was greater than PS TKA. The more TF external rotation was found as PCO translated in the anterior direction for both CR and PS TKA compared to the neutral position. However, unlike the TF translation, the TF rotation was not influenced by the PCO change in both CR and PS TKA. CONCLUSION: The PCO magnitude was influenced by a postoperative change in the kinematics in CR TKA although a relatively smaller effect was observed in PS TKA. Hence, surgeons should be aware of the PCO change, especially for CR TKA.

Medical adhesive related skin injury after dental surgery.

An 87-year-old woman was referred for the extraction of residual teeth and removal of tori prior to prosthetic treatment. After surgery under general anesthesia, the surgical tape was removed to detach the bispectral index sensor and the hair cover. After the surgical tape was removed, skin injury occurred on the left side of her face. After epidermis repositioning and ointment application, a dressing was placed over the injury. Her wound was found to have healed completely on follow-up examination. Medical adhesive related skin injury (MARSI) is a complication that can occur after surgery and subjects at the extremes of age with fragile skin are at a higher risk for such injuries. Careful assessment of the risk factors associated with MARSI is an absolute necessity.

Control of accumulated volatile fatty acids by recycling nitrified effluent.

BACKGROUND: Volatile fatty acids (VFA) often accumulate in anaerobic digestion systems, decreasing pH levels and causing unstable operational performance and poor biogas production. The aim of this study is to improve anaerobic digestion efficiency by controlling/reducing the accumulation of VFAs in a continuous anaerobic digestion system. METHODS: NO3 --N was added to the digester and its effects on VFAs were investigated. When the system reached an unstable condition with the accumulation of VFAs, the digester was fed at an organic loading rate of 6 kg COD (chemical oxygen demand)/m3∙d and 0.5Q of aeration tank effluent (1500 mg/L of NO3 --N) was recirculated. RESULTS: With the addition of NO3 --N, VFAs were utilized during denitrification, after which methane production started. Furthermore, the accumulated VFAs could be used as a carbon substrate by denitrifying bacteria. After 56 d, a normal VFA concentration could be achieved. Methane production was 0.02-0.03 L CH4/g VS higher with NO3 --N recirculation and feeding than that without feeding. CONCLUSIONS: The results show that the addition of NO3 --N is a potentially feasible method to control VFAs. Combined with recirculation and feeding, the method can be used to effectively prevent the inhibition of methanogenic microbial activities caused by accumulated VFAs and enhance denitrification and methane production in anaerobic digesters.

Effects of posterior condylar offset and posterior tibial slope on mobile-bearing total knee arthroplasty using computational simulation.

BACKGROUND: Postoperative changes of the femoral posterior condylar offset (PCO) and posterior tibial slope (PTS) affect the biomechanics of the knee joint after fixed-bearing total knee arthroplasty (TKA). However, the biomechanics of mobile-bearing is not well known. Therefore, the aim of this study was to investigate whether alterations to the PCO and PTS affect the biomechanics for mobile-bearing TKA. METHODS: We used a computational model for a knee joint that was validated using in vivo experiment data to evaluate the effects of the PCO and PTS on the tibiofemoral (TF) joint kinematics, patellofemoral (PF) contact stress, collateral ligament force and quadriceps force, for mobile-bearing TKA. The computational model was developed using ±1-, ±2- and ±3-mm PCO models in the posterior direction and -3°, 0°, +3°, and +6° PTS models based on each of the PCO models. RESULTS: The maximum PF contact stress, collateral ligament force and quadriceps force decreased as the PTS increased. In addition, the maximum PF contact stress and quadriceps force decreased, and the collateral ligament force increased as PCO translated in the posterior direction. This trend is consistent with that observed in any PCO and PTS. CONCLUSIONS: Our findings show the various effects of postoperative alterations in the PCO and PTS on the biomechanical results of mobile-bearing TKA. Based on the computational simulation, we suggest that orthopaedic surgeons intraoperatively conserve the patient's own anatomical PCO and PTS in mobile-bearing TKA.

Comparison of Kinematics in Cruciate Retaining and Posterior Stabilized for Fixed and Rotating Platform Mobile-Bearing Total Knee Arthroplasty with respect to Different Posterior Tibial Slope.

Reconstructed posterior tibial slope (PTS) plays a significant role in kinematics restoration after total knee arthroplasty (TKA). However, the effect of increased and decreased PTS on prosthetic type and design has not yet been investigated. We used a finite element model, validated using in vitro data, to evaluate the effect of PTS on knee kinematics in cruciate-retaining (CR) and posterior-stabilized (PS) fixed TKA and rotating platform mobile-bearing TKA. Anterior-posterior tibial translation and internal-external tibial rotation were investigated for PTS ranging from -3° to 15°, with increments of 1°, for three different designs of TKA. Tibial posterior translation and external rotation increased as the PTS increased in both CR and PS TKAs. In addition, there was no remarkable difference in external rotation between CR and PS TKAs. However, for the mobile-bearing TKA, PTS had less effect on the kinematics. Based on our computational simulation, PTS is the critical factor that influences kinematics in TKA, especially in the CR TKA. Therefore, the surgeon should be careful in choosing the PTS in CR TKAs.

Transcriptome analysis of the threatened snail Ellobium chinense reveals candidate genes for adaptation and identifies SSRs for conservation genetics.

Ellobium chinense (Pfeiffer, 1854) is a brackish pulmonate species that inhabits the bases of mangrove trees and is most commonly found in salt grass meadows. Threats to mangrove ecosystems due to habitat degradation and overexploitation have threatened the species with extinction. In South Korea, E. chinense has been assessed as vulnerable, but there are limited data on its population structure and distribution. The nucleotide and protein sequences for this species are not available in databases, which limits the understanding of adaptation-related traits. We sequenced an E. chinense cDNA library using the Illumina platform, and the subsequent bioinformatics analysis yielded 227,032 unigenes. Of these unigenes, 69,088 were annotated to matched protein and nucleotide sequences in databases, for an annotation rate of 30.42%. Among the predominant gene ontology terms, cellular and metabolic processes (under the biological process category), membrane and cell (under the cellular component category), and binding and catalytic activity (under the molecular function category) were noteworthy. In addition, 4850 unigenes were distributed to 15 Kyoto Encyclopaedia of Genes and Genomes based enrichment categories. Among the candidate genes related to adaptation, angiotensin I converting enzyme, adenylate cyclase activating polypeptide, and AMP-activated protein kinase were the most prominent. A total of 15,952 simple sequence repeats (SSRs) were identified in sequences of > 1 kb in length. The di- and trinucleotide repeat motifs were the most common. Among the repeat motif types, AG/CT, AC/GT, and AAC/GTT dominated. Our study provides the first comprehensive genomics dataset for E. chinense, which favors conservation programs for the restoration of the species and provides sufficient evidence for genetic variability among the wild populations.

The increase in posterior tibial slope provides a positive biomechanical effect in posterior-stabilized total knee arthroplasty.

PURPOSE: This study aims to clarify the influence of the posterior tibial slope (PTS) on knee joint biomechanics after posterior-stabilized (PS) total knee arthroplasty (TKA) using a computer simulation. METHODS: A validated TKA computational model was used to evaluate and quantify the effects of an increased PTS. In order to conduct a squat simulation, models with a - 3° to 15° PTS using increments of 3° were developed. Forces on the quadriceps and collateral ligament, a tibial posterior translation, contact point on a polyethylene (PE) insert, and contact stress on the patellofemoral (PF) joint and post in a PE insert were compared. RESULTS: The maximum force on the quadriceps and the PF contact stress decreased with increases in the PTS. The kinematics on the tibiofemoral (TF) joint translated in an increasingly posterior manner, and the medial and lateral contact points on a PE insert were located in posterior regions with increases in the PTS. Additionally, increases in the PTS decreased the force on the collateral ligament and increased the contact stress on the post in a PE insert. A higher force on the quadriceps is required when the PTS decreases with an equivalent flexion angle. CONCLUSIONS: A surgeon should be prudent in terms of determining the PTS because an excessive increase in the PTS may lead to the progressive loosening of the TF joint due to a reduction in collateral ligament tension and failure of the post in a PE insert. Thus, we support a more individualized approach of optimal PTS determination given the findings of the study.

Influence of Increased Posterior Tibial Slope in Total Knee Arthroplasty on Knee Joint Biomechanics: A Computational Simulation Study.

BACKGROUND: The reconstructed posterior tibial slope (PTS) plays a significant role in restoring knee kinematics in cruciate-retaining-total knee arthroplasty (TKA). A few studies have reported the effect of the PTS on biomechanics. METHODS: This study investigates the effect of the PTS on tibiofemoral (TF) kinematics, patellofemoral (PF) contact stress, and forces at the quadriceps, posterior cruciate ligament (PCL) and collateral ligament after cruciate-retaining-TKA using computer simulations. The simulation for the validated TKA finite element model was performed under deep knee bend condition. All analyses were repeated from -3° to 15° PTS in increments of 3°. RESULTS: The kinematics on the TF joint translated increasingly posteriorly when the PTS increased. Medial and lateral contact points translated in posterior direction in extension and flexion as PTS increased. The maximum contact stress on the PF joint and quadriceps, and collateral ligament force decreased when the PTS increased. An implantation of the tibial plate with increased PTS reduced the PCL load. Physiologic insert movement led to an increasingly posterior position of the femur and reduced quadriceps force especially for knee flexion angles above high flexion (120°) when compared to TKA with a decreased slope of the tibial base plate. CONCLUSION: An increase in the PTS increased medial and lateral movements without paradoxical motion. However, an excessive PTS indicated progressive loosening of the TF joint gap due to a reduction in collateral ligament tension during flexion.

Sequencing and de novo assembly of visceral mass transcriptome of the critically endangered land snail Satsuma myomphala: Annotation and SSR discovery.

Satsuma myomphala is critically endangered through loss of natural habitats, predation by natural enemies, and indiscriminate collection. It is a protected species in Korea but lacks genomic resources for an understanding of varied functional processes attributable to evolutionary success under natural habitats. For assessing the genetic information of S. myomphala, we performed for the first time, de novo transcriptome sequencing and functional annotation of expressed sequences using Illumina Next-Generation Sequencing (NGS) platform and bioinformatics analysis. We identified 103,774 unigenes of which 37,959, 12,890, and 17,699 were annotated in the PANM (Protostome DB), Unigene, and COG (Clusters of Orthologous Groups) databases, respectively. In addition, 14,451 unigenes were predicted under Gene Ontology functional categories, with 4581 assigned to a single category. Furthermore, 3369 sequences with 646 having Enzyme Commission (EC) numbers were mapped to 122 pathways in the Kyoto Encyclopedia of Genes and Genomes Pathway database. The prominent protein domains included the Zinc finger (C2H2-like), Reverse Transcriptase, Thioredoxin-like fold, and RNA recognition motif domain. Many unigenes with homology to immunity, defense, and reproduction-related genes were screened in the transcriptome. We also detected 3120 putative simple sequence repeats (SSRs) encompassing dinucleotide to hexanucleotide repeat motifs from >1kb unigene sequences. A list of PCR primers of SSR loci have been identified to study the genetic polymorphisms. The transcriptome data represents a valuable resource for further investigations on the species genome structure and biology. The unigenes information and microsatellites would provide an indispensable tool for conservation of the species in natural and adaptive environments.

Biomechanical Effects of Posterior Condylar Offset and Posterior Tibial Slope on Quadriceps Force and Joint Contact Forces in Posterior-Stabilized Total Knee Arthroplasty.

This study aimed to determine the biomechanical effect of the posterior condylar offset (PCO) and posterior tibial slope (PTS) in posterior-stabilized (PS) fixed-bearing total knee arthroplasty (TKA). We developed ±1, ±2, and ±3 mm PCO models in the posterior direction and -3°, 0°, 3°, and 6° PTS models using a previously validated FE model. The influence of changes in the PCO and PTS on the biomechanical effects under deep-knee-bend loading was investigated. The contact stress on the PE insert increased by 14% and decreased by 7% on average as the PCO increased and decreased, respectively, compared to the neutral position. In addition, the contact stress on post in PE insert increased by 18% on average as PTS increased from -3° to 6°. However, the contact stress on the patellar button decreased by 11% on average as PTS increased from -3° to 6° in all different PCO cases. The quadriceps force decreased by 14% as PTS increased from -3° to 6° in all PCO models. The same trend was found in patellar tendon force. Changes in PCO had adverse biomechanical effects whereas PTS increase had positive biomechanical effects. However, excessive PTS should be avoided to prevent knee instability and subsequent failure.

Transcriptome sequencing and de novo characterization of Korean endemic land snail, Koreanohadra kurodana for functional transcripts and SSR markers.

The Korean endemic land snail Koreanohadra kurodana (Gastropoda: Bradybaenidae) found in humid areas of broadleaf forests and shrubs have been considered vulnerable as the number of individuals are declining in recent years. The species is poorly characterized at the genomic level that limits the understanding of functions at the molecular and genetics level. In the present study, we performed de novo transcriptome sequencing to produce a comprehensive transcript dataset of visceral mass tissue of K. kurodana by the Illumina paired-end sequencing technology. Over 234 million quality reads were assembled to a total of 315,924 contigs and 191,071 unigenes, with an average and N50 length of 585.6 and 715 bp and 678 and 927 bp, respectively. Overall, 36.32 % of the unigenes found matches to known protein/nucleotide sequences in the public databases. The direction of the unigenes to functional categories was determined using COG, GO, KEGG, and InterProScan protein domain search. The GO analysis search resulted in 22,967 unigenes (12.02 %) being categorized into 40 functional groups. The KEGG annotation revealed that metabolism pathway genes were enriched. The most prominent protein motifs include the zinc finger, ribonuclease H, reverse transcriptase, and ankyrin repeat domains. The simple sequence repeats (SSRs) identified from >1 kb length of unigenes show a dominancy of dinucleotide repeat motifs followed with tri- and tetranucleotide motifs. A number of unigenes were putatively assessed to belong to adaptation and defense mechanisms including heat shock proteins 70, Toll-like receptor 4, AMP-activated protein kinase, aquaporin-2, etc. Our data provide a rich source for the identification and functional characterization of new genes and candidate polymorphic SSR markers in K. kurodana. The availability of transcriptome information ( http://bioinfo.sch.ac.kr/submission/ ) would promote the utilization of the resources for phylogenetics study and genetic diversity assessment.