Elucidating the callus-to-shoot-forming mechanism in Capsicum annuum 'Dempsey' through comparative transcriptome analyses.

BACKGROUND: The formation of shoots plays a pivotal role in plant organogenesis and productivity. Despite its significance, the underlying molecular mechanism of de novo regeneration has not been extensively elucidated in Capsicum annuum 'Dempsey', a bell pepper cultivar. To address this, we performed a comparative transcriptome analysis focusing on the differential expression in C. annuum 'Dempsey' shoot, callus, and leaf tissue. We further investigated phytohormone-related biological processes and their interacting genes in the C. annuum 'Dempsey' transcriptome based on comparative transcriptomic analysis across five species. RESULTS: We provided a comprehensive view of the gene networks regulating shoot formation on the callus, revealing a strong involvement of hypoxia responses and oxidative stress. Our comparative transcriptome analysis revealed a significant conservation in the increase of gene expression patterns related to auxin and defense mechanisms in both callus and shoot tissues. Consequently, hypoxia response and defense mechanism emerged as critical regulators in callus and shoot formation in C. annuum 'Dempsey'. Current transcriptome data also indicated a substantial decline in gene expression linked to photosynthesis within regenerative tissues, implying a deactivation of the regulatory system governing photosynthesis in C. annuum 'Dempsey'. CONCLUSION: Coupled with defense mechanisms, we thus considered spatial redistribution of auxin to play a critical role in the shoot morphogenesis via primordia outgrowth. Our findings shed light on shoot formation mechanisms in C. annuum 'Dempsey' explants, important information for regeneration programs, and have broader implications for precise molecular breeding in recalcitrant crops.

Agrobacterium-Mediated Capsicum annuum Gene Editing in Two Cultivars, Hot Pepper CM334 and Bell Pepper Dempsey.

Peppers (Capsicum annuum L.) are the most widespread and cultivated species of Solanaceae in subtropical and temperate countries. These vegetables are economically attractive worldwide. Although whole-genome sequences of peppers and genome-editing tools are currently available, the precision editing of peppers is still in its infancy because of the lack of a stable pepper transformation method. Here, we employed three Agrobacterium tumefaciens strains-AGL1, EHA101, and GV3101-to investigate which Agrobacterium strain could be used for pepper transformation. Hot pepper CM334 and bell pepper Dempsey were chosen in this study. Agrobacterium tumefaciens GV3101 induced the highest number of calli in cv. Dempsey. All three strains generated similar numbers of calli for cv. CM334. We optimized a suitable concentration of phosphinothricin (PPT) to select a CRISPR/Cas9 binary vector (pBAtC) for both pepper types. Finally, we screened transformed calli for PPT resistance (1 and 5 mg/L PPT for cv. CM334 and Dempsey, respectively). These selected calli showed different indel frequencies from the non-transformed calli. However, the primary indel pattern was consistent with a 1-bp deletion at the target locus of the C. annuumMLO gene (CaMLO2). These results demonstrate the different sensitivity between cv. CM334 and Dempsey to A. tumefaciens-mediated callus induction, and a differential selection pressure of PPT via pBAtC binary vector.

A wireless closed-loop system for optogenetic peripheral neuromodulation.

The fast-growing field of bioelectronic medicine aims to develop engineered systems that can relieve clinical conditions by stimulating the peripheral nervous system1-5. This type of technology relies largely on electrical stimulation to provide neuromodulation of organ function or pain. One example is sacral nerve stimulation to treat overactive bladder, urinary incontinence and interstitial cystitis (also known as bladder pain syndrome)4,6,7. Conventional, continuous stimulation protocols, however, can cause discomfort and pain, particularly when treating symptoms that can be intermittent (for example, sudden urinary urgency)8. Direct physical coupling of electrodes to the nerve can lead to injury and inflammation9-11. Furthermore, typical therapeutic stimulators target large nerve bundles that innervate multiple structures, resulting in a lack of organ specificity. Here we introduce a miniaturized bio-optoelectronic implant that avoids these limitations by using (1) an optical stimulation interface that exploits microscale inorganic light-emitting diodes to activate opsins; (2) a soft, high-precision biophysical sensor system that allows continuous measurements of organ function; and (3) a control module and data analytics approach that enables coordinated, closed-loop operation of the system to eliminate pathological behaviours as they occur in real-time. In the example reported here, a soft strain gauge yields real-time information on bladder function in a rat model. Data algorithms identify pathological behaviour, and automated, closed-loop optogenetic neuromodulation of bladder sensory afferents normalizes bladder function. This all-optical scheme for neuromodulation offers chronic stability and the potential to stimulate specific cell types.

Flexible Near-Field Wireless Optoelectronics as Subdermal Implants for Broad Applications in Optogenetics.

In vivo optogenetics provides unique, powerful capabilities in the dissection of neural circuits implicated in neuropsychiatric disorders. Conventional hardware for such studies, however, physically tethers the experimental animal to an external light source, limiting the range of possible experiments. Emerging wireless options offer important capabilities that avoid some of these limitations, but the current size, bulk, weight, and wireless area of coverage is often disadvantageous. Here, we present a simple but powerful setup based on wireless, near-field power transfer and miniaturized, thin, flexible optoelectronic implants, for complete optical control in a variety of behavioral paradigms. The devices combine subdermal magnetic coil antennas connected to microscale, injectable light-emitting diodes (LEDs), with the ability to operate at wavelengths ranging from UV to blue, green-yellow, and red. An external loop antenna allows robust, straightforward application in a multitude of behavioral apparatuses. The result is a readily mass-producible, user-friendly technology with broad potential for optogenetics applications.

Polymer-hybridized liposomes of poly(amino acid) derivatives as transepidermal carriers.

This work describes the use of a novel transepidermal drug carrier system composed of phospholipids and amphiphilc poly(amino acid)s. We polymerized poly(asparagine) grafted with octadecylamine (PAsn-g-C18), poly(aspartic acid) grafted with octadecylamine (PAsp-g-C18), and poly(aspartic acid) grafted with phytosphingosine (PAsp-g-PHS). We then prepared polymer hybridized liposomes (PHL) anchored with alkyl grafted poly(amino acid)s and encapsulated hydrolyzed ginseng saponins (HGS). We confirmed that the liposomes and PHL reduce the cytotoxicity of HGS, which was not observed with polymeric nano-carriers. A quantitative analysis of the amount of penetrated HGS using the Franz cell method revealed that skin permeation of the lipophilic drugs loaded in liposomes was enhanced by the incorporation of amphiphilic poly(amino acid)s. Fluorescence microscopy observations also demonstrated excellent skin permeation performance of PHL anchored with PAsp-g-PHS. PHL showed better structural stability than liposomes in an O/W emulsion. PHL considerably improved the chemical stability of HGS compared to the liposomes. It is thought that the skin permeability of encapsulated bioactive molecules could be affected by the vesicle structure, membrane fluidity, and the type of anchored polymer.

Canted antiferromagnetism and spin reorientation transition in layered inorganic-organic perovskite (C6H5CH2CH2NH3)2MnCl4.

This work presents the synthesis, structure determination and magnetic properties of a new complex, phenethylammonium tetrachloromanganate(II), (C(6)H(5)CH(2)CH(2)NH(3))(2)MnCl(4) (Mn-PEA). Single crystals of Mn-PEA were obtained from methanol solution using the solvent-evaporation method at room temperature. The crystal structure of Mn-PEA was determined by single-crystal X-ray diffraction (orthorhombic, space group Pbca, a = 7.2075(9), b = 7.3012(14), c = 39.413(6) Å and Z = 4). The structure consisted of an extended [MnCl(4)](2-) network and two phenethylammonium cations to form a two-dimensional halide perovskite structure. Temperature-dependent magnetization measurements indicated that Mn-PEA acted as a weak ferromagnet below T(C) = 44.3 K due to spin canting. Below T(C), the magnetic behavior differed significantly from the behavior commonly observed among weak ferromagnets. The susceptibility depended strongly on the crystal orientation, the external magnetic field strength, and the magnetic history. The isothermal magnetization for two orientations revealed a ferromagnetic moment with a spin-canting angle of 0.04° and a spin-flop transitions with H(sf) = 3.5 T. The weak ferromagnetism, which manifested as spontaneous magnetization and magnetic hysteresis near a field strength of zero, was driven by interplay between the easy axis and the antisymmetric Dzyaloshinsky-Moriya (DM) interaction, leading to directional dependent magnetic behavior.

Cross-linked magnetic nanoparticles from poly(ethylene glycol) and dodecyl grafted poly(succinimide) as magnetic resonance probes.

Cross-linked magnetic nanoparticles were developed to improve the structural stability of amphiphilic polymer coated magnetic nanoparticles. These nanoparticles show strong potential for biomedical applications such as magnetic resonance imaging (MRI).

Polymer-hybridized liposomes anchored with alkyl grafted poly(asparagine).

Polymer-hybridized liposomes (PHLs) of saturated lecithin were formed by association of poly(asparagines) grafted with alkyl chains (PAsn-g-Cn). The thermal, physical, and surface properties of the polymer-hybridized liposomes were examined with varying polymer concentration, alkyl chain length (C(8), C(12), C(18), C(22)), and degree of substitution (DS) in the polymer. The inclusion of the polymer raised the membrane fluidity of liposomes. By the incorporation of small amount of polymer, the membrane rigidity of liposomes dropped sharply and then increased close to the original level as the polymer concentrations increased in the cases of PAsn-g-C(18) and PAsn-g-C(22). Also, the membrane rigidity and stability of PHLs increased with alkyl chain length at the same polymer concentration. The surface charge of PHL associated with PAsn-g-C(22) was changed by DS of alkyl chains. The polymer bearing long alkyl chains (C(12), C(18), C(22)) formed PHLs well at low polymer concentration and the number of disk-shaped polymer-lipid mixed micelles increased with polymer concentration. The anchored polymers induced shifts in gel-to-liquid crystal transition temperature (Tc) of the vesicles and Tc varied with polymer concentration, alkyl chain length, and DS of the polymer.

Incidental findings of the lumbar spine at MRI during herniated intervertebral disk disease evaluation.

OBJECTIVE: The objective of our study was to evaluate the frequency and types of incidental findings of the lumbar spine during MR evaluation for herniated intervertebral disk disease. MATERIALS AND METHODS: A total of 1268 patients (male-to-female ratio, 421:847; age range, 1-97 years) with clinically suspected herniated intervertebral disk disease underwent MRI of the lumbar spine. Musculoskeletal radiologists evaluated the MR examinations for the presence of incidental findings. We defined incidental finding as any abnormal finding not related to the chief complaint. Vertebral hemangioma, Tarlov cyst, fibrolipoma, synovial cyst, and sacral meningocele were included. Frequency distributions of the assessed imaging characteristics were calculated. For analysis of the relationship of incidental findings with patient characteristics, the chi-square test was used. RESULTS: Overall, 107 patients (8.4%) had incidental findings. Fibrolipoma was most common (41 cases, 3.2%), followed by Tarlov cyst (27 cases, 2.1%) and vertebral hemangioma (19 cases, 1.5%). Fibrolipoma and sacral meningocele were more common in males (p < 0.05). There was no difference in the incidence between the sexes in the other incidental findings (p = 0.26-0.96). Four of the five incidental findings were significantly more frequent in individuals younger than 50 years (p < 0.05), whereas the incidence of vertebral hemangioma did not differ by patient age (p = 0.32). CONCLUSION: Incidental findings at MRI of the lumbar spine were common and associated with age and sex. Most were benign findings. An awareness of the prevalence of the incidental findings detected at MRI of the lumbar spine is helpful for diagnosing lesions not related to symptoms.

Gray-scale and color Doppler sonographic appearances of nonsubungual soft-tissue glomus tumors.

BACKGROUND: To evaluate the sonographic findings of soft-tissue nonsubungual glomus tumors. METHODS: The sonographic appearances of nine histologically proven soft-tissue glomus tumors of nonsubungual location in nine patients (mean age, 49 years; M:F = 7:2) were reviewed retrospectively. Doppler examination and surgical excision were performed in all cases. RESULTS: The mean size of the lesions was 1 cm. The margins of the lesions were relatively well-circumscribed in eight of nine patients (89%) with an ovoid shape in seven of nine patients (78%). The vascularity was moderate to rich in all cases, with an arterial flow pattern but no arteriovenous shunt patterns. The "vascular stalk sign" was noted in six cases (67%). CONCLUSIONS: Nonsubungual glomus tumors are rare soft-tissue tumors with abundant vascularity and arterial flow pattern.

Sonographic findings in Nicolau syndrome following intramuscular diclofenac injection: a case report.

Nicolau syndrome (also known as embolia cutis medicamentosa and livedoid dermatitis) is a rare but severe localized adverse drug reaction of intramuscular injection of various drugs. The typical presentation is pain around the injection site soon after injection, followed by erythema, livedoid patch, hemorrhagic patch, and finally necrosis of skin, subcutaneous fat, and muscle tissue. We report a case that occurred in a 34-year-old woman after intramuscular injection of diclofenac sodium. Sonography showed diffuse thickening with increased echogenicity of the skin and subcutaneous fat layer, while MRI revealed extensive edema involving gluteal and piriformis muscles and deep fascia, and fluid collection.

Preoperative localization of cystic lesions in the knee using ultrasound-guided injection of indigo carmine.

PURPOSE: To evaluate the feasibility and effectiveness of preoperative localization of cystic lesions in the knee using ultrasound-guided indigo carmine injection. METHOD: Twenty-three cysts in the knee in 23 patients (M:F = 15:8, mean age, 42 years) were localized preoperatively by ultrasound-guided indigo carmine injection. These included 12 meniscal cysts, 7 popliteal cysts, and 4 ganglion cysts. To stain the lesions, 0.2-3 mL of indigo carmine was injected into the cyst using a 22-gauge spinal needle. After localization, the patient was immediately transferred to the operating room and surgery was performed. Intraoperative findings and arthroscopic images were reviewed. RESULT: All 23 cysts were stained successfully. Twenty cases were confirmed during arthroscopy and 3 cases were confirmed during excisional surgery. There was no significant bleeding/hematoma or anaphylactic reaction. Four patients felt pain during aspiration before indigo carmine injection. The lesions were stained blue and could be clearly identified by the surgeon and were removed arthroscopically or by open surgery. CONCLUSION: Preoperative localization of cystic lesions in the knee joint region using ultrasound-guided indigo carmine injection is a feasible technique and can be easily and safely be performed.

CD40 ligation of rheumatoid synovial fibroblasts regulates RANKL-mediated osteoclastogenesis: evidence of NF-kappaB-dependent, CD40-mediated bone destruction in rheumatoid arthritis.

OBJECTIVE: To determine whether CD40 ligation of rheumatoid arthritis synovial fibroblasts (RASFs) is able to induce RANKL expression and osteoclastogenesis in RASFs, and to identify its mechanism of action in patients with RA. METHODS: CD40 of RASFs was ligated with CD40 ligand (CD40L)-transfected L cells or activated T cells. The formation of osteoclasts in cocultures of CD40-ligated RASFs and T lymphocyte-depleted peripheral blood mononuclear cells was evaluated by tartrate-resistant acid phosphatase staining, detection of calcitonin receptor, and resorption pit formation assay. The expression of NF-kappaB, IkappaB alpha, ERK-1/2, phospho-ERK-1/2, p38, phospho-p38, and RANKL was examined by immunoblotting and/or semiquantitative reverse transcription-polymerase chain reaction. RESULTS: CD40 ligation of RASFs by CD40L-transfected L cells or activated T cells induced RANKL expression and enhanced osteoclastogenesis. CD40 ligation of RASFs also induced activation of ERK-1/2, p38 MAPK, and NF-kappaB and up-regulation of CD40 ligation-induced RANKL expression, whereas osteoclastogenesis was reduced in RASFs transfected with a dominant-negative mutant of IkappaB alpha or by an NF-kappaB inhibitor. However, specific inhibitors of MAPK/ERK-1/2 and p38 MAPK partially blocked the induction of RANKL expression and osteoclastogenesis. Monoclonal antibodies against interleukin-1 and tumor necrosis factor alpha partially inhibited CD40 ligation-mediated osteoclastogenesis. CONCLUSION: These results indicate that CD40 ligation of RASFs induces RANKL expression mainly via NF-kappaB activation and also results in enhanced osteoclast formation, both of which might play important roles in bone and cartilage destruction in RA. Inhibition of the CD40-CD40L interaction is a potential strategy for the prevention of bone damage in RA.

The flavonoid ellagic acid from a medicinal herb inhibits host immune tolerance induced by the hepatitis B virus-e antigen.

The aim of this study is to characterize the role of ellagic acid, a flavonoid from a medicinal herb which blocks HBeAg secretion in a HBV infected cell line and in HBeAg transgenic mice, in immune tolerance in chronic HBV infection. Using the mouse strain C57ML/6, HBeAg-producing transgenic mice (HBeAg-Tg), under the control of metal ion-inducible promoter were generated. The effect on immune tolerance of HBeAg-Tg and the release of immune tolerance by the inhibitor of HBeAg secretion, ellagic acid, was tested using T/B cell proliferation, HBeAg/HBeAb production, cytotoxic T-lymphocyte (CTL) and cytokine assays. C57ML/6 based HBeAg-producing HBeAg-Tg mice were tolerant to HBeAg at the T and B-cell level, did not produce antibodies to HBeAg in vivo and in vitro, produced minimal levels of cytokines (IL-4 and IFN-gamma) and decreased CTL responses, while feeding mice with ellagic acid (5mg/kg body weight) blocked the immune tolerance caused by HBeAg. Our results suggest that host immune tolerance induced by HBeAg during HBV infection, a viral strategy to guarantee HBV infection, can be overcome by ellagic acid, thus it can be used as a therapeutic for HBV-carriers.

Smectic ordering in liquid-crystal-aerosil dispersions. II. Scaling analysis.

Liquid crystals offer many unique opportunities to study various phase transitions with continuous symmetry in the presence of quenched random disorder (QRD). The QRD arises from the presence of porous solids in the form of a random gel network. Experimental and theoretical work supports the view that for fixed (static) inclusions, quasi-long-range smectic order is destroyed for arbitrarily small volume fractions of the solid. However, the presence of porous solids indicates that finite-size effects could play some role in limiting long-range order. In an earlier work, the nematic-smectic-A transition region of octylcyanobiphenyl (8CB) and silica aerosils was investigated calorimetrically. A detailed x-ray study of this system is presented in the preceding paper, which indicates that pseudocritical scaling behavior is observed. In the present paper, the role of finite-size scaling and two-scale universality aspects of the 8CB+aerosil system are presented and the dependence of the QRD strength on the aerosil density is discussed.