Phthalyl starch nanoparticles as prebiotics enhanced nisin production in Lactococcus lactis through the induction of mild stress in probiotics.

AIM OF THE STUDY: Effect of internalized phthalyl starch nanoparticles (PSNs) on the antimicrobial ability of Lactococcus lactis (LL) KCTC 2013. METHODS AND RESULTS: Phthalyl starch nanoparticles were prepared by self-assembly of phthalyl starch and the amount of the hydrophobic phthalic moieties were characterized by nuclear magnetic resonance: PSN1 (DS: 14·3 mol.%), PSN2 (DS: 17·8 mol.%) and PSN3 (DS: 30·4 mol.%). The sizes of PSN1, PSN2 and PSN3 measured by dynamic light scattering were 364·7, 248·4 and 213·4 nm, respectively, and the surface charges of PSNs measured by electrophoretic light scattering were negative charges and PSNs were spherical in shape according to scanning electron microscope. It was found that when PSNs were treated with LL, the PSNs were internalized into LL through nanoparticle size-, energy- and glucose transporter-dependent mechanisms. The internalization was confirmed by confocal laser scanning microscopy and fluorescence-activated cell sorting. Nisin was isolated and identified by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Also, more nisin was produced from PSNs-treated LL than untreated- or starch-treated LL. Co-culture assay and agar diffusion test were performed to test the antimicrobial ability. Antimicrobial ability against Gram-negative Escherichia coli k88, Salmonella gallinarum and Gram-positive Listeria monocytogenes of LL treated with PSNs was higher than that of untreated or starch-treated group. Finally, it was found that the expression level of stress response genes dnaK, dnaJ and groES was significantly higher in PSNs-treated groups compared with starch-treated group or LL alone. CONCLUSION: The internalization of PSNs into LL enhanced the production of nisin through mild intracellular stimulation, resulting in enhanced antimicrobial ability. SIGNIFICANCE AND IMPACT OF THE STUDY: This study shows the promising potential of PSNs as new prebiotics for increasing the production of nisin, thus demonstrating a new method for the biological production of such antimicrobial peptides.

Lactoferrin causes IgA and IgG2b isotype switching through betaglycan binding and activation of canonical TGF-ß signaling.

Lactoferrin (LF), a pleiotropic iron-binding glycoprotein, is known to modulate the humoral immune response. However, its exact role in Ig synthesis has yet to be elucidated. In this study, we investigated the effect of LF on Ig production by mouse B cells and its underlying mechanisms. LF, like transforming growth factor (TGF)-ß1, stimulated B cells to produce IgA and IgG2b, while downregulating other isotypes. Using limiting dilution analysis, LF was shown to increase the frequency of IgA-secreting B-cell clones. This was paralleled by an increase in Ig germ-line α (GLα) transcripts, indicating that LF plays a role as an IgA switch factor. Interestingly, LF directly interacted with betaglycan (TGF-ß receptor III, TßRIII) and in turn induced phosphorylation of TßRI and Smad3 through formation of the TßRIII/TßRII/TßRI complex, leading to IgA isotype switching. Peroral administration of LF increased intestinal/serum IgA production as well as number of IgA plasma cells in lamina propria. Finally, we found that LF has an adjuvant activity when nontoxigenic Salmonella typhimurium was inoculated perorally, conferring protection against intragastrical infection of toxigenic S. typhimurium. These results suggest that LF has an important effect on the mucosal/systemic IgA response and can contribute to protection against intestinal pathogens.

Role of reactive oxygen species in p53 activation during cisplatin-induced apoptosis of rat mesangial cells.

BACKGROUND AND OBJECTIVES: Nephrotoxicity is one of the main side effects of the anticancer drug cisplatin, and one of its main therapeutic limitations. It has been suggested that p53 activation plays important roles in renal cell injury by cisplatin. However, the mechanism of p53 activation by cisplatin is unclear. This study examined whether reactive oxygen species (ROS) production by cisplatin would be linked to p53 activation in rat mesangial cells. MATERIALS AND METHODS: Renal cells were incubated with cisplatin in the absence or presence of pifithrin-a (PFT), N-acetyl-cysteine (NAC), or dimethylthiourea (DMT). Cell viability was evaluated by 3-(4,5-dimethyl-2-thiazol yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Apoptosis was evaluated by caspase-3 activity and cleavage of poly (ADP-ribose) polymerase (PARP). The relative levels of ROS and p53 phosphorylation were determined by fluorometric assay and Western blot analysis, respectively. RESULTS: Cisplatin induced apoptotic cell death via caspase-3 activation and PARP cleavage, and also increased p53 activation and ROS production. The p53 inhibitor PFT inhibited cisplatin-induced apoptosis. NAC and DMT, two antioxidants, also inhibited cisplatin-induced apoptosis. Interestingly, NAC and DMT reduced ROS production and suppressed p53 activation in renal cells exposed to cisplatin. CONCLUSIONS: Our results suggest that the ability of cisplatin to induce apoptosis of rat mesangial cells requires ROS-dependent p53 activation, thus, supporting the potential therapeutic role of antioxidants in preventing the cisplatin nephrotoxicity.

Allogeneic haematopoietic SCT for natural killer/T-cell lymphoma: a multicentre analysis from the Asia Lymphoma Study Group.

Eighteen patients (men=14; women=4) with natural killer (NK)/T-cell lymphomas (CR1, N=9; CR2, N=7; PR, N=1; progressive disease, N=1) undergoing allogeneic haematopoietic SCT (HSCT) (myeloablative, N=14; reduced intensity, N=4) were analyzed. With a median follow-up of 20.5 months, the 5-year OS was 57% and 5-year EFS was 51%. The use of the SMILE regimen pre-HSCT was the most important positive prognostic indicator, resulting in significantly superior OS and EFS (P<0.01). Acute GVHD had a significant negative impact on OS (P=0.03). CR1 and CR2 patients had similar survivals, but all patients who were not transplanted in remission died. Chronic GVHD, International Prognostic Index, disease stage, primary sites of involvement, conditioning regimen and source of HSC did not affect survival. Although allogeneic HSCT leads to reasonable survival for NK/T-cell lymphoma patients, results need to be compared with those in patients receiving L-asparaginase-containing regimens. Novel prognostic models incorporating biomarkers, such as circulating EBV DNA, are needed to identify high-risk patients who may benefit from allogeneic HSCT.

Field tests of Poly(I:C) immunization with nervous necrosis virus (NNV) in sevenband grouper, Epinephelus septemfasciatus (Thunberg).

It was recently reported that Poly(I:C) immunization with live nervous necrosis virus (NNV) confers protection in sevenband grouper, Epinephelus septemfasciatus (Thunberg), from NNV infection. In the present study, we conducted field tests with sevenband grouper for the evaluation of Poly(I:C) immunization efficacy. In the first experiment, sevenband grouper were immunized with NNV followed by Poly(I:C) administration 7 weeks before natural occurrence of viral nervous necrosis (VNN). Survival rate of the naïve fish was 71.0%, whereas that of the immunized fish was 99.8%. In the second experiment, sevenband grouper were immunized 10 months before VNN occurrence and survival rate of the non-treated and vaccinated fish was 79.5% and 97.5%, respectively. In the third experiment, we administered Poly(I:C) to sevenband grouper at 20 days after natural occurrence of VNN. The survival rate of the non-treated fish was 9.8%, whereas that of fish administered Poly(I:C) was 93.7%. Based on these results, it was concluded that Poly(I:C) immunization conferred protection in fish against NNV infection in field tests and the protection lasted more than 10 months. Furthermore, even after occurrence of VNN, fish mortality could be reduced by Poly(I:C) administration and there was an unexpected curative effect on VNN-affected fish.

Comprehensive histologic analysis of interstitial lipolysis with the 1444 nm wavelength during a 3-month follow-up.

A number of near-infrared wavelengths have been proposed and studied for laser lipolysis, but the histologic evaluation of tissue response to laser lipolysis during long-term follow-up has been lacking. A 1444 nm Nd:YAG laser with better absorption in both fat and water has recently attracted attention. The present study was designed to investigate the comprehensive histopathology of 1444 nm Nd:YAG laser-assisted lipolysis at different energy levels during a 3-month follow-up. Laser lipolysis was performed on porcine fat tissue in vivo using a 1444 nm Nd:YAG laser (AccuSculpt®, Lutronic Corporation, Ilsan, Republic of Korea) and the total energies delivered interstitially to 10x10 cm² areas were 750 J, 1500 J, 2250 J, 3000 J, 3750 J, 4500 J, and 5250 J. Biopsy samples were taken and histologically analyzed immediately after biopsy and at 1, 2, 4, and 12 weeks postoperatively. With a fluence setting above 3000J/100 cm², inflammation was severe and remained by the 3-month follow-up, resulting in severe scarring of the fat tissue. Below this energy level, mild lobular inflammation in the early phase biopsy had resolved with no scarring by the 3-month follow-up. No histologic changes in the epidermis or dermal connective tissue were present. This study suggested that controlling the energy level is important for clinical applications of laser lipolysis with no significant complications.

The effect of a 1550 nm fractional erbium-glass laser in female pattern hair loss.

BACKGROUND: Female pattern hair loss (FPHL) is the most common cause of hair loss in women, and its prevalence increases with advancing age. Affected women may experience psychological distress and social withdrawal. A variety of laser and light sources have been tried for treatment of hair loss, and some success has been reported. OBJECTIVE: The purpose of this study was to determine the efficacy and safety of a 1550 nm fractional erbium-glass laser in treatment of female pattern hair loss. PATIENTS AND METHODS: Twenty eight ethnic South Korean patients with varying degrees of FPHL were enrolled in the study. Patients received ten treatments with a 1550 nm fractional Er:Glass Laser (Mosaic, Lutronic Co., Ltd, Seoul, South Korea) at 2-weeks intervals using the same parameters (5-10 mm tip, 6 mJ pulse energy, 800 spot/cm(2) density, static mode). Phototrichogram and global photographs were taken at baseline and at the end of laser treatment, and analysed for changes in hair density and hair shaft diameter. Global photographs underwent blinded review by three independent dermatologists using a 7-point scale. Patients also answered questionnaires assessing hair growth throughout the study. All adverse effects were reported during the study. RESULTS: Twenty seven patients completed a 5-month schedule of laser treatment. One patient was excluded during treatment due to occurrence of alopecia areata. At the initial visit, mean hair density was 100 ± 14/cm(2) , and mean hair thickness was 58 ± 12 µm. After 5 months of laser treatment, hair density showed a marked increase to 157 ± 28/cm(2) (P < 0.001), and hair thickness also increased to 75 ± 13 µm (P < 0.001). Global photographs showed improvement in 24 (87.5%) of the 27 patients. Two patients (7.4%) reported mild pruritus after laser treatment; however, these resolved within 2 h. CONCLUSION: A 1550 nm fractional erbium-glass laser irradiation may be an effective and safe treatment option for women with female pattern hair loss.

Monitoring of viruses in chum salmon (Oncorhynchus keta) migrating to Korea.

It is important to investigate the prevalence of salmonid pathogens because they can affect the amount of release of salmonid fry and the migration rate of adult salmonids. In this study, routine surveys were conducted for investigating virus distribution in migrating chum salmon spawners (Oncorhynchus keta) and their offsprings at the Namdae River, Yangyang, Korea, during 2006-2008. Anterior kidneys were removed from chum salmon spawner individuals, homogenized with minimal essential medium, and centrifuged to make supernatants for conducting RT-PCR. Five offspring were pooled to for conducting RT-PCR. Infectious pancreatic necrosis virus (IPNV), infectious hematopoietic necrosis virus (IHNV) and viral hemorrhagic septicemia virus (VHSV) were the target viruses for monitoring. In 2006, only spawners were investigated, and 27.5% of fish (22/80) were found to be IHNV-positive by nested PCR. In 2007, 65.6% of pooled fry (21/32) were IHNV-positive, and 9.4% (3/32) were IPNV-positive by one-step PCR. When nested PCR was conducted, 84.4% (27/32) were IHNV-positive, and 28.1% (9/32) were IPNV-positive. However, only 1.3% of spawners (1/80) were IHNV-positive by nested PCR. In 2008, 25% (8/32) of pooled fry were IHNV-positive by one-step PCR, but 59.4% (19/32) were IHNV-positive and 12.5% (4/32) were IPNV-positive by nested PCR. All of the samples tested were VHSV-negative. Although all viruses detected in this study were from chum salmon, phylogenetic analysis showed that they possibly originated from rainbow trout or clustered with the rainbow trout isolates. More extensive long-term studies are needed to clarify the origins of these viruses and their potential effects on chum salmon migration in Korea.

Atypical Aeromonas salmonicida infection in the black rockfish, Sebastes schlegeli Hilgendorf, in Korea.

Cultured black rockfish, Sebastes schlegeli, suffered mass mortalities during winter 2008 and spring 2009 in Korea, showing clinical signs of ulcer lesions and haemorrhages over their body surface. The aetiological agent was identified as Aeromonas salmonicida (strains RFAS-1, -2 and -3), which is a non-pigmented, slow-growing bacterium. Phenotypes of RFAS strains showed variation, while 16S rRNA, gyrB, rpoD, dnaJ and recA gene sequences of all the strains were affiliated to A. salmonicida. In particular, vapA gene sequences of the strains were most closely related to one of the five subspecies of A. salmonicida subsp. masoucida (=KCCM 40239(T) ). LD(50) values of RFAS-1 for intraperitoneal and intramuscular injection were 1.5 × 10(5.25) and 1.5 × 10(6.4) cfu/rockfish, respectively. However, A. salmonicida strains KCCM 40239(T) and SAS-1, which originate from masou and chum salmon, respectively, were not pathogenic to black rockfish. RFAS strains, possessing A-layer protein on their surface, exhibited ß-haemolytic activity against rockfish erythrocytes and capability to survive in rockfish serum, which seem to be associated with virulence.

Effects of sevoflurane on collagen production and growth factor expression in rats with an excision wound.

BACKGROUND: Sevoflurane is a widely used inhalation anesthetic, but there are no studies on its effect on the wound-healing process. This study was undertaken to evaluate the effect of exposure time to sevoflurane on wound healing. METHOD: Male Sprague-Dawley rats were used. Two circular full-thickness skin defects 8 mm in diameter were made on the dorsum of the rats. The animals were divided into six groups according to exposed gas type and time: S1 (sevoflurane, 1 h), S4 (sevoflurane, 4 h), S8 (sevoflurane, 8 h), O1 (oxygen, 1 h), O4 (oxygen, 4 h), and O8 (oxygen, 8 h). The surface area of the wounds was measured 0, 1, 3, and 7 days after surgery. Separately, the mean blood pressures (MBP) and arterial oxygen pressures (PaO(2)) were monitored during the sevoflurane exposure. Collagen type I production and transforming growth factor-beta1 (TGF-beta1) and basic fibroblast growth factor (bFGF) expression on the wound surface were analyzed. Routine histological analysis was also performed. RESULT: Exposure duration to sevoflurane had no influence on MBP and PaO(2). The reduction in wound size and collagen type I production was delayed in S8. The expression of TGF-beta1 and bFGF on the wound surface in S8 was significantly attenuated in S8. The histology of the S8 demonstrated a delayed healing status. CONCLUSIONS: Prolonged exposure to sevoflurane might alter the inflammatory phase of the wound-healing process by attenuation of growth factor expression such as TGF-beta1 and bFGF and subsequently by reduced collagen production.

Collision metastasis of squamous carcinoma of the oral tongue and incidental thyroid papillary carcinoma to a single cervical lymph node.

The incidence of collision tumor is exceedingly rare. There are only four published case reports. This is the first report of a case of collision metastasis of squamous cell carcinoma (SCC) of the oral tongue and incidental thyroid papillary carcinoma to the same cervical lymph node. A 47-year-old man with SCC of the oral tongue at clinical stage T4N1M0 was treated with total glossectomy and bilateral neck dissection. During neck dissection, concomitant secondary foci of thyroid papillary carcinoma were identified in the same cervical lymph node as SCC (collisional metastasis). The patient subsequently underwent total thyroidectomy and was alive without any recurrences at 25 months after the operation.

Genotyping of Korean isolates of infectious hematopoietic necrosis virus (IHNV) based on the glycoprotein gene.

Glycoprotein (G) gene nucleotide sequences of four Korean isolates of infectious hematopoietic necrosis virus (IHNV) were analyzed to evaluate their genetic relatedness to worldwide isolates. All Korean isolates were closely related to Japanese isolates of genogroup JRt rather than to those of North American and European genogroups. It is believed that Korean IHNV has been most likely introduced from Japan to Korea by the movement of contaminated fish eggs. Among the Korean isolates, phylogenetically distinct virus types were obtained from sites north and south of a large mountain range, suggesting the possibility of more than one introduction of virus from Japan.