Effect of additive-assisted fat transplantation on fat graft survival rate: A preliminary experimental study based on a rabbit animal model.

BACKGROUND: Improving the survival rate of fat grafts is yet a difficult problem in the field of autologous fat transplantation. Prevailing methods such as making nanofat and SVF are time-consuming. Hence, the role of additives application in the improvement of fat graft survival during fat transplantation was considered and preliminarily evaluated in a rabbit animal model. METHODS: A rabbit animal model was established where rabbit ears were injected with a mixture of 1.5mL of adipose tissue and 1mL of saline (group A), 1.5mL of adipose tissue and 1mL of botulinum toxin A (BoNTA) (group B), 1.5mL of adipose tissue and 1mL of prostaglandin E2 (groupC), 1.5mL of adipose tissue and 1mL of PDRN (group D) respectively. Then, the extents of neovascularization and inflammation were evaluated on the 7th, 14th, 28th, 42nd, 56th and 70th day after injection by ELISA assays and H&E and immunofluorescence staining. RESULTS: The results showed that pre-treatment with BoNTA, prostaglandin E2 and PDRN improved graft volume and weight. The H&E and immunofluorescence staining revealed that BoNTA, prostaglandin E2 and PDRN improved the graft angiogenesis. Simultaneously, TNF-α expression level detected by ELISA was the lowest in the PDRN group. CONCLUSION: Henceforth, the present preliminary study suggests that pre-transplantation treatment with BoNTA, prostaglandin E2 and PDRN can improve the fat graft angiogenesis and graft integrity, whereby the effect of adding PDRN may be significant.

MR elastography of the liver: comparison of three measurement methods.

AIM: To analyse the inter- and intra-observer agreement of liver stiffness value (LSV) using three methods with 3 T magnetic resonance elastography (MRE) and to investigate factors related to LSV difference. MATERIALS AND METHODS: This retrospective study included 147 patients. Two independent observers measured the LSV using three region of interest (ROI) methods: (1) circular ROI with a radius of 1 cm in the right lobe, (2) largest ROI possible, and (3) average value considering the measurement area. The agreement and factors related to difference of LSV were investigated. RESULTS: The intraclass correlation coefficients were excellent at 0.982-0.997 in all methods. The differences between observers for method 1 were significantly larger than those of method 2 or method 3 (p<0.001). The Child-Pugh classification was only related to LSV difference (p<0.001). CONCLUSION: Methods 2 and 3 were significantly more reliable than method 1. The Child-Pugh classification was only related to difference of LSV.

H3N2 canine influenza virus with the matrix gene from the pandemic A/H1N1 virus: infection dynamics in dogs and ferrets.

After an outbreak of pandemic influenza A/H1N1 (pH1N1) virus, we had previously reported the emergence of a recombinant canine influenza virus (CIV) between the pH1N1 virus and the classic H3N2 CIV. Our ongoing routine surveillance isolated another reassortant H3N2 CIV carrying the matrix gene of the pH1N1 virus from 2012. The infection dynamics of this H3N2 CIV variant (CIV/H3N2mv) were investigated in dogs and ferrets via experimental infection and transmission. The CIV/H3N2mv-infected dogs and ferrets produced typical symptoms of respiratory disease, virus shedding, seroconversion, and direct-contact transmissions. Although indirect exposure was not presented for ferrets, CIV/H3N2mv presented higher viral replication in MDCK cells and more efficient transmission was observed in ferrets compared to classic CIV H3N2. This study demonstrates the effect of reassortment of the M gene of pH1N1 in CIV H3N2.

Association between an HLA-G 14 bp insertion/deletion polymorphism and non-segmental vitiligo in the Korean population.

Vitiligo is a pigmentary skin disorder characterized by the chronic and progressive loss of melanocytes. Although the etiology of vitiligo is still unknown, several theories have been proposed to explain the pathogenesis of vitiligo including autoimmune, neural, self-destruction, oxidative stress, and genetic theories. Human leukocyte antigen (HLA)-G is a nonclassic, major histocompatibility complex class I molecule that plays an important role in suppression of the immune response. Several recent studies have provided evidence that a 14 bp insertion (INS)/deletion (DEL) polymorphism in the HLA-G gene might be associated with autoimmune disease. Our aim in this study was to determine whether the 14 bp INS/DEL polymorphism in the HLA-G gene contributes to the risk of developing non-segmental vitiligo (NSV) in the Korean population. We conducted a case-control association study of 192 NSV patients and 491 matched, unaffected controls. The HLA-G 14bp INS/DEL polymorphism was analyzed by gene scan after amplification using the polymerase chain reaction. Genotype frequencies for the 14bpINS/DEL were different between the vitiligo group and Korean control group. The proportion of subjects with a homozygote 14bpINS/14bpINS genotype was significantly higher in the vitiligo group compared with the control group (7.1 vs. 3.5 %, OR 2.25, 95 % CI 1.06-4.76, p = 0.039 in the recessive model). Our results suggest that the HLA-G 14bpINS/DEL polymorphism is associated with the development of NSV in the Korean population.

Prevalence and different characteristics of two serotypes of Streptococcus parauberis isolated from the farmed olive flounder, Paralichthys olivaceus (Temminck and Schlegel), in Korea.

The prevalence of two serotypes of Streptococcus parauberis isolated from the olive flounder, Paralichthys olivaceus, was evaluated in a total of 29 isolates between 2003 and 2010 in Korea. Streptococcus parauberis isolates were divided into two serologically distinct types (serotype 1 and serotype 2), except for one strain (S1091), using an agglutination assay with rabbit antiserum, and serotype 1 was identified as the dominant type (24 of 29 isolates) in this study. To identify the characteristics of the two serotypes of S. parauberis, we conducted a biochemical test using the API 20 Strep kit, a transmission electron microscopy (TEM) assay, sequence analysis of 16S-23S rRNA intergenic spacer region (ISR) and a pathogenicity test. In TEM, both serotypes possessed polysaccharide capsule layers around the cell surface when bacterial cells were treated with a homologous serotype of rabbit antiserum. However, we were unable to discriminate serotype-specific biochemical characteristics and genetic characteristics of 16S-23S rRNA ISR between the two serotypes. In the pathogenicity test, the serotype 1 strains induced significantly higher mortality than the serotype 2 strains in olive flounder when experimentally inoculated via the intraperitoneal route.

Interspecies transmission of the canine influenza H3N2 virus to domestic cats in South Korea, 2010.

In the past 4 years, incidences of endemic or epidemic respiratory diseases associated with canine influenza H3N2 virus in Asian dogs have been reported in countries such as South Korea and China. Canine species were considered to be the new natural hosts for this virus. However, at the beginning of 2010, influenza-like respiratory signs, such as dyspnoea, were also observed among cats as well as in dogs in an animal shelter located in Seoul, South Korea. The affected cats showed 100 % morbidity and 40 % mortality. We were able to isolate a virus from a lung specimen of a dead cat, which had suffered from the respiratory disease, in embryonated-chicken eggs. The eight viral genes isolated were almost identical to those of the canine influenza H3N2 virus, suggesting interspecies transmission of canine influenza H3N2 virus to the cat. Moreover, three domestic cats infected with intranasal canine/Korea/GCVP01/07 (H3N2) all showed elevated rectal temperatures, nasal virus shedding and severe pulmonary lesions, such as suppurative bronchopneumonia. Our study shows, for the first time, that cats are susceptible to canine influenza H3N2 infection, suggesting that cats may play an intermediate host role in transmitting the H3N2 virus among feline and canine species, which could lead to the endemic establishment of the virus in companion animals. Such a scenario raises a public health concern, as the possibility of the emergence of new recombinant feline or canine influenza viruses in companion animals with the potential to act as a zoonotic infection cannot be excluded.

Persistence of propargite on okra under subtropical conditions at Ludhiana, Punjab, India.

Residues of propargite were estimated in okra fruits by high performance liquid chromatography following single application of Omite 57 EC @570 and 1,140 g a.i./ha. Residues of propargite were confirmed by high performance thin layer chromatography. The average initial deposits of propargite were observed to be 1.36 and 3.32 mg/kg, respectively, which were below its maximum residue limit of 2 mg/kg. The residue levels of propargite dissipated below its limit of quantification of 0.02 mg/kg after 7 days at single dose and after 10 days at double dose. The half-life values (T 1(/)2) of propargite were worked out to be 0.79 and 0.73 days, respectively, at recommended and double the recommended dosages.

Pathology in dogs with experimental canine H3N2 influenza virus infection.

Avian-lineage H3N2 canine influenza virus (CIV)-associated respiratory disease, which can be fatal, emerged in South Korean dogs in 2007. We show here that dogs experimentally infected with CIV only developed respiratory tract diseases, as no extrapulmonary lesions and virus antigens were detected. This differs from the multiorgan diseases that avian influenza H5N1 induces in small experimental animals. However, the CIV-infected dogs developed a distinctively severe, long-persistent bronchointerstitial pneumonia, which differs from the acute but short-term bronchopneumonia that human (H1N1 and H3N2) influenza cause in rodents and ferrets. Histopathology and in situ TUNEL assays revealed that the neutrophils infiltrating the lesions were undergoing apoptosis, which probably reflects the attempts by the body to maintain appropriate numbers of neutrophils for defense against secondary bacterial infections. Our observations suggest that neutrophils along with the related chemoattractant cytokines (TNF-alpha, IL-1 and IL-8, etc.) may play a key role in the pathogenesis of H3N2 CIV in dogs.

Dissipation kinetics of propargite in brinjal fruits under subtropical conditions of Punjab, India.

Residues of propargite were estimated in brinjal fruits by High Performance Liquid Chromatography (HPLC) following single application of Omite 57 EC @ 570 and 1140 g a.i./ha. The average initial deposits of propargite were observed to be 0.51 and 0.92 mg/kg, respectively, which were below its maximum residue limit (MRL) of 2 mg/kg. The residue levels of propargite dissipated below limit of quantification (LOQ) of 0.02 mg/kg after 10 days at both the dosages. The half-life values (T(1/2)) of propargite were worked out to be 3.07 and 3.54 days, respectively, at recommended and double the recommended dosages. A waiting period of one day has been suggested for the safe consumption of brinjal fruits to avoid any health hazards.

Phylogenetic analysis of swine influenza viruses recently isolated in Korea.

Several influenza A viral subtypes were isolated from pigs during a severe outbreak of respiratory disease in Korea during 2005 and 2006. They included a classical swine H1N1 subtype, two swine-human-avian triple-recombinant H1N2 subtypes, and a swine-human-avian triple-recombinant H3N2 subtype. In the current study, genetic characterization to determine the probable origin of these recent isolates was carried out for the first time. Phylogenetic analysis indicated that all the recent Korean isolates of H1N1, H1N2, and H3N2 influenza are closely related to viruses from the United States. Serologic and genetic analysis indicated that the Korean H1N2 viral subtypes were introduced directly from the United States, and did not arise from recombination between Korean H1N1 and H3N2. We suggest that the H1N1, H1N2, and H3N2 viral subtypes that were isolated from the Korean swine population originated in North America, and that these viruses are currently circulating in the Korean swine population.

One-step multiplex RT-PCR for detection and subtyping of swine influenza H1, H3, N1, N2 viruses in clinical samples using a dual priming oligonucleotide (DPO) system.

The swine influenza virus (SIV) H1N1, H1N2, and H3N2 subtypes circulate in Korean farm. A novel multiplex RT-PCR (m-RT-PCR) was developed to detect and subtype swine influenza viruses. This m-RT-PCR assay could identify H1, H3, N1 and N2 from clinical samples in single tube reaction using DPO system. Korean SIVs are closely related to the United States influenza viruses, and primers were developed for SIV from North American viruses and recently Korean isolates. The sensitivity of the m-RT-PCR was 10TCID(50)/ml for H1N1, H1N2 or H3N2. The lowest viral concentrations detected by single PCR were 1TCID(50)/ml for each subtype. Non-specific reactions were not observed when other viruses and bacteria were used to assess the m-RT-PCR. The results of m-RT-PCR were more effective than virus isolation or hemagglutination (HA) test. This assay using a DPO system provides a rapid, sensitive, and cost-effective laboratory diagnosis for detecting and subtyping of SIV in pigs.

Elimination of porcine reproductive and respiratory syndrome virus from a seedstock breeding farm and a supplying boar stud by a modified test and removal method.

To eliminate porcine reproductive and respiratory syndrome virus (PRRSV) from a supplying boar stud, samples of serum and semen from 118 boars were assessed three times a month by an indirect fluorescent antibody (IFA) test to detect antibodies, and by a nested reverse transcriptase-PCR (nRT-PCR) to detect the genome of PRRSV. The boars detected as persistently infected carriers were culled. A PRRSV-negative population of boars was established after three months and no semen positive for the virus was detected for six months. Subsequently, a three-step plan was introduced to eliminate PRRSV from the seedstock breeding farm during three parity cycles on the farm over 15 months, each step taking five months. In step 1, umbilical cords taken from piglets at birth and serum samples taken from their dams at the start of weaning were subjected to ifa and nRT-pcr analysis. The sows with PRRSV detected in serum by nRT-pcr were regarded as carrier sows and culled. The rates of detection of PRRSV were reduced from 5 percent to 2.5 percent in the sera of the sows, and from 14.8 percent to 1.8 percent in the umbilical cords of the piglets. In step 2, the sows that had farrowed the piglets with PRRSV detected by nRT-PCR in their cords were considered to have transmitted the infection and removed. During step 2, the virus detection rates in umbilical cords by nRT-pcr were reduced, but not completely eliminated. In step 3, 10-week-old nursery pigs with antibodies to PRRSV in their serum by ifa and elisa were culled. The three steps established the PRRSV-negative state of the multisite farm containing the breeding and nursery farm, and the PRRSV-negative state of both the multisite farm and the supplying boar stud was evaluated by monthly monitoring over at least one parity cycle of the farm for five months.

Oral efficacy of Vero cell attenuated porcine epidemic diarrhea virus DR13 strain.

A Vero cell attenuated porcine epidemic diarrhea virus (PEDV) strain, DR13, was distinguished from wild-type PEDV using restriction enzyme fragment length polymorphism (RFLP). Cell attenuated DR13 was orally or intramuscularly (IM) administered to late-term pregnant sows, and mortality resulting from the highly virulent PEDV challenge was investigated in passively immunized suckling piglets of the two different groups. The mortality rate of the oral group (13%) was lower than that of the IM group (60%). In particular, the concentration of IgA against PEDV was higher in piglets of sows in the oral group, compared to the IM group. The attenuated DR13 virus remained safe, even after three backpassages in piglets. The findings of this study support the theory that the Vero cell attenuated DR13 virus may be applied as an oral vaccine for inducing specific immunity in late-term pregnant sows with a high margin of protection against PEDV infection.

Isolation and phylogenetic analysis of H1N1 swine influenza virus isolated in Korea.

A swine influenza H1N1 virus was isolated from a pig during a severe outbreak of respiratory disease in Korea. All genes of the H1N1 isolate, including hemagglutinin (HA), neuraminidase (NA), matrix (M), nucleoprotein (NP), non-structural (NS), PA, PB1 and PB2, were of swine origin. Also, all these genes showed a close phylogenic relationship with those of H1N1 viruses previously isolated from pigs in the United States. These results suggest that North American swine influenza virus has actually been transmitted to pigs in Korea.

Risk assessment through dietary intake of total diet contaminated with pesticide residues in Punjab, India, 1999-2002.

Forty-six samples each of vegetarian and nonvegetarian total diet consumed from March 1999 to December 2002 by male subjects in the age group of 19-24 years were analyzed to assess their risk through dietary intake with respect to pesticide residues. The results revealed low dietary intake of levels of Sigma-dichlorodiphenyltrichloroethane (DDT) and which were almost comparable to levels reported in developed countries. The results are indicative of contamination of total diet with pesticide residues despite a ban on the use of DDT and restricted use of lindane in agriculture only. Predominance of lindane residues indicates that liquid milk was a main contributory source as it comprises almost 21% to the total diet consumed per day. Concerted efforts by regulatory authorities and emphasis on judicious use of agrochemicals in pest control are required to decrease the burden of these chemicals in food stuffs to levels safe for dietary intake.

Contamination of liquid milk and butter with pesticide residues in the Ludhiana district of Punjab state, India.

An analysis of 92 samples of liquid milk from Ludhiana, India, during 1999-2001 revealed the presence of DDT in 6 (7.4%) samples and of these 2 samples were found to exceed the maximum residue limit (MRL) of DDT fixed at 0.05 mg kg(-1) (on a whole milk basis). HCH residues were detected in 49 (53.3%) samples and constituted only gamma-HCH (lindane). The MRL of lindane is fixed at 0.01 mg kg(-1) (whole milk basis), and all 49 liquid milk samples exceeded this value. These results are indicative of contamination of bovine milk with pesticide residues as a result of the ban on the use of DDT and HCH in agriculture and public health programs. Similarly, analysis of 40 samples of butter showed the presence of DDT and HCH in 28 and 8 samples, respectively. However, none of the samples exceeded the MRL value of either DDT or any isomer of HCH. DDT residues comprised mainly p,p-DDE and p,p-TDE, whereas HCH residues were present as lindane in 6 samples, and 2 samples revealed the presence of beta-HCH. The estimated daily intake of lindane through the consumption of contaminated liquid milk exceeded its acceptable daily intake value for children. Interestingly, none of the liquid milk or butter samples revealed the presence of any commonly used organophosphorus or synthetic pyrethroid insecticides at their detection limit of 0.01 mg kg(-1).

Characterization of an Aedes aegypti bacterial artificial chromosome (BAC) library and chromosomal assignment of BAC clones for physical mapping quantitative trait loci that influence Plasmodium susceptibility.

Previous studies have confirmed a genetic basis for susceptibility of mosquitoes to Plasmodium parasites. Here we describe our efforts to characterize a bacterial artificial chromosome genomic library for the yellow fever mosquito, Aedes aegypti, and to identify BAC clones containing genetic markers that define quantitative trait loci (QTL) for Plasmodium gallinaceum susceptibility. This library (NDL) was prepared from the Ae. aegypti Liverpool strain and consists of 50 304 clones arrayed in 384-well microplates. We used PCR analysis with oligonucleotide primer pairs specific to 106 genetic markers (as sequence-tagged sites or STS) to screen the NDL library. Each STS identified between one and thirteen independent clones with an average of 3.3 clones. The average insert size was 122 kb and therefore the NDL library provides approximately 7.87-fold genome coverage. The availability of the NDL library should greatly facilitate physical mapping efforts, including positional cloning of QTL for traits of interest such as Plasmodium susceptibility and for whole genome sequence determination and assembly.

Effect of an extract of the root of Scutellaria baicalensis and its flavonoids on aflatoxin B1 oxidizing cytochrome P450 enzymes.

The inhibition of aflatoxin B1 (AFB1) metabolism by a water extract of the root of Scutellaria baicalensis and its flavonoids was examined in liver microsomes. AFB1 is known to be metabolized to aflatoxin M1 (AFM1), aflatoxin Q1 (AFQ1), and AFB1-8,9-epoxide (AFBO). The water extract potently inhibited the production of AFM1 by cytochrome P450 (CYP)1A1/2 and slightly reduced AFBO formation by CYP1A1/2, CYP2B1, CYP2C11 and CYP3A1/2 in TCDD-treated rat liver microsomes. IC50 values for AFM1 and AFBO formation were 6.8 and 122.4 microg/ml, respectively. Wogonin showed the highest inhibitory activity towards AFM1 formation among the flavonoids isolated from the extract. On the other hand, the extract had no effects on the formation of AFBO and AFQ1 in human liver microsomes, and on the activities of CYP2B1, CYP2C11 and CYP3A1/2 which were detected by hydroxylation patterns of testosterone. These results demonstrated that the extract of the root of Scutellaria baicalensis has a specific inhibitory effect on CYP1A1/2 among CYP enzymes involved in AFB1 metabolism by rat and human microsomes.