Lateral Tissues Research Articles

Biomechanical impact of progressive meniscal extrusion on the knee joint: a finite element analysis

BackgroundWhile measuring meniscal extrusion quantitatively is an early risk factor for knee osteoarthritis (KOA), the biomechanics involved in this process are not well understood. This study aimed to investigate the effects of varying degrees of medial and lateral meniscal extrusion and their material softening on knee osteoarthritis progression.MethodsFinite element analysis (FEA) was utilized to simulate varying degrees of meniscal extrusion (1–5 mm) in 72 knee joint models, representing progressive meniscal degeneration and material softening due to injury. Changes in von Mises stress of the cartilage and menisci and the load distribution on the tibial plateau’s meniscus and cartilage were studied under balanced standing posture in both healthy and injured knees, and statistical analysis was performed using Spearman correlation.ResultsCompared to healthy knees, peak stress in medial compartment tissues increased by over 40% with 4 mm of medial meniscus extrusion, and in lateral compartment tissues with 2 mm of lateral meniscus extrusion. Meniscus extrusion reduced the contact load between the meniscus and femoral cartilage but increased it between the tibial and femoral cartilages, with a maximum increase up to fivefold. Spearman correlation analysis indicated that meniscal extrusion significantly affected peak stress and contact loads in the respective knee compartment (p < 0.001), with a lesser impact on the opposite compartment. Notably, medial meniscal extrusion also significantly increased peak stress in the lateral tibial cartilage (p < 0.05).ConclusionsThe quantitative analysis revealed that meniscal extrusion significantly affected the biomechanics of soft tissues within the same compartment, with limited impact on the opposite side. Specifically, Medial extrusion beyond 4 mm significantly affected the biomechanics of the medial compartment, while lateral extrusion over 2 mm had a similar impact on the lateral compartment. Meniscal softening, without altering joint contact characteristics, primarily affected the biomechanics of the meniscus itself, with minimal impact on other soft tissues.

The role of D3-type cyclins is related to cytokinin and the bHLH transcription factor SPATULA in Arabidopsis gynoecium development

Main conclusionWe studied the D3-type cyclin function during gynoecium development in Arabidopsis and how they are related to the hormone cytokinin and the transcription factor SPATULA.Growth throughout the life of plants is sustained by cell division and differentiation processes in meristematic tissues. In Arabidopsis, gynoecium development implies a multiphasic process where the tissues required for pollination, fertilization, and seed development form. The Carpel Margin Meristem (CMM) is a mass of undifferentiated cells that gives rise to the gynoecium internal tissues, such as septum, ovules, placenta, funiculus, transmitting tract, style, and stigma. Different genetic and hormonal factors, including cytokinin, control the CMM function. Cytokinin regulates the cell cycle transitions through the activation of cell cycle regulators as cyclin genes. D3-type cyclins are expressed in proliferative tissues, favoring the mitotic cell cycle over the endoreduplication. Though the role of cytokinin in CMM and gynoecium development is highly studied, its specific role in regulating the cell cycle in this tissue remains unclear. Additionally, despite extensive research on the relationship between CYCD3 genes and cytokinin, the regulatory mechanism that connects them remains elusive. Here, we found that D3-type cyclins are expressed in proliferative medial and lateral tissues. Conversely, the depletion of the three CYCD3 genes showed that they are not essential for gynoecium development. However, the addition of exogenous cytokinin showed that they could control the division/differentiation balance in gynoecium internal tissues and outgrowths. Finally, we found that SPATULA can be a mechanistic link between cytokinin and the D3-type cyclins. The data suggest that the role of D3-type cyclins in gynoecium development is related to the cytokinin response, and they might be activated by the transcription factor SPATULA.

The Tissue-Specific Eco-Exposome: Differential Pharmaceutical Bioaccumulation and Disposition in Fish among Trophic Positions.

Though bioaccumulation of pharmaceuticals by aquatic organisms continues to receive scientific attention, the internal disposition of these contaminants among different tissue compartments of fish species has been infrequently investigated, particularly among fish at different trophic positions. We tested a human to fish biological read-across hypothesis for contaminant disposition by examining tissue-specific accumulation in three understudied species, longnose gar (Lepisosteus osseus; piscivore), gizzard shad (Dorosoma cepedianum; planktivore/detritivore), and smallmouth buffalo (Ictiobus bubalus; benthivore), from a river influenced by municipal effluent discharge. In addition to surface water, fish plasma, and brain, gill, gonad, liver, and lateral muscle fillet tissues were analyzed via isotope dilution liquid chromatography tandem mass spectrometry. Caffeine and sucralose, two common effluent tracers, were quantitated at low micrograms per liter levels in surface water, while an anticonvulsant, carbamazepine, was observed at levels up to 37 ng/L. The selective serotonin reuptake inhibitors (SSRIs) fluoxetine and sertraline and primary metabolites were detected in at least one tissue of all three species at low micrograms per kilogram concentrations. Within each species, brain and liver of select fish contained the highest levels of SSRIs compared to plasma and other tissues, which is generally consistent with human tissue disposition patterns. However, we observed differential accumulation among specific tissue types and species. For example, mean levels of sertraline in brain and liver tissues were 13.4 µg/kg and 1.5 µg/kg in gizzard shad and 1.3 µg/kg and 7.3 µg/kg in longnose gar, respectively. In contrast, smallmouth buffalo did not consistently accumulate SSRIs to detectable levels. Tissue-specific eco-exposome efforts are necessary to understand mechanisms associated with such marked bioaccumulation and internal dispositional differences among freshwater fish species occupying different trophic positions. Environ Toxicol Chem 2024;43:1894-1902. © 2024 The Author(s). Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Seeding activity of human superoxide dismutase 1 aggregates in familial and sporadic amyotrophic lateral sclerosis postmortem neural tissues by real-time quaking-induced conversion

Amyotrophic lateral sclerosis (ALS) is a rapidly progressive neurodegenerative disease with average lifespan of 2–5 years after diagnosis. The identification of novel prognostic and pharmacodynamic biomarkers are needed to facilitate therapeutic development. Metalloprotein human superoxide dismutase 1 (SOD1) is known to accumulate and form aggregates in patient neural tissue with familial ALS linked to mutations in their SOD1 gene. Aggregates of SOD1 have also been detected in other forms of ALS, including the sporadic form and the most common familial form linked to abnormal hexanucleotide repeat expansions in the Chromosome 9 open reading frame 72 (C9ORF72) gene. Here, we report the development of a real-time quaking-induced conversion (RT-QuIC) seed amplification assay using a recombinant human SOD1 substrate to measure SOD1 seeding activity in postmortem spinal cord and motor cortex tissue from persons with different ALS etiologies. Our SOD1 RT-QuIC assay detected SOD1 seeds in motor cortex and spinal cord dilutions down to 10–5. Importantly, we detected SOD1 seeding activity in specimens from both sporadic and familial ALS cases, with the latter having mutations in either their SOD1 or C9ORF72 genes. Analyses of RT-QuIC parameters indicated similar lag phases in spinal cords of sporadic and familial ALS patients, but higher ThT fluorescence maxima by SOD1 familial ALS specimens and sporadic ALS thoracic cord specimens. For a subset of sporadic ALS patients, motor cortex and spinal cords were examined, with seeding activity in both anatomical regions. Our results suggest SOD1 seeds are in ALS patient neural tissues not linked to SOD1 mutation, suggesting that SOD1 seeding activity may be a promising biomarker, particularly in sporadic ALS cases for whom genetic testing is uninformative.

Optimizing Implant Width Selection in Breast Augmentation: Insights from On-Patient Landmark Positioning in 3D Breast Simulation.

The VECTRA® XT 3D simulation program is a valuable tool for implant selection during the decision-making process of breast augmentations. The software allows for the assignment of automatically or manually positioned anatomical landmarks and provides accompanying measurements. This study aimed to present a strategy for standardizing landmark placement on Vectra® images during primary breast augmentations, optimize implant selection, and insure consistency in breast width data collection. We performed a retrospective analysis of patient data wherein breast width measurements were obtained through Clinical Assessment (CA) and the Vectra® program using both automatically (VA) and manually (VM) placed landmarks. Consideration was also given to the thickness of medial and lateral breast tissues. The suggested implant base width from all groups was then compared to the actual implant (AI) used during surgical procedures. Comparing the change rates in relation to the actual implant width revealed that rates in CA measurements were significantly lower than those in VA (p < 0.05), and similarly lower than those in VM (p < 0.05). Furthermore, change rates in VM were significantly lower than in VA (p < 0.05). Each group showed a significant positive correlation with the actual implant width. The Vectra® XT 3D simulation program is a valuable adjunct in breast augmentation. However, relying on it solely may lead to inaccuracies in the assessment of breast dimensions due to automatic landmarking. To achieve standardized measurements, landmarks can be marked on the patient before creating the Vectra® image. Our results underline the superiority of clinical judgment over simulation programs in establishing a reliable surgical plan.

A high-resolution gene expression map of the medial and lateral domains of the gynoecium of Arabidopsis.

Angiosperms are characterized by the formation of flowers, and in their inner floral whorl, one or various gynoecia are produced. These female reproductive structures are responsible for fruit and seed production, thus ensuring the reproductive competence of angiosperms. In Arabidopsis (Arabidopsis thaliana), the gynoecium is composed of two fused carpels with different tissues that need to develop and differentiate to form a mature gynoecium and thus the reproductive competence of Arabidopsis. For these reasons, they have become the object of study for floral and fruit development. However, due to the complexity of the gynoecium, specific spatio-temporal tissue expression patterns are still scarce. In this study, we used precise laser-assisted microdissection and high-throughput RNA sequencing to describe the transcriptional profiles of the medial and lateral domain tissues of the Arabidopsis gynoecium. We provide evidence that the method used is reliable and that, in addition to corroborating gene expression patterns of previously reported regulators of these tissues, we found genes whose expression dynamics point to being involved in cytokinin and auxin homeostasis and in cell cycle progression. Furthermore, based on differential gene expression analyses, we functionally characterized several genes and found that they are involved in gynoecium development. This resource is available via the Arabidopsis eFP browser and will serve the community in future studies on developmental and reproductive biology.

Strong, tough, anisotropic, flexible, and transparent bamboo films

Bamboo has higher strength, better toughness, and a shorter growth cycle than wood. It is of great significance to use top-down nanotechnology to construct high-performance bamboo film (BF) instead of petroleum-based plastic film. However, bamboo lacks lateral cell tissues, and its structure is prone to disintegration when the components are removed. Herein, slice bamboo veneers (SBV) were used as building blocks, and a sequential process of alkali pretreatment-crosslinking-delignification-compression densification was conducted to fabricate the high-performance BF. The oriented arrangement of nanofibers, highly dense microstructure, and a large number of hydrogen bonds between cellulose microfibrils endowed BF with anisotropic and excellent mechanical and optical properties. The longitudinal tensile strength and toughness of the prepared BF were 656.6 MPa and 33.7 MJ m−3, respectively, and its light transmittance and haze were 70.2% and 88.1%, respectively. In addition, BF has good thermal insulation with a thermal conductivity of 0.0857 W m−1 K−1. The combined characteristics of BF, including high strength, high toughness, flexibility, anisotropy, high optical transmittance, and thermal insulation, make them suitable to be applied as optoelectronic substrates, insulating building materials, and green packaging materials.

First report of 'Candidatus Phytoplasma asteris' associated with witches'-broom and plexus bud disease of Cerasus serrula in China.

Cherry blossoms (Cerasus serrula) are native to the temperate zone around the Himalayas in the northern hemisphere, mainly distributed in the west and southwest of China, including Yunnan, Sichuan and Tibet. Cherry has high ornamental, edible and medicinal value. In August 2022, we observed that Cherry trees exhibited witches' broom and plexus bud in Kunming City, Yunan Province, China. The symptoms consisted of many small branches with little leaves at the top of branches, stipule lobation, and clustered adventitious buds that are tumor-like on the branches that usually cannot sprout normally. As disease intensity increased, the branches dried up from the top to the bottom till the death of the whole plant. We named this disease C. serrula witches' broom disease (CsWB). We found CsWB in the areas of Panlong, Guandu, Xishan Districts in Kunming, where more than 17% of the plants we surveyed were infected. We collected 60 samples from across the three districts. These included 15 symptomatic and 5 asymptomatic plants per district. The lateral stem tissues were observed under a scanning electron microscope (Hitachi S-3000N). The nearly spherical bodies were found in the phloem cells of symptomatic plants. Total DNA extraction was conducted from 0.1 g tissue using the CTAB method (Porebski et al. 1997), ddH2O was used as the negative control, and Dodonaea viscose plants with witches' broom symptoms were used as the positive control. The nested PCR was used to amplify the 16S rRNA gene (Lee et al. 1993; Schneider et al. 1993) and PCR amplicon of 1.2 kb were amplified (GenBank accessions: OQ408098; OQ408099; OQ408100). The direct PCR specific to the ribosomal protein (rp) gene yielded amplicons of approximately 1.2 kb with primer pair rp(I)F1A and rp(I)R1A (Lee et a. 2003) (GenBank accessions: OQ410969; OQ410970; OQ410971). The fragment from 33 symptomatic samples was consistent with the positive control, and absent for asymptomatic samples, suggesting an association of phytoplasma with the disease. A BLAST analysis of the 16S rRNA sequences of CsWB phytoplasma showed that it has a 99.76% similarity with Trema laevigata witches' broom phytoplasma (GenBank accession MG755412). The rp sequence shared 99.75% identity with Cinnamomum camphora witches' broom phytoplasma (GenBank accession OP649594). An analysis with iPhyClassifier showed that the virtual RFLP pattern derived from the 16S rDNA sequence shares 99.3% similarity with that of the 'Ca. Phytoplasma asteris' reference strain (GenBank accession: M30790), and the virtual RFLP pattern derived from the fragment is identical (similarity coefficient 1.00) to the reference pattern of 16Sr group I, subgroup B (GenBank accession: AP006628). Thus, CsWB phytoplasma is identified as 'Ca. Phytoplasma asteris'-related strain belonging to sub-group 16SrI-B. The phylogenetic tree was constructed based on 16S rRNA gene and rp gene sequences by using MEGA version 6.0 (Tamura et al. 2013) with neighbor-joining (NJ) method and bootstrap support was estimated with 1000 replicates. The result indicated that the CsWB phytoplasma formed a subclade in 16SrI-B and rpI-B respectively. In addition, the clean 1-year-old C. serrula were tested positive for the phytoplasma using the nested PCR 30 days after being grafted with naturally infected twigs with CsWB symptoms. To the best of our knowledge, Cherry blossoms is a new host of 'Ca. Phytoplasma asteris'-related strains in China. The newly emerged disease is a threat to the ornamental value of cherry blossoms and the production of wood quality.

Features of regional relapses in the central zone after surgical treatment of highly differentiated thyroid cancer

Introduction. Thyroid cancer is the most common malignant tumor of the endocrine system and comprises 33 % of malignant neoplasms of the head and neck. first echelon lymph nodes affected by this pathology are located in the central area of the neck. The metastatic process further advances into the lymph nodes in the lateral tissues of the neck. Currently, there is no consensus on the effectiveness of prophylactic central lymph node dissection and no conclusive criteria determining the risk of recurrence in the central zone.Aim. The assessment of factors influencing the risk of regional recurrence in the central zone.Materials and methods. The study included 30 patients with highly differentiated thyroid cancer, in whom metastatic nodes in the central zone were identified and verified during primary treatment. All patients previously underwent unilateral or 2-sided central lymph node dissection in various institutions. During the period of dynamic observation, these patients revealed regional recurrence in the central zone. patients were observed and treated for recurrence at the National medical Research Center of Oncology named after N.N. Blokhin of the ministry of Health of Russia.Results. primary surgical treatment in a specialized oncological institution was performed in 21 (70 %) patients, in a general surgical medical institution – in 7 (23.3 %), in an endocrinological institution – in 2 (6.7 %) patients. when assessing the influence of the morphological type and variant of thyroid cancer on the risk of developing regional recurrence, no statistical significance was found. No statistically significant factors were found in the analysis of the influence of characteristics of the primary tumor, such as tumor size, multifocal lesion, vascular invasion, extrathyroidal spread, on the risk of regional recurrence. In 16 (53.3 %) patients, recurrence occurred in the paratracheal zone of the ipsilateral side, in 7 (23.3 %) patients in the paratracheal zone of the contralateral side, bilateral lesions were observed in 2 (6.67 %) cases. 3 (10 %) patients were diagnosed with transient parathyroid insufficiency, 1 (3.3 %) – permanent parathyroid insufficiency, 1 (3.3 %) – hematoma in the area of the postoperative wound, patient, 2 (6.6 %) – injury to the recurrent laryngeal nerve.Conclusion. Residual metastatic lymph nodes after previous surgical treatment are the reason for performing a second operation, which carries both the risks of postoperative complications and the psychological burden on the patient. Thus, an adequate and radically performed primary surgical intervention is the key to reducing regional recurrences.

First report of 'Candidatus Phytoplasma asteris' associated with witches'-broom disease of Pinus yunnanensis in China.

Pinus yunnanensis is an evergreen tree belonging to Pinaceae. The species is distributed in the east of Tibet, southwest of Sichuan, southwest of Yunnan, southwest of Guizhou and northwest of Guangxi. It is an indigenous and pioneer tree species for barren mountain afforestation in southwest China. P. yunnanensis has important value to both the building and medicine industries (Liu et al. 2022). In May 2022, P. yunnanensis showing witches'-broom symptom were found in Panzhihua City, Sichuan Province, China. The symptomatic plants had yellow or red needle, and exhibited plexus bud and needle wither. The lateral buds of infected pines developed into twigs. Some lateral buds grew in clusters and a few sprouted needles (Fig.1). The disease was named the P. yunnanensis witches'-broom disease (PYWB) and was found in some areas of Miyi, Renhe, and Dongqu. More than 9% of the pines showed these symptoms in the three areas surveyed, and the disease was spreading. A total of 39 samples were collected from three areas, including 25 symptomatic plants and 14 asymptomatic plants, respectively. The lateral stem tissues of 18 samples were observed under a scanning electron microscope (Hitachi S-3000N). Spherical bodies were found in the phloem sieve cells of symptomatic pines (Fig.1). Total DNA was extracted from 18 plant samples using the CTAB method (Porebski et al. 1997) and subjected to nested-PCR testing. Double-distilled water and DNA extracted from asymptomatic plants were used as negative controls, and DNA extracted from Dodonaea viscosa affected by the D. viscosa witches'-broom disease was used as positive control. Nested PCR was employed to amplify the pathogen's 16S rRNA gene (Lee et al. 1993; Schneider et al. 1993) and 1.2 kb segment were produced (GenBank accessions OP646619; OP646620; OP646621). PCR specific to the ribosomal protein (rp) gene yielded segment of approximately 1.2 kb (Lee et al. 2003)(GenBank accessions OP649589; OP649590; OP649591). The fragment size from 15 samples was consistent with the positive control, confirming the association of phytoplasma with the disease. A BLAST analysis of the 16S rRNA sequences of P. yunnanensis witches'-broom phytoplasma showed that it shared 99.12% ~99.76% identity with that of Trema laevigata witches'-broom phytoplasma (GenBank accession MG755412). The rp sequence shared 99.84%~99.92% identity with that of Cinnamomum camphora witches'-broom phytoplasma (GenBank accession OP649594). An analysis with iPhyClassifier (Zhao et. 2013) showed that the virtual RFLP pattern derived from OP646621 16S rDNA fragment of PYWB phytoplasma is identical (similarity coefficient 1.00) to the reference pattern of 16Sr group I, subgroup B (OY-M, GenBank accession AP006628). The phytoplasma is identified as a 'Candidatus Phytoplasma asteris'-related strain belonging to sub-group 16SrI-B. Interestingly, compared to AP006628, the virtual RFLP pattern derived from OP646619 and OP646620 fragments exhibit differences in three and one cleavage site, with a similarity coefficient of 0.92 and 0.97, respectively (Fig.2). These strains may represent a new subgroup within the 16Sr group I. The phylogenetic tree was reconstructed based on 16S rRNA and rp gene sequences using MEGA versio6.0 (Tamura et al. 2013). The analysis was conducted using the neighbor-joining (NJ) method with 1,000 replicates of bootstrap analysis. The results indicated that the PYWB phytoplasmas grouped into clades including phytoplasmas belonging to 16SrI-B and rpI-B, respectively (Fig.3). In addition, 2-year-old P. yunnanensis were used for grafting assays in nursery, and the twigs from infected pine under natural conditions were used as a scion, and the phytoplasma were detected using nested PCR after grafting for 40 d (Fig.4). In 2008-2014, P. sylvestris and P. mugo in Lituania had excessive branching symptoms that were attributed to 'Ca. Phtyoplasma Pini' (16SrXXI-A) or asteris' (16SrI-A) strains (Valiunas et al. 2015). In 2015, P. pungens with abnormal shoot branching in Maryland were found to be infected by 'Ca. Phytoplasma pini' strain (16SrXXI-B) (Costanzo et al. 2016). To the best of our knowledge, P. yunnanensis is a new host of 'Ca. Phytoplasma asteris'-related strain (16SrI-B) in China. The newly emerged disease is a threat to pines.

Chemiluminescence-Based Assay to Monitor Early Oxidative Bursts in Soybean (Glycine max) Lateral Roots.

The reactive oxygen species (ROS) burst assay is a valuable tool for studying pattern-triggered immunity (PTI) in plants. During PTI, the interaction between pathogen recognition receptors (PRRs) and pathogen-associated molecular patterns (PAMPs) leads to the rapid production of ROS in the apoplastic space. The resultant ROS can be measured using a chemiluminescent approach that involves the usage of horseradish peroxidase and luminol. Although several methods and protocols are available to detect early ROS bursts in leaf tissues, no dedicated method is available for root tissues. Here, we have established a reliable method to measure the PAMP-triggered ROS burst response in soybean lateral roots. In plants, lateral roots are the potential entry and colonization sites for pathogens in the rhizosphere. We have used important PAMPs such as chitohexaose, flagellin 22 peptide fragment, and laminarin to validate our method. In addition, we provide a detailed methodology for the isolation and application of fungal cell wall components to monitor the oxidative burst in soybean lateral roots. Furthermore, we provide methodology for performing ROS burst assays in soybean leaf discs with laminarin and fungal cell walls. This approach could also be applied to leaf and root tissues of other plant species to study the PTI response upon elicitor treatment. © 2023 Wiley Periodicals LLC. Basic Protocol: Reactive oxygen species (ROS) burst assay in soybean lateral root tissues Alternate Protocol: ROS burst assay in soybean leaf discs Support Protocol: Isolating fungal cell wall fractions.

Comparative transcriptome analysis on wild-simulated ginseng of different age revealed possible mechanism of ginsenoside accumulation

Panax ginseng is one of the most famous pharmaceutical plants in Asia. Ginseng plants grown in mountain have longer longevity which ensures higher accumulation of ginsenoside components than those grown in farms. However, wild-simulated ginseng over certain age cannot be easily distinguished in morphology. To identify transcriptomic mechanism of ginsenoside accumulation in older wild-simulated ginseng without large phenotype change, we performed comparative transcriptome analysis for leaf, shoot, and root tissues of 7-yr-old and 13yr-old wild-simulated ginseng. Of 559 differentially expressed genes (DEGs) in comparison between 7-yr-old and 13yr-old wild-simulated ginseng, 280 leaf-, 103 shoot-, and 164 root-mainly expressing genes were found to be changed in transcript level according to age. Functional analysis revealed that pentose-phosphate shunt and abscisic acid responsive genes were up-regulated in leaf tissues of 7-yr-old ginseng while defense responsive genes were up-regulated in root tissues of 13-yr-old ginseng. Quantitative real-time PCR revealed that jasmonic acid responsive genes, ERDL6, and some UGTs were up-regulated in 13-yr-old ginseng in higher order lateral root tissues. These data suggest that bacterial stimulation in mountain region can enhance the expression of several genes which might support minor ginsenoside biosynthesis.

First report of 'Candidatus Phytoplasma asteris' associated with witches'-broom disease of Cinnamomum camphora in China.

Camphor tree (Cinnamomum camphora) has a wide distribution in the world and is mainly distributed in the South and southwest in China. It can be used as both a wood and a medicine, with high value in industry, medicine, and ecology. In May 2022, it was observed that the approximately ten to fifteen years old Camphor trees were exhibiting witches'-broom, small leaf morphology and chlorosis, and leaf drop in Panzhihua City, Sichuan Province, China. The witches'-broom symptoms consisted of many small branches with little leaves at the top of branches. It was named C. camphora witches'-broom disease (CCWB) and was found in some areas of Miyi, Yanbian, Renhe Xiqu and Dongqu counties. More than 28% of the plants were infected on the five areas surveyed. Total 100 samples were collected from five areas, with 15 symptomatic plants and 5 asymptomatic plants each area. The lateral stem tissues were observed under a scanning electron microscope (Hitachi S-3000N). The nearly spherical bodies were found in the phloem sieve cells of symptomatic plants. Total DNA extraction was conducted from 0.1 g tissue using the CTAB method (Porebski et al. 1997), ddH2O was used as the negative control, and Dodonaea viscose witches'-broom disease plants were used as the positive control. The nested PCR was employed to amplify the 16S rRNA gene (Lee et al. 1993; Schneider et al. 1993) and PCR amplicon of 1.2 kb were amplified (GenBank accessions OP662614; OP662615; OP662616). The direct PCR specific to the ribosomal protein (rp) gene yielded amplicons of approximately 1.2 kb with primer pair rp(I)F1A and rp(I)R1A (Lee et al. 2003) (GenBank accessions OP649592; OP649593; OP649594). The fragment from 25 symptomatic samples was consistent with the positive control, and asymptomatic plants were negative, confirming an association of a phytoplasma with the disease. A BLAST analysis of the 16S rRNA sequences of CCWB phytoplasma showed that it has a 99.44% similarity with Trema laevigata witches'-broom phytoplasma (GenBank accession MG755412). The rp sequence shared 99.59% identity with rapeseed phyllody phytoplasma (GenBank accession CP055264). An analysis with iPhyClassifier showed that the virtual RFLP pattern derived from the query 16S rDNA fragment of CCWB phytoplasma is most similar to the reference pattern of the 16Sr group I, subgroup B (OY-M, GenBank accession AP006628). The phytoplasma is identified as 'Ca. Phytoplasma asteris'-related strain belonging to sub-group 16SrI-B. The phylogenetic tree was constructed based on 16S rRNA gene and rp gene sequences by using MEGA version 6.0 (Tamura et al. 2013) with neighbor-joining (NJ) method and bootstrap support was estimated with 1000 replicates. The result indicated that the CCWB phytoplasma formed a subclade in 16SrI-B and rpI-B respectively. In addition, the plants were positive for the phytoplasma using nested PCR after grafting for 30 d in nursery conditions. It is noteworthy that the plants were seriously damaged by aphid, Psyllidae and Ceroplastes. It is speculated that the insects of Homoptera typically transmit phytoplasmas by feeding on plant sap, thus it is necessary to control aphids in order to control the C. camphora witches'-broom disease. To the best of our knowledge, Camphor tree is a new host plant of 'Ca. Phytoplasma asteris' in China. The newly emerged disease is a threat to Camphor tree production.

Scarless Donor Site for Breast Reconstruction by Endoscopically Assisted Extended Latissimus Dorsi Flap plus Lipofilling.

Breast reconstruction using endoscopically assisted latissimus dorsi (LD) flap leaves no scar on the back; however, the small amount of tissue obtained makes this procedure less practical. This study aimed to propose a new technique of endoscopically assisted extended LD (eeLD) flap plus lipofilling, which could secure a large breast volume. Lateral thoracic adipose tissues supplied by the thoracodorsal artery branches and the LD muscle were elevated as a single unit only through the mastectomy scar and three ports through the lateral chest. Furthermore, fat was simultaneously injected to support the volume and shape of the breast. Changes in the volume of the reconstructed breast over time were measured using three-dimensional stereophotogrammetry. Overall, 15 breasts of 14 patients who underwent breast reconstruction using an eeLD flap exhibited no serious complications. On average, 281.9 ± 32.4 g of flap and 74.7 ± 19.4 mL of lipofilling were used. Within 8 weeks after the procedure, the volume of the reconstructed breast decreased to 69.5 ± 7.5% and then plateaued. Seven patients needed a subsequent session of lipofilling to acquire adequate breast volume and projection. Notably, according to the BREAST-Q back scores, patients who underwent eeLD flap surgery were significantly more satisfied than those who underwent conventional LD musculocutaneous flap surgery using a skin paddle on the back at the same institution (82.8 ± 9.2 versus 62.6 ± 6.3; P < 0.0001). Despite the limitations in volume, eeLD flap plus lipofilling is advantageous because it does not leave a noticeable donor-site scar. Therapeutic, IV.

PTEN inhibitor bisperoxovanadium protects against noise-induced hearing loss.

Studies have shown that phosphatase and tensin homolog deleted on chromosome ten (PTEN) participates in the regulation of cochlear hair cell survival. Bisperoxovanadium protects against neurodegeneration by inhibiting PTEN expression. However, whether bisperoxovanadium can protect against noise-induced hearing loss and the underlying mechanism remains unclear. In this study, we established a mouse model of noise-induced hearing loss by exposure to 105 dB sound for 2 hours. We found that PTEN expression was increased in the organ of Corti, including outer hair cells, inner hair cells, and lateral wall tissues. Intraperitoneal administration of bisperoxovanadium decreased the auditory threshold and the loss of cochlear hair cells and inner hair cell ribbons. In addition, noise exposure decreased p-PI3K and p-Akt levels. Bisperoxovanadium preconditioning or PTEN knockdown upregulated the activity of PI3K-Akt. Bisperoxovanadium also prevented H2O2-induced hair cell death by reducing mitochondrial reactive oxygen species generation in cochlear explants. These findings suggest that bisperoxovanadium reduces noise-induced hearing injury and reduces cochlear hair cell loss.

Scalable, large-size, and flexible transparent bamboo

Transparent wood (TW) has received extensive attention recently because of its excellent optical, mechanical, and thermal properties. However, the long growth cycle of timber and forbidden deforestation regulation limit the large-scale application. Bamboo has a much shorter growth cycle than wood, but its high density and lack of lateral cell tissues make it challenging to produce transparent products. Meanwhile, fabrication of large-size transparent bamboo (TB) remains a huge challenge because of the inherent features of irregular, hollow cylindrical shape with nodes, and big culm taper. In this work, the “alkali pretreatment-crosslinking-delignification” strategy was developed to fabricate the scalable, large-size, and flexible TB from the building block of sliced bamboo veneer (SBV). The optical transmittance and haze of the TB with a thickness of 1.0 mm were approximately 80 % and 72 %, respectively, and possessed the light modulation capability. The fabricated TB exhibited flexibility, and the tensile strength was 78.5 MPa, which was higher than all reported flexible TW. Besides, the TB exhibited a low thermal conductivity of 0.35 W m−1 k−1, which had an outstanding thermal insulation and indoor temperature regulation performance. More importantly, this study demonstrated the feasibility of successful preparation of large-size TB, which can promote its large-scale and industrial application. The TB with excellent overall performance holds great potential in engineering applications, especially for energy-efficient windows, light-tunable devices, flexible transparent materials, etc.

Poster 231: Lateral Extra-articular Tenodesis Provides Supraphysiological Restraint to Internal Tibial Rotation: In vitro Biomechanical Assessment

Objectives:Lateral extra-articular tenodesis (LET) reduces ACL graft failure rates two years after surgery when performed as an adjunct to ACL reconstruction (ACLR). Interestingly, previous biomechanical studies have shown that LET may reduce tibial rotation beyond that of the intact knee, while others found no such kinematic overconstraint. Parameters of ligament engagement have proven useful in characterizing the biomechanical function of the ACL and the anterolateral ligament; however, they have not been used to describe the biomechanics of LET. In this study, we compared engagement parameters (engagement point, in situ stiffness, and tissue force at peak applied load) of an LET-reconstructed knee compared to the native lateral tissues in response to an internal rotation torque at 0°, 30°, 60°, and 90° of knee flexion.Methods:Seven cadaveric knees (mean age: 39 ± 12; range: 28-54; 4 male) were mounted to a robotic manipulator. The robot applied an internal rotation torque of 5 Nm while monitoring the resulting internal tibial rotation (ITR) (in degrees). Each knee was tested following a bone-patellar tendon-bone ACL reconstruction with intact lateral tissues (consisting of the anterolateral ligament and Kaplan fibers) and after sectioning these tissues and performing LET (modified Lemaire technique). Resultant forces carried by the native lateral tissues and the LET were determined via superposition. The parameters of engagement were determined for both the native lateral tissues and the LET and compared via two-way repeated measures ANOVA (p < 0.05).Results:During an internal rotation test at full extension (0° of flexion), both the LET-reconstructed and native lateral tissues did not engage. At 30°, 60°, and 90° knee flexion, the native lateral tissues exhibited more in situ slack than the LET-reconstructed lateral tissues. Specifically, the native lateral tissues had 8° (p < 0.001), 13° (p < 0.001), and 14° (p < 0.001) more in situ slack than the LET-reconstructed lateral tissues at 30°, 60°, and 90° knee flexion, respectively. At 30° of flexion, the LET-reconstructed lateral tissues were 9° (p < 0.001) and 10° (p < 0.001) more slack than at 60° and 90° knee flexion. Across all three tested knee flexion angles (30°, 60°, and 90°), the LET-reconstructed lateral tissues had greater in situ stiffness than the native lateral tissues. The LET carried greater force at the peak applied internal rotation torque by 29 N at 30° of flexion (p = 0.006), but no statistical differences were identified at the other flexion angles.Conclusions:LET creates a supraphysiologic restraint to the native lateral tissues by engaging with less internal tibial rotation than the native lateral tissue at all flexion angles tested but full extension. The LET also carried greater force at the peak applied load and had a greater in situ stiffness at 30° of flexion than the native lateral tissues. On the whole, LET is a supraphysiological restraint to internal tibial rotation at 30° of flexion. The engagement point of LET may be modified surgically by altering the flexion angle, degree of tibial rotation at which the tenodesis is fixed, and/or the tension applied. Thus, discrepancies between previous biomechanical studies may arise from variations in one or more of these modifiable surgical parameters.Figure 1.Mean tissue force versus internal tibial rotation response of the LET and native lateral tissues with applied internal rotation torque at 0,30,60, and 90° of knee flexion. Whiskers represent one standard deviation. Engagement point denoted in yellow. Tissue force at peak applied internal tibial rotation torque denoted in blue.

Evidence for a Role of the Lateral Ectoderm in Drosophila Mesoderm Invagination.

The folding of two-dimensional epithelial sheets into specific three-dimensional structures is a fundamental tissue construction mechanism in animal development. A common mechanism that mediates epithelial folding is apical constriction, the active shrinking of cell apices driven by actomyosin contractions. It remains unclear whether cells outside of the constriction domain also contribute to folding. During Drosophila mesoderm invagination, ventrally localized mesoderm epithelium undergoes apical constriction and subsequently folds into a furrow. While the critical role of apical constriction in ventral furrow formation has been well demonstrated, it remains unclear whether, and if so, how the laterally localized ectodermal tissue adjacent to the mesoderm contributes to furrow invagination. In this study, we combine experimental and computational approaches to test the potential function of the ectoderm in mesoderm invagination. Through laser-mediated, targeted disruption of cell formation prior to gastrulation, we found that the presence of intact lateral ectoderm is important for the effective transition between apical constriction and furrow invagination in the mesoderm. In addition, using a laser-ablation approach widely used for probing tissue tension, we found that the lateral ectodermal tissues exhibit signatures of tissue compression when ablation was performed shortly before the onset of mesoderm invagination. These observations led to the hypothesis that in-plane compression from the surrounding ectoderm facilitates mesoderm invagination by triggering buckling of the mesoderm epithelium. In support of this notion, we show that the dynamics of tissue flow during mesoderm invagination displays characteristic of elastic buckling, and this tissue dynamics can be recapitulated by combining local apical constriction and global compression in a simulated elastic monolayer. We propose that Drosophila mesoderm invagination is achieved through epithelial buckling jointly mediated by apical constriction in the mesoderm and compression from the neighboring ectoderm.

Missense mutation of a class B heat shock factor is responsible for the tomato bushy root-2 phenotype

The bushy root-2 (brt-2) tomato mutant has twisting roots, and slower plant development. Here we used whole genome resequencing and genetic mapping to show that brt-2 is caused by a serine to cysteine (S75C) substitution in the DNA binding domain (DBD) of a heat shock factor class B (HsfB) encoded by SolycHsfB4a. This gene is orthologous to the Arabidopsis SCHIZORIZA gene, also known as AtHsfB4. The brt-2 phenotype is very similar to Arabidopsis lines in which the function of AtHsfB4 is altered: a proliferation of lateral root cap and root meristematic tissues, and a tendency for lateral root cap cells to easily separate. The brt-2 S75C mutation is unusual because all other reported amino acid substitutions in the highly conserved DBD of eukaryotic heat shock factors are dominant negative mutations, but brt-2 is recessive. We further show through reciprocal grafting that brt-2 exerts its effects predominantly through the root genotype even through BRT-2 is expressed at similar levels in both root and shoot meristems. Since AtHsfB4 is induced by root knot nematodes (RKN), and loss-of-function mutants of this gene are resistant to RKNs, BRT-2 could be a target gene for RKN resistance, an important trait in tomato rootstock breeding.Gene & accession numbersSolycHsfB4a - Solyc04g078770.

MTHFR promoter methylation might mitigate the effect of smoking at the level of LINE-1 in cleft lip tissues: A preliminary study.

The medial and maxillary aspects of the upper lip originate at separate embryonic stages and therefore may experience different maternal exposure patterns which may affect methylation. Based on this hypothesis, we investigated the level of methylation of the methylene tetrahydrofolate reductase promoter gene (mMTHFR) in tissues from cleft lip, and mMTHFR levels by MTHFR c.677C > T genotype. We further investigated whether mMTHFR mitigates the effect of smoking on long interspersed nuclear element (LINE-1) methylation in these tissues. DNA extracted from medial and lateral tissues of 26 infants with nonsyndromic cleft lip with or without cleft palate (nsCL/P) was bisulfite converted and mMTHFR was measured on a pyrosequenser. LINE-1 methylation and MTHFR c.677C > T genotype data were obtained in our previous study. There was no substantial difference in mMTHFR (p=.733) and LINE-1 (p=.148) between the two tissues. mMTHFR was not influenced by MTHFR c.677C > T genotype, but there was suggestive evidence that the difference was larger among infants exposed to maternal smoking compared to nonexposed. LINE-1 methylation differences were significant (p=.025) in infants born to nonsmoking mothers, but this was not apparent (p=.872) in infants born to mothers who smoked. Our Pearson's correlation analysis suggested a weak inverse association between mMTHFR and LINE-1 (r=-.179, p=.381). Our preliminary observation of differences in patterns of mMTHFR levels in lip tissue suggests the interplay of gene and environment in the establishment of methylation in tissues at both sides of cleft lip. This requires investigation in a larger cohort, integrated with metabolic assessment.