Development of an indirect ELISA system for diagnosis of porcine edema disease using recombinant modified Stx2e antigen.

AIM: This study aimed to develop a sensitive and specific recombinant antigen protein indirect enzyme-linked immunosorbent assay (ELISA) kit to detect the Shiga toxin (Stx)-producing Escherichia coli (STEC) antibodies against porcine edema disease (ED). METHODS AND RESULTS: The recombinant antigen was co-expressed with the STEC-derived Stx2e A2-fragment and Stx2e B protein in E. coli BL21(DE3) pLysS cells and purified using maltose-binding protein open columns. We used a Shiga-like toxin 2 antibody to test the specificity of the recombinant antigen in an indirect ELISA, which was detected in antigen-coated wells but not in uncoated wells. We tested the indirect ELISA system using samples from the STEC-immunized pig group, the commercial swine farm group, and healthy aborted fetal pleural effusion group; five and twenty samples, respectively, were positive for STEC in the former, whereas all three samples were negative for STEC in the latter. CONCLUSIONS: This newly developed indirect ELISA may be a specific method for diagnosing STEC infections in pigs.

Development of a Novel Short Synthetic Antibacterial Peptide Derived from the Swallowtail Butterfly Papilio xuthus Larvae.

Insects possess biological defense systems that can effectively combat the invasion of external microorganisms and viruses, thereby supporting their survival in diverse environments. Antimicrobial peptides (AMPs) represent a fast-acting weapon against invading pathogens, including various bacterial or fungal strains. A 37-residue antimicrobial peptide, papiliocin, derived from the swallowtail butterfly Papilio xuthus larvae, showed significant antimicrobial activities against several human pathogenic bacterial and fungal strains. Jelleines, isolated as novel antibacterial peptides from the Royal Jelly (RJ) of bees, exhibit broad-spectrum protection against microbial infections. In this study, we developed a novel antimicrobial peptide, PAJE (RWKIFKKPFKISIHL-NH2), which is a hybrid peptide prepared by combining 1-7 amino acid residues (RWKIFKK-NH2) of papiliocin and 1-8 amino acid residues (PFKISIHL-NH2) of Jelleine-1 to alter length, charge distribution, net charge, volume, amphipaticity, and improve bacterial membrane interactions. This novel peptide exhibited increased hydrophobicity and net positive charge for binding effectively to the negatively charged membrane. PAJE demonstrated antimicrobial activity against both gram-negative and gram-positive bacteria, with very low toxicity to eukaryotic cells and an inexpensive process of synthesis. Collectively, these findings suggest that this novel peptide possesses great potential as an antimicrobial agent.

Development of a real-time loop-mediated isothermal amplification method for the detection of severe fever with thrombocytopenia syndrome virus.

Severe fever with thrombocytopenia syndrome (SFTS) is being reported annually in South Korea since its first detection there in 2010. The causal agent is a negative-strand RNA virus 80-100 nm in diameter. It causes fever, thrombocytopenia, leukocytopenia, gastrointestinal symptoms, and neural symptoms. The mortality rate of SFTS was 32.6% among 172 cases reported from 2012 to 2015 in South Korea. Thus, is necessary to develop an effective diagnostic method that selectively identifies the isolates circulating in South Korea. The real-time reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay is a simple, rapid, and sensitive approach for molecular diagnosis. Here, we designed novel primers for this assay and found that the technique had very high specificity, sensitivity, and efficiency. This real-time RT-LAMP approach using the novel primers developed herein can be applied for early diagnosis of SFTSV strains in South Korea to reduce the mortality rate of SFTS.

Risk Factors for Symptomatic Contralateral Foraminal Stenosis After Unilateral Transforaminal Lumbar Interbody Fusion.

OBJECTIVE: The most common cause of contralateral symptoms after unilateral transforaminal lumbar interbody fusion (TLIF) is contralateral foraminal stenosis (FS). This retrospective cohort study aimed to investigate the cause of and risk factors for contralateral FS after unilateral TLIF with a single cage. METHODS: Patients with degenerative lumbar spinal disorders who underwent unilateral TLIF at L4-5 were divided into 2 groups: those without contralateral radicular symptoms after surgery (group A; n = 340) and those with contralateral radicular symptoms after surgery (group B; n = 16). We investigated the influence of various radiological and cage-related factors on postoperative contralateral FS with radicular symptoms. The cage location indicates whether the cage's anterior tip crosses the disc midline-exceeding 50%-and in such a case, how far. RESULTS: Group B showed significantly increased postoperative coronal angle and sagittal angle and decreased contralateral foraminal height and foraminal area. Statistically significant (P < 0.01) factors according to the multivariate logistic regression analysis were the preoperative sagittal range of motion (odds ratio [OR]: 1.562, P = 0.004) and cage location (OR: 2.047, P = 0.015). The cutoff values for the sagittal range of motion and the cage location were 9.0° and 50.5%, respectively. The preoperative and postoperative 6-month visual analog scale scores and Oswestry disability index values were not significantly different between the groups. CONCLUSIONS: The 2 most meaningful risk factors were the preoperative sagittal range of motion and cage location. Inserting the cage beyond the disc midline, especially in patients with a high preoperative sagittal range of motion (≥9.0°), would help reduce postoperative complications.

Hydrogen Production by Steam Reforming of Natural Gas Over Vanadium-Nickel-Alumina Catalysts.

A series of vanadium-nickel-alumina (xVNA) catalysts were prepared by a single-step sol-gel method with a variation of vanadium content (x, wt%) for use in the hydrogen production by steam reforming of natural gas. The effect of vanadium content on the physicochemical properties and catalytic activities of xVNA catalysts in the steam reforming of natural gas was investigated. It was found that natural gas conversion and hydrogen yield showed volcano-shaped trends with respect to vanadium content. It was also revealed that natural gas conversion and hydrogen yield increased with decreasing nickel crystallite size.

Genome sequence of the Japanese oak silk moth, Antheraea yamamai: the first draft genome in the family Saturniidae.

Background: Antheraea yamamai, also known as the Japanese oak silk moth, is a wild species of silk moth. Silk produced by A. yamamai, referred to as tensan silk, shows different characteristics such as thickness, compressive elasticity, and chemical resistance compared with common silk produced from the domesticated silkworm, Bombyx mori. Its unique characteristics have led to its use in many research fields including biotechnology and medical science, and the scientific as well as economic importance of the wild silk moth continues to gradually increase. However, no genomic information for the wild silk moth, including A. yamamai, is currently available. Findings: In order to construct the A. yamamai genome, a total of 147G base pairs using Illumina and Pacbio sequencing platforms were generated, providing 210-fold coverage based on the 700-Mb estimated genome size of A. yamamai. The assembled genome of A. yamamai was 656 Mb (>2 kb) with 3675 scaffolds, and the N50 length of assembly was 739 Kb with a 34.07% GC ratio. Identified repeat elements covered 37.33% of the total genome, and the completeness of the constructed genome assembly was estimated to be 96.7% by Benchmarking Universal Single-Copy Orthologs v2 analysis. A total of 15 481 genes were identified using Evidence Modeler based on the gene prediction results obtained from 3 different methods (ab initio, RNA-seq-based, known-gene-based) and manual curation. Conclusions: Here we present the genome sequence of A. yamamai, the first genome sequence of the wild silk moth. These results provide valuable genomic information, which will help enrich our understanding of the molecular mechanisms relating to not only specific phenotypes such as wild silk itself but also the genomic evolution of Saturniidae.

Bombyx mori hemocyte extract has anti-inflammatory effects on human phorbol myristate acetate-differentiated THP­1 cells via TLR4-mediated suppression of the NF-κB signaling pathway.

Hemolymph is the circulating fluid of insects and is a key component of their immune system. However, little is known concerning hemocyte identification, development, differentiation and related cellular immune responses. The present study aimed to determine whether a hemocyte extract prepared from Bombyx mori larvae had anti­inflammatory effects; THP­1 (a human monocytic leukemia cell line) cells that had been differentiated into macrophage­like cells by treatment with phorbol myristate acetate (PMA) were used. THP­1 cells were cultured with different concentrations of a B. mori hemocyte extract prior to exposure to lipopolysaccharide (LPS) to induce an inflammatory response. The effects of the B. mori hemocyte extract on anti­inflammatory pathways were determined using reverse transcription­quantitative polymerase chain reaction and western blotting to assess the expression of pro­inflammatory molecules. The B. mori hemocyte extract inhibited the LPS­induced mRNA expression of Toll­like receptor 4 in addition to LPS­induced interleukin (IL)­1ß, IL­6, IL­8 and tumor necrosis factor­α. Treatment of PMA­differentiated THP­1 cells with B. mori hemocyte extract also inhibited inducible nitric oxide synthase and cyclooxygenase­2 transcription and translation. Nuclear factor­κB activation and phosphorylation also decreased. Further in­depth functional studies are required to understand the mechanism underlying the anti­inflammatory effects of silkworm hemocyte extract.

Central Administration of 1-Deoxynojirimycin Attenuates Hypothalamic Endoplasmic Reticulum Stress and Regulates Food Intake and Body Weight in Mice with High-Fat Diet-Induced Obesity.

The α-glucosidase inhibitor, 1-deoxynojirimycin (DNJ), is widely used for its antiobesity and antidiabetic effects. Researchers have demonstrated that DNJ regulates body weight by increasing adiponectin levels, which affects energy intake and prevents diet-induced obesity. However, the mechanism by which centrally administered DNJ exerts anorexigenic effects has not been studied until now. We investigated the effect of DNJ in the hypothalamus of mice with high-fat diet-induced obesity. Results showed that intracerebroventricular (ICV) administration of DNJ reduced hypothalamic ER stress, which activated the leptin-induced Janus-activated kinase 2 (JAK2)/signal transducers and activators of transcription 3 (STAT3) signaling pathway to cause appetite suppression. We conclude that DNJ may reduce obesity by moderating feeding behavior and ER stress in the hypothalamic portion of the central nervous system (CNS).

Preservation of Motion at the Surgical Level after Minimally Invasive Posterior Cervical Foraminotomy.

OBJECTIVE: Although minimally invasive posterior cervical foraminotomy (MI-PCF) is an established approach for motion preservation, the outcomes are variable among patients. The objective of this study was to identify significant factors that influence motion preservation after MI-PCF. METHODS: Forty-eight patients who had undergone MI-PCF between 2004 and 2012 on a total of 70 levels were studied. Cervical parameters measured using plain radiography included C2-7 plumb line, C2-7 Cobb angle, T1 slope, thoracic outlet angle, neck tilt, and disc height before and 24 months after surgery. The ratios of the remaining facet joints after MI-PCF were calculated postoperatively using computed tomography. Changes in the distance between interspinous processes (DISP) and the segmental angle (SA) before and after surgery were also measured. We determined successful motion preservation with changes in DISP of ≤3 mm and in SA of ≤2°. RESULTS: The differences in preoperative and postoperative DISP and SA after MI-PCF were 0.03±3.95 mm and 0.34±4.46°, respectively, fulfilling the criteria for successful motion preservation. However, the appropriate level of motion preservation is achieved in cases in which changes in preoperative and postoperative DISP and SA motions are 55.7 and 57.1%, respectively. Based on preoperative and postoperative DISP, patients were divided into three groups, and the characteristics of each group were compared. Among these, the only statistically significant factor in motion preservation was preoperative disc height (Pearson's correlation coefficient=0.658, p<0.001). The optimal disc height for motion preservation in regard to DISP ranges from 4.18 to 7.08 mm. CONCLUSION: MI-PCF is a widely accepted approach for motion preservation, although desirable radiographic outcomes were only achieved in approximately half of the patients who had undergone the procedure. Since disc height appears to be a significant factor in motion preservation, surgeons should consider disc height before performing MI-PCF.

Inhibin-α gene mutations and mRNA levels in human lymphoid and myeloid leukemia cells.

The inhibin-α gene was identified as a tumor suppressor gene in the gonads and adrenal glands by functional studies using knockout mice. Methylation of CpG sites within the regulatory regions of tumor suppressor gene is frequently associated with their transcriptional silencing. We investigated epigenetic modifications, changes in loss of heterozygosity (LOH), and mutation of the inhibin-α gene, and regulation of transcriptional expression in response to inhibitors of DNA methylation (5-aza-2'-deoxycytidine, 5-AzaC) in human lymphoid (Jurkat, Molt-4, Raji, and IM-9) and myeloid (HL-60, Kasumi-1, and K562) leukemia cells. The inhibin-α promoter was hypermethylated in lymphoid (Molt-4 and Raji) and myeloid (HL-60 and Kasumi-1) leukemia cells. Inhibin-α gene mutations differed significantly between lymphoid (heterozygote) and myeloid (homozygote) leukemia cells. LOH in the inhibin-α gene was detected in lymphoid and myeloid leukemia cells, with the exception of Jurkat cells. Treatment with 5-AzaC, a demethylating agent, resulted in increased inhibin-α mRNA and protein levels in most of the cell lines. Also, 5-AzaC treatment inhibited cell proliferation and induced apoptosis. Taken together, our results reveal that the inhibin-α gene is transcriptionally silenced in human leukemia cells and that reactivation is suppressed by a demethylating agent. In addition, mutations in, and expression levels of, the inhibin-α gene differed between human lymphoid and myeloid leukemia cells.

Hydrogen Production by Steam Reforming of Liquefied Natural Gas (LNG) Over Nickel-Phosphorus-Alumina Xerogel Catalyst Prepared by a Carbon-Templating Epoxide-Driven Sol-Gel Method.

A nickel-phosphorus-alumina xerogel catalyst was prepared by a carbon-templating epoxide-driven sol-gel method (denoted as CNPA catalyst), and it was applied to the hydrogen production by steam reforming of liquefied natural gas (LNG). For comparison, a nickel-phosphorus-alumina xerogel catalyst was also prepared by a similar method in the absence of carbon template (denoted as NPA catalyst). The effect of carbon template addition on the physicochemical properties and catalytic activities of the catalysts in the steam reforming of LNG was investigated. Both CNPA and NPA catalysts showed excellent textural properties with well-developed mesoporous structure. However, CNPA catalyst retained a more reducible nickel aluminate phase than NPA catalyst. XRD analysis of the reduced CNPA and NPA catalysts revealed that nickel sintering on the CNPA catalyst was suppressed compared to that on the NPA catalyst. From H2-TPD and CH4-TPD measurements of the reduced CNPA and NPA catalysts, it was also revealed that CNPA catalyst with large amount of hydrogen uptake and strong hydrogen-binding sites showed larger amount of methane adsorption than NPA catalyst. In the hydrogen production by steam reforming of LNG, CNPA catalyst with large methane adsorption capacity showed a better catalytic activity than NPA catalyst.

The Prevalence of Undiagnosed Presurgical Cognitive Impairment and Its Postsurgical Clinical Impact in Older Patients Undergoing Lumbar Spine Surgery.

OBJECTIVE: Because elderly patients are undergoing more surgeries, the importance of postoperative cognitive impairment (CI) evaluations is rising, especially for spine surgery, which is related to subjective pain. We investigated the prevalence of undiagnosed CI among elderly patients who underwent spine surgery and the impact of CI on postoperative outcomes. METHODS: The preoperative cognitive statuses of 129 patients over 65 who underwent lumbar spine surgery from 2012 to 2014 were determined with the Mini-Mental State Examination, and patients with scores under 24 were diagnosed with CI. The patients were then divided into a CI group (n=49) and non-cognitive impairment (NCI) group (n=80). RESULTS: Among the 129 patients, 49 (38.0%) were diagnosed with CI, and 9 (7.0%) had severe CI. The age of the CI group (72.88±6.20 years) was significantly greater than that of the NCI group (69.96±4.53 years). In contrast, the postoperative visual analog scale scores and performance statuses did not differ significantly. However, postoperative delirium was more frequent and the hospital stay length was longer in the CI group compared with the NCI group (p<0.05). CONCLUSION: A high prevalence of undiagnosed CI was discovered among elderly patients undergoing spine surgery. The existence of CI was associated with higher rates of postoperative delirium and prolonged hospital stays, which affected clinical outcomes. Thus, CI assessments should be included in preoperative evaluations of elderly patients prior to spine surgery.

Spinous Process-Splitting Hemilaminoplasty for Intradural and Extradural Lesions.

OBJECTIVE: To describe a novel spinous process-splitting hemilaminoplasty technique for the surgical treatment of intradural and posterior epidural lesions that promotes physiological restoration. METHODS: The spinous process was split, the area of the facet lamina junction was drilled, and en bloc hemilaminectomy was then performed. After removing intradural and posterior epidural lesions, we fitted the previously en bloc-removed bone to the pre-surgery same shape, and held it in place with non-absorbable sutures. Surgery was performed on 16 laminas from a total of nine patients between 2011 and 2014. Bony union of the reconstructed lamina was assessed using computed tomography (CT) at 6 months after surgery. RESULTS: Spinous process-slitting hemilaminoplasty was performed for intradural extramedullary tumors in eight patients and for ossification of the ligament flavum in one patient. Because we were able to visualize the margin of the ipsilateral and contralateral dura, we were able to secure space for removal of the lesion and closure of the dura. None of the cases showed spinal deformity or other complications. Bone fusion and maintenance of the spinal canal were found to be perfect on CT scans. CONCLUSION: The spinous process-splitting hemilaminoplasty technique presented here was successful in creating sufficient space to remove intradural and posterior epidural lesions and to close the dura. Furthermore, we were able to maintain the physiological barrier and integrity after surgery because the posterior musculature and bone structures were restored.

Acute Contralateral Radiculopathy after Unilateral Transforaminal Lumbar Interbody Fusion.

OBJECTIVE: Cases of contralateral radiculopathy after a transforaminal lumbar interbody fusion with a single cage (unilateral TLIF) had been reported, but the phenomenon has not been explained satisfactorily. The purpose of this study was to determine its incidence, causes, and risk factors. METHODS: We did retrospective study with 546 patients who underwent a unilateral TLIF, and used CT and MRI to study the causes of contralateral radicular symptoms that appeared within a week postoperatively. Clinical and radiological results were compared by dividing the patients into the symptomatic group and asymptomatic group. RESULTS: Contralateral symptoms occurred in 32 (5.9%) of the patients underwent unilateral TLIF. The most common cause of contralateral symptoms was a contralateral foraminal stenosis in 22 (68.8%), screw malposition in 4 (12.5%), newly developed herniated nucleus pulposus in 3 (9.3%), hematoma in 1 (3.1%), and unknown origin in 2 patients (6.3%). 16 (50.0%) of the 32 patients received revision surgery. There was no difference in visual analogue scale and Oswestry disability index between the two groups at discharge. Both preoperative and postoperative contralateral foraminal areas were significantly smaller, and postoperative segmental angle was significantly greater in the symptomatic group comparing to those of the asymptomatic group (p<0.05). CONCLUSION: The incidence rate is not likely to be small (5.9%). If unilateral TLIF is performed for cases when preoperative contralateral foraminal stenosis already exists or when a large restoration of segmental lordosis is required, the probability of developing contralateral radiculopathy is increased and caution from the surgeon is needed.

The Effects of Spinopelvic Parameters and Paraspinal Muscle Degeneration on S1 Screw Loosening.

OBJECTIVE: To investigate risk factors for S1 screw loosening after lumbosacral fusion, including spinopelvic parameters and paraspinal muscles. METHODS: We studied with 156 patients with degenerative lumbar disease who underwent lumbosacral interbody fusion and pedicle screw fixation including the level of L5-S1 between 2005 and 2012. The patients were divided into loosening and non-loosening groups. Screw loosening was defined as a halo sign larger than 1 mm around a screw. We checked cross sectional area of paraspinal muscles, mean signal intensity of the muscles on T2 weight MRI as a degree of fatty degeneration, spinopelvic parameters, bone mineral density, number of fusion level, and the characteristic of S1 screw. RESULTS: Twenty seven patients showed S1 screw loosening, which is 24.4% of total. The mean duration for S1 screw loosening was 7.3±4.1 months after surgery. Statistically significant risk factors were increased age, poor BMD, 3 or more fusion levels (p<0.05). Among spinopelvic parameters, a high pelvic incidence (p<0.01), a greater difference between pelvic incidence and lumbar lordotic angle preoperatively (p<0.01) and postoperatively (p<0.05). Smaller cross-sectional area and high T2 signal intensity in both multifidus and erector spinae muscles were also significant muscular risk factors (p<0.05). Small converging angle (p<0.001) and short intraosseous length (p<0.05) of S1 screw were significant screw related risk factors (p<0.05). CONCLUSION: In addition to well known risk factors, spinopelvic parameters and the degeneration of paraspinal muscles also showed significant effects on the S1 screw loosening.

Role of a Burr Hole and Calvarial Bone Marrow-Derived Stem Cells in the Ischemic Rat Brain: A Possible Mechanism for the Efficacy of Multiple Burr Hole Surgery in Moyamoya Disease.

OBJECTIVE: This study investigates the role of a burr hole and calvarial bone marrow-derived stem cells (BMSCs) in a transient ischemic brain injury model in the rat and postulates a possible mechanism for the efficacy of multiple cranial burr hole (MCBH) surgery in moyamoya disease (MMD). METHODS: Twenty Sprague-Dawley rats (250 g, male) were divided into four groups : normal control group (n=5), burr hole group (n=5), ischemia group (n=5), and ischemia+burr hole group (n=5). Focal ischemia was induced by the transient middle cerebral artery occlusion (MCAO). At one week after the ischemic injury, a 2 mm-sized cranial burr hole with small cortical incision was made on the ipsilateral (left) parietal area. Bromodeoxyuridine (BrdU, 50 mg/kg) was injected intraperitoneally, 2 times a day for 6 days after the burr hole trephination. At one week after the burr hole trephination, brains were harvested. Immunohistochemical stainings for BrdU, CD34, VEGF, and Doublecortin and Nestin were done. RESULTS: In the ischemia+burr hole group, BrdU (+), CD34 (+), and Doublecortin (+) cells were found in the cortical incision site below the burr hole. A number of cells with Nestin (+) or VEGF (+) were found in the cerebral parenchyma around the cortical incision site. In the other groups, BrdU (+), CD34 (+), Doublecortin (+), and Nestin (+) cells were not detected in the corresponding area. These findings suggest that BrdU (+) and CD34 (+) cells are bone marrow-derived stem cells, which may be derived from the calvarial bone marrow through the burr hole. The existence of CD34 (+) and VEGF (+) cells indicates increased angiogenesis, while the existence of Doublecortin (+), Nestin (+) cells indicates increased neurogenesis. CONCLUSION: Based on these findings, the BMSCs through burr holes seem to play an important role for the therapeutic effect of the MCBH surgery in MMD.

An Intramuscular Hemangioma at the Cervical Muscle: A Case Report.

Intramuscular hemangioma (IMH) primarily develops in the skeletal muscles of the limbs or trunk. The occurrence of IMH in the neck muscle is very rare. In the present report, we describe a case of IMH in the posterior neck muscle. A 58-year-old woman presented with a mass on the right posterior neck area, which had developed 2 years previously. The tumor was round in shape, had a size of 3.5×3.0 cm, and was fixed to the surrounding soft tissue. The patient complained of mild dull pain and tenderness at the mass without reddish discoloration. The mass showed a diurnal variation in size - the mass grew in size in the morning and became smaller in the afternoon. Preoperative study confirmed the vascular nature of the tumor. During peritumoral dissection, the mass shrank rapidly following profuse bleeding and arterial cauterization, and hence, it was difficult to distinguish the mass from the surrounding tissues. A total resection was possible with careful dissection of its fibrotic boundary. Based on the histological findings, a definitive diagnosis of cavernous type IMH was made. An initial suspicion of IMH, according to the clinical findings, would be helpful for decision making of further evaluations and surgical plan.

CSF flow pathways through the ventricle-cistern interfaces in kaolin-induced hydrocephalus rats-laboratory investigation.

PURPOSE: The goal of this study was to identify direct cerebrospinal fluid (CSF) pathways in the interface between ventricles and cisterns. Such routes are hypothesized to be involved in alternative CSF flows in abnormal circumstances of CSF circulation. METHODS: Chronic obstructive hydrocephalus models were induced in ten Sprague-Dawley rats with kaolin injection into the cisterna magna. Three weeks after the kaolin injection, when thick arachnoid fibrosis obliterated the fourth ventricular outlets, cationized ferritin was stereotactically infused as a tracer into the lateral ventricle in order to observe the pathways from the ventricles to the subarachnoid space. Animals were killed in 48 h and brains were sectioned. CSF flow pathways were traced by the staining of ferritin with ferrocyanide. RESULTS: Eight out of ten rats developed hydrocephalus. The subarachnoid membranes of the convexity and basal cisterns were severely adhered such that most of the ferritin remained in the ventricles whereas basal and convexity cisterns were clear of ferritin. In six out of the eight hydrocephalus rats, ferritin leaked from the third ventricle into the quadrigeminal cistern, and from the lateral ventricle into the ambient cistern. CONCLUSIONS: The interfaces between the third ventricle and the quadrigeminal cistern, and between the lateral ventricle and the ambient cistern appear to be alternative CSF pathways in a pathologic condition such as obstructive hydrocephalus.

A novel splice variant of the decapentaplegic (dpp) gene in the wild silkworm, Bombyx mandarina.

Decapentaplegic (dpp) is a member of the transforming growth factor-ß superfamily. Although the dpp gene and related pathways are known to play important roles in insect development, few studies have examined its function in Bombyx mori and Bombyx mandarina. To date, there have been no previous reports on novel splice variants of dpp in silkworm. In the present study, we conducted RT-PCR to examine dpp expression in the mid-gut tissue of B. mandarina and discovered a novel dpp isoform. The isoform sequence was confirmed using sequencing analysis and found to have 333 bp deletion compared to full-length cDNA encoding dpp. The deleted sequence encodes a region of the latency associated peptide (LAP) region of transforming growth factor-ß (TGF-ß), which may affect the activity and specificity of TGF-ß. Using variant calling analyses, we detected 7 candidate single nucleotide variants (SNVs) for different alternative splicing in dpp. This is the first report of a novel splice variant of the dpp gene in B. mandarina and these results provide insight about the domestication process and distinct phenotypic traits of B. mori and B. mandarina.

Activin suppresses LPS-induced Toll-like receptor, cytokine and inducible nitric oxide synthase expression in normal human melanocytes by inhibiting NF-κB and MAPK pathway activation.

Activins are dimeric growth and differentiation factors that belong to the transforming growth factor (TGF)-ß superfamily of structurally related signaling proteins. In the present study, we examined the mechanisms through which activin regulates the lipopolysaccharide (LPS)-induced transcription of Toll-like receptors (TLRs), cytokines and inducible nitric oxide synthase (iNOS) in human melanocytes, as well as the involvement of nuclear factor (NF)-κB and mitogen-activated protein kinase (MAPK) signaling. Cell proliferation was analyzed by cell viability assay, mRNA expression was detected by RT-qPCR, and protein expression was measured by western blot analysis. LPS increased the mRNA expression of TLRs (TLR1-10) and cytokines [interleukin (IL)-1ß, IL-6, IL-8 and TNF-α], as well as the mRNA and protein expression of iNOS. Activin decreased the LPS-induced TLR and cytokine mRNA expression, as well as the LPS-induced iNOS mRNA and protein expression. In addition, activin suppressed NF-κB p65 activation and blocked inhibitor of NF-κB (IκBα) degradation in LPS-stimulated melanocytes, and reduced LPS-induced p38 MAPK and MEK/ERK activation. On the whole, our results demonstrated that activin inhibited TLR and cytokine expression in LPS-activated normal human melanocytes and suppressed LPS-induced iNOS gene expression. Moreover, the anti-inflammatory effects of activin were shown to be mediated through the suppression of NF-κB and MAPK signaling, resulting in reduced TLR and iNOS expression, and in the inhibition of inflammatory cytokine expression.