Genetic evolution of classical swine fever virus under immune environments conditioned by genotype 1-based modified live virus vaccine.

Modified live vaccines (MLVs) based on genotype 1 strains, particularly C-strain, have been used to prevent and control classical swine fever virus (CSFV) worldwide. Nevertheless, a shift in the predominant CSFV strains circulating in the field from genotype 1 or 3 to genotype 2 is seen. Genotype 2 is genetically distant from the vaccine strains and was recently reported during outbreaks after vaccine failure; this has raised concerns that vaccination has influenced viral evolution. In Korea in 2016, there was an unexpected CSF outbreak in a MLV-vaccinated commercial pig herd. The causative CSFV strain was genetically distinct from previously isolated Korean strains but similar to recent Chinese strains exhibiting enhanced capacity to escape neutralization; this suggests the need for global cooperative research on the evolution of CSFV. We analysed global E2 sequences, using bioinformatics tools, revealing the evolutionary pathways of CSFV. Classical swine fever virus genotypes 1 and 2 experienced different degrees and patterns of evolutionary growth. Whereas genotype 1 stayed relatively conserved over time, the genetic diversity of genotype 2 has progressively expanded, with few fluctuations. It was determined that genotype 2 evolved under lower immune pressures and at a higher evolutionary rate than genotype 1. Further, several selected codons, under diversifying selection in genotype 1 but under purifying selection in genotype 2, correspond to antigenic determinants, which could lead to evasion of vaccine-induced immunity. Our findings provide evidence that evolutionary changes in CSFV are the result of the disproportionate usage of the CSF MLVs in endemic areas; this underscores the need to develop mitigation strategies to minimize the substantial risk associated with the emergence of vaccine-escaping mutants.

The feasibility and efficacy of laparoscopic repair for chronic traumatic diaphragmatic herniation: introduction of a novel technique with literature review.

PURPOSE: Traumatic diaphragm rupture is a rare trauma that is easily overlooked. A missed diagnosis would result in chronic traumatic diaphragmatic herniation (CTDH). Surgical repair is the standard treatment that is conventionally performed by laparotomy or thoracotomy. Laparoscopic repair has been reported, but its efficacy remains controversial. In this study, we present our novel technique and experience of laparoscopic repair of CTDH and analyze the feasibility and effectiveness of this procedure. METHODS: We conducted a prospective collection with retrospective review of patients with CTDH treated at Chang Gung Memorial Hospital, Taiwan, from 2000 to 2013. The demographic characteristics, surgical procedure, perioperative results, length of hospital stay (HLOS) and follow-up were record and analyzed. RESULTS: There were 114 patients with traumatic diaphragm hernia, and 24 of them had CTDH with a mean age of 54.9 ± 13.3 years. The HLOS was 15.08 ± 8.17 days. Regarding the surgical method used, 19 patients had open surgery, and 5 patients underwent laparoscopic surgery. The demographic distribution, trauma mechanism, location and size of CTDH were comparable. In the laparoscopic group, the patients had a shorter median HLOS (6 days) than in the open surgery group (16 days; p = 0.002). There was no mortality or recurrence in both groups. CONCLUSIONS: In this study and literature review, patients had laparoscopic repair with a smooth recovery. Laparoscopy provides good surgical exposure, allowing easy repositioning of the herniated content and a smooth repair of the defect without the morbidity of laparotomy. For CTDH, with caution, we can apply this technique with an acceptable result.

A diterpenoid taxodone from Metasequoia glyptostroboides with antimycotic potential against clinical isolates of Candida species.

OBJECTIVE: The increasing importance of clinical isolates of Candida species and emerging resistance of Candida species to current synthetic antifungal agents have stimulated the search for safer and more effective alternative drugs from natural sources. This study was directed towards exploring the antimycotic potential of a diterpenoid compound taxodone isolated from Metasequoia glyptostroboides against pathogenic isolates of Candida species. MATERIALS AND METHODS: Antimycotic efficacy of taxodone was evaluated by disc diffusion assay, determination of minimum inhibitory (MIC) and minimum fungicidal (MFC) concentrations, and cell viability assay. To confirm a partial antimycotic mode of action of taxodone, the efficacy of taxodone was determined by measuring the release of 260 nm absorbing materials from the selected Candida species as compared to control. RESULTS: The taxodone at the concentration of 400 µg/disc displayed potential antimycotic effect against the tested clinical and pathogenic isolates of Candida species as diameters of zones of inhibitions, which were found in the range of 11 ± 0.0 to 12.6 ± 0.5mm. The MIC and MFC values of taxodone against the tested clinical isolates were found in the range of 250 to 1000 and 500 to 2000µ g/mL, respectively. On the other hand, the MIC and MFC values of positive control (amphotericin B) against the tested Candida isolates were found in the range of 62.5 to 250 and 500 to 2000 µg/mL. On the viable counts of the tested fungal isolates, the taxodone exerted significant antimycotic effect. Elaborative study of partial mode of action conducted onto the release of 260nm materials (DNA and RNA) revealed potential detrimental effect of taxodone on the membrane integrity of the tested pathogens at MIC concentration. CONCLUSION: With respect to the antimycotic effect of taxodone against pathogenic and clinical isolates of Candida species, it might be confirmed that bioactive compound taxodone present in M. glyptostroboides holds therapeutic value of medicinal significance.

In vivo and in vitro ketamine exposure exhibits a dose-dependent induction of activity-dependent neuroprotective protein in rat neurons.

Anesthetic doses of ketamine induce apoptosis, as well as gene expression of activity-dependent neuroprotective protein (ADNP), a putative homeodomain transcription factor in rat pups (P7). This study investigated if ketamine induced ADNP protein in a dose-dependent manner in vitro and in vivo using primary cultures of cortical neurons and neonatal pups (P7). In vivo immunohistochemistry demonstrated a sub-anesthetic dose of ketamine increased ADNP in the somatosensory cortex (SCC) which was previously identified to be damaged by repeated exposure to anesthetic doses of ketamine. Administration of low-dose ketamine prior to full sedation prevented caspase-3 activation in the hippocampus and SCC. Primary cultures of cortical neurons treated with ketamine (10 µM-10mM) at 3 days-in vitro (3 DIV) displayed a concentration-dependent decrease in expanded growth cones. Furthermore, neuronal production and localization of ADNP varied as a function of both ketamine concentration and length of exposure. Taken together, these data support the model that ADNP induction may be partially responsible for the efficacy of a low-dose ketamine pre-treatment in preventing ketamine-induced neuronal cell death.

Risk factors for liver abscess formation in patients with blunt hepatic injury after non-operative management.

PURPOSE: To identify risk factors for liver abscess formation in patients with blunt hepatic injury who underwent non-operative management (NOM). METHODS: From January 2004 to October 2008, retrospective data were collected from a single level I trauma center. Clinical data, hospital course, and outcome were all extracted from patient medical records for further analysis. RESULTS: A total of 358 patients were enrolled for analysis. There were 13 patients with liver abscess after blunt hepatic injury. Patients with abscess had a significant increase in glutamic oxaloacetic transaminase (GOT, p = 0.006) and glutamic pyruvic transaminase (GPT, p < 0.0001), and a decrease in arterial blood pH (p = 0.023) compared to patients without abscess in the univariate analyses. In addition, high-grade hepatic injury and transarterial embolization (TAE, p < 0.001) were also risk factors for liver abscess formation. Five factors (GOT, GPT, pH level in the arterial blood sample, TAE, and high-grade hepatic injury) were included in the multivariate analysis. TAE, high-grade hepatic injury, and GPT level were statistically significant. The odds ratios of TAE and high-grade hepatic injury were 15.41 and 16.08, respectively. A receiver operating characteristic (ROC) analysis was used for GPT, and it suggested cutoff values of 372.5 U/L. A prediction model based on the ROC analysis had 100 % sensitivity and 86.7 % specificity to predict liver abscess formation in patients with two of the three independent risk factors. CONCLUSIONS: TAE, high-grade hepatic injury, and a high GPT level are independent risk factors for liver abscess formation.

Microbial fermentation of cabbage by a bacterial strain of Pectobacterium atrosepticum for the production of bioactive material against Candida species.

OBJECTIVE: The objective of this study was to produce the bioactive fermented product by the microbial fermentation of cabbage (Brassica oleracea) using a bacterial strain Pectobacterium atrosepticum which was assessed for its antimycotic efficacy against pathogenic isolates of Candida species. MATERIALS AND METHODS: An approach of microbial fermentation of cabbage using a bacterial strain P. atrosepticum was applied to obtain the bioactive fermented product. Antimycotic efficacy of bioactive fermented product of cabbage was evaluated by disc diffusion assay, minimum inhibitory (MIC) and minimum fungicidal (MFC) concentrations, cell viability assay and scanning electron microscopy (SEM) analysis. RESULTS: The bioactive fermented product (500µg/disc) revealed promising antimycotic effect against the tested Candida species as a diameter of inhibition zones (10±0.2 to 13±0.4mm) along with its MIC and MFC values, ranging from 250 to 1000 and 250 to 2000µg/ml, respectively. Exposure of 140min of bioactive fermented product exerted potential antimycotic effect on the viable counts of the tested fungal isolates with about 85 to 100% inhibitory effect. Further, the study of SEM revealed potential detrimental effect of bioactive product on the morphology of C. albicans KACC 30003 at MIC concentration. Elaborative study of GC-MS analysis conducted on bioactive fermented product of cabbage revealed transformation products present in fermented product. CONCLUSION: These results confirmed the therapeutic potential of microbially bioconverted/fermented products of cabbage for using in medicinal and pharmaceutical preparations.

Studies on the viability and membrane integrity of human spermatozoa treated with essential oil of Trachyspermum ammi (L.) Sprague ex Turrill fruit.

The present study aimed at investigating the effects of essential oil of Trachyspermum ammi fruits, an oil-bearing plant of Apiaceae family, on human sperm viability and membrane integrity. Chemical compositions of the oil were analysed by GC-MS. Thirty compounds representing 91.39% of the total oil were identified. The viability and membrane integrity of human spermatozoa were assessed using minimum effective dose (MED) concentration (125 µg ml(-1)) of the oil. Sperm treated with essential oil showed a significant decrease (P < 0.05) in viability assessed by eosin-nigrosin and fluorescence dual staining. Moreover, the treated sperm also showed a significant loss (P < 0.05) of functional mitochondria and antioxidant enzyme, catalase (EC 1.11.1.6, CAT), when compared to control. The cholesterol:phospholipid ratio was also increased (P < 0.05) in treated sperm when compared to control, which is an indicator of loss of binding ability of human spermatozoa to the zona pellucida. The scanning electron microscopic studies demonstrated the loss of membrane integrity in essential oil-treated human spermatozoa, which showed vacuolation, swelling of acrosomal cap, detachment of head portion and tail coiling. Present observations indicate the spermicidal property of essential oil of T. ammi fruits, which could be helpful to develop medicinal preparations as a male contraceptive.

Adult liver abscess: characteristics of patients living in rural north-eastern Taiwan.

AIMS: This study aimed to analyse the characteristics of adult liver abscess (LA) patients living in rural townships of Taiwan. PATIENTS AND METHODS: We retrospectively screened the electronic admission records of a rural community hospital located in north-eastern Taiwan from 1 April, 2002 to 30 April, 2006. Relevant data, including subjects' basic characteristics, laboratory findings and infectious microorganisms, were extracted. RESULTS: Fifty-six subjects (mean age: 66.1 +/- 15.9 years; range: 23-94 years) were enrolled; one patient had an amoebic LA and 55 had pyogenic LA. Five subjects died in hospital. Overall, 80.5% of patients complained of having a fever, 87.5% had single abscess, 71.4% had right hepatic lobe involvement and 58.9% underwent invasive drainage. Most subjects (66.1%) did not have diabetes mellitus, 94.6% did not have a hepato-biliary tumour, 73.2% did not have gallstones, 78.6% did not have hepatitis and 87.5% did not have prior hepato-biliary surgery. Klebsiella pneumoniae was the most common infecting microorganism. Previous hepato-biliary surgery and serum creatinine >2.0 mg/dl were significantly more common in patients >or=65 years of age (p = 0.031). Diabetes mellitus was more common in female subjects (p = 0.021). Invasive drainage and single abscess were not significantly correlated to prognosis. DISCUSSION: Adult LA patients living in rural north-eastern Taiwan have different characteristics than patients living in urban areas. Geriatric LA patients should be managed cautiously because of the possibility of renal insufficiency or previous hepato-biliary surgery. Female LA patients should be evaluated for the presence of diabetes mellitus.

Process of rock phosphate solubilization by Aspergillus sp PS 104 in soil amended medium.

Aspergillus sp PS 104, a soil isolate had excellent potential to solubilize rock phosphate in vitro. The process was influenced by the presence of various concentrations of local loess (red soil). The simultaneous occurrence, in our experiment, of high levels of solubilized phosphate and synthesized citric acid, together with the lowest reached pH values, confirmed the role of citric acid in the phosphate solubilization mechanism. When the soil was present, phosphate release was better correlated than citrate synthesis with H+ concentration. Changes in soluble phosphate concentration did not follow a sigmoid pattern. The ability of organism to release phosphatase was also studied. An interesting relationship was observed between the two processes of phosphate mobilization: citric acid synthesis and phosphatase production.

In vitro inhibition of food spoilage and foodborne pathogenic bacteria by essential oil and leaf extracts of Magnolia liliflora Desr.

The aim of this study was to examine the chemical composition of the essential oil isolated from the floral parts of Magnolia liliflora Desr. by hydrodistillation, and to test the efficacy of essential oil and various leaf extracts against a diverse range of microorganisms comprising food spoilage and foodborne pathogenic bacteria. The chemical composition of essential oil was analyzed by GC-MS. It was determined that 52 compounds, which represented 78.07% of total oil, were present in the oil. The oil contained mainly levoxine (15.59%), methylcyclopropane (24.26%), 2-beta-pinene (5.3%), caryophyllene oxide (4.01%), and beta-caryophyllene (1.7%). The oil (1000 ppm/disc) and leaf extracts (1500 ppm/disc) exhibited promising antibacterial effects against the tested pathogens as a diameter of zones of inhibition (9 to 18 and 7 to 12 mm) and MIC values (125 to 1000 and 500 to 3000 microg/mL), respectively. Also, the oil had a potent detrimental effect on the viable count of the tested bacteria. The results obtained in this study support the role of essential oil and the leaf extracts derived from M. liliflora as a remarkable approach to inhibit the growth of food spoilage and foodborne pathogens.

Association between estrogen receptor polymorphism and pain susceptibility in female temporomandibular joint osteoarthritis patients.

The purpose of this study was to investigate the possible association between estrogen receptor alpha (ERalpha) polymorphism and pain susceptibility in female symptomatic temporomandibular joint (TMJ) osteoarthritis (OA) patients. A patient group of 100 women, diagnosed as TMJ OA according to the research diagnostic criteria for temporomandibular disorders, were selected, and 74 women with no signs and symptoms of temporomandibular disorder were assigned to a control group. Pvu II and Xba I restriction fragment length polymorphisms were analyzed by direct haplotyping. The patient group was divided into three subgroups according to a visual analog scale (VAS): mild pain (0.05). TMJ OA patients carrying the PX haplotype were found to have a significantly higher risk of moderate or severe pain compared to those without the PX haplotype, suggesting that ERalpha polymorphism may be associated with pain susceptibility in female TMJ OA patients.

Pyoderma gangrenosum.

Pyoderma Gangrenosum (PG) is a non-infective, necrotising neutrophilic dermatosis. Many diseases may mimic PG. It may be idiopathic or may be associated with a systemic disorder. We report a case of PG in association with ulcerative colitis.

Distribution of persistent organic pollutants in soil and grasses around Mt. Qomolangma, China.

Previous literature has reported the fate of persistent organic pollutants (POPs) in mountainous regions, but the Himalayas have received little attention, and few results from this region have been published. The present study collected soil and grass samples from the Mt. Qomolangma (Everest) area, central Himalayas, China, from the elevation range 4700 to 5620 m. We analyzed all samples for organochlorine pesticides (OCPs) to determine the level of OCP contamination in the Qomolangma region. The soil samples contained 0.385 to 6.06 ng g(-1) of DDT only, and these concentrations were lower than those from Europe and mountains close to industrial emissions. Our study detected a number of OCPs in the grass samples, such as hexachlorocyclohexane (HCH) (0.354 to 7.82 ng g(-1)), hexachlorobenzene (HCB) (0.0156 to 1.25 ng g(-1)), endosulfan (0.105 to 3.14 ng g(-1)), and DDT components (1.08 to 6.99 ng g(-1)). Their concentrations were higher than those in pine needles from Alberta, Canada. Our measurements of HCH and DDT in grass samples showed the same or slightly higher concentration levels than reported in moss from Mt. Qomolangma 15 years ago. This result and the analysis of isomer ratios (alpha/gamma-HCH and p-p'-DDE/p-p'-DDT) indicate recent releases of OCPs from a nearby region, possibly from dicofol use in India. We also investigated the elevation distribution of OCPs and found that HCH and HCB were progressively concentrated in colder, higher elevation sites. A bioconcentration factor (BCF) of grass was calculated, and the BCF values increased with the increasing elevation, indicating that the cold condensation of POPs at high-elevation sites may increase the potential threat to vegetation and the food chain in the mountain ecosystem.

Two years review of cutaneous adverse drug reaction from first line anti-tuberculous drugs.

First line Anti-TB therapy with rifampicin, isoniazid, pyrazinamide, and ethambutol/streptomycin is very effective. However, major adverse reactions to antituberculous drugs can cause significant morbidity and mortality. Cutaneous adverse drug reaction (CADR) is one of the commonly observed major adverse events. This retrospective study looked at the cases of TB treated in Respiratory Unit, Penang Hospital from January 2004 to December 2005. Of 820 patients treated for active TB, 47 patients (25 females; 22 males) developed CADR (5.7%). CADRs observed include morbiliform rash (72.3%), erythema multiforme syndrome (8.5%), urticaria (8.5%) and others (which include exfoliative dermatitis and lichenoid eruption). Ninety-seven percent of events occurred within two months after the initial dose. Incidence rate of CADR among the first line anti-TB drugs, pyrazinamide was the commonest offending drug (2.38%), followed by streptomycin (1.45%), ethambutol (1.44%), rifampicin (1.23%) and isoniazid (0.98%). Various clinical characteristics of patients with CADR identified include Human Immunodeficiency Virus (HIV) infection (27.7%), polypharmacy (21.3%), elderly (19.1%), autoimmune disorders (6.4%), pre-existing renal impairment (4.3%), pre-existing liver disorders (4.3%). In conclusion, CADR is common and majority of cases occurred within two months after initiation of anti-TB treatment, particularly in HIV infected patients. Pyrazinamide is the commonest offending drug.

Estrogen receptor gene polymorphism and craniofacial morphology in female TMJ osteoarthritis patients.

The aim of this study was to investigate the possible influence of estrogen receptor alpha (ERalpha) polymorphism on the craniofacial skeleton in female patients suffering from symptomatic osteoarthritis (OA) of the temporomandibular joint (TMJ). The sample comprised 76 genetically unrelated Korean women diagnosed with OA by research diagnostic criteria for temporomandibular disorders (RDC-TMD). Direct haplotyping procedure was used to analyze the PvuII and XbaI RFLPs. Twelve cephalometric measurements were taken to evaluate the spatial position and dimensions of the mandible. Mann-Whitney's U-test was used to identify the potential differences in the cephalometric measurements between the subjects grouped according to their carrier status for Px haplotype. In addition, an association study was carried out using chi(2)-test to further examine the relationship between Px haplotype and the craniofacial morphology of the symptomatic OA patients. Female symptomatic TMJ OA patients carrying Px haplotype showed significantly smaller facial axis angle (P<0.05) and mandibular body length (P<0.05) than the non-carriers. The association between the presence of Px haplotype and short mandibular body length was also ascertained. This study suggests that ERa polymorphism contributes to the altered mandibular dimensions in female symptomatic TMJ OA patients. Further studies on the role of the genetic markers relevant to the craniofacial growth and adaptation are expected to broaden our understanding of determinants of the craniofacial morphology.

Mapping the antigenicity of copper-treated cellular prion protein with the scrapie isoform.

When recombinant and cellular prion protein (PrP(C)) binds copper, it acquires properties resembling the scrapie isoform (PrP(Sc)), namely protease resistance, detergent insolubility and increased beta sheet content. However, whether the conformations of PrP(C) induced by copper and PrP(Sc) are similar has not been studied in great detail. Here, we use a panel of seven monoclonal antibodies to decipher the epitopes on full-length mouse PrP(C) that are affected by exogenous copper, and to compare the antigenicity of the copper-treated full-length PrP(C) with the full-length PrP(Sc) present in scrapie-infected mouse brains. In the presence of copper, we found that epitopes along residues 115-130 and 153-165 become more accessible on PrP(C). These regions correspond to the two beta sheet strands in recombinant PrP and they were proposed to be important for prion conversion. However, when we compared the antibody-binding patterns between full-length PrP(C) with full-length PrP(Sc) and between copper-treated full-length PrP(C) with full-length PrP(Sc), antibody binding to residues 143-155 and 175-185 was consistently increased on PrP(Sc). Collectively, our results suggest that copper-treated full-length PrP(C) does not resemble full-length PrP(Sc), despite acquiring PrP(Sc)-like properties. In addition, since each full-length protein reacts distinctively to some of the antibodies, this binding pattern could discriminate between PrP(C) and PrP(Sc).

Rhizobia as a biological control agent against soil borne plant pathogenic fungi.

Rhizobia promote the growth of plants either directly through N2 fixation, supply of nutrients, synthesis of phytohormones and solubilization of minerals, or indirectly as a biocontrol agent by inhibiting the growth of pathogens. The biocontrol effect of rhizobia is due to the secretion of secondary metabolites such as antibiotics and HCN. Siderophore production in iron stress conditions provides rhizobia an added advantage, resulting in exclusion of pathogens due to iron starvation.

Experimental and theoretical analysis of wavelet-based denoising filter for echocardiographic images.

One of the most significant features of diagnostic echocardiographic images is to reduce speckle noise and make better image quality. In this paper we proposed a simple and effective filter design for image denoising and contrast enhancement based on multiscale wavelet denoising method. Wavelet threshold algorithms replace wavelet coefficients with small magnitude by zero and keep or shrink the other coefficients. This is basically a local procedure, since wavelet coefficients characterize the local regularity of a function. After we estimate distribution of noise within echocardiographic image, then apply to fitness Wavelet threshold algorithm. A common way of the estimating the speckle noise level in coherent imaging is to calculate the mean-to-standard-deviation ratio of the pixel intensity, often termed the Equivalent Number of Looks(ENL), over a uniform image area. Unfortunately, we found this measure not very robust mainly because of the difficulty to identify a uniform area in a real image. For this reason, we will only use here the S/MSE ratio and which corresponds to the standard SNR in case of additivie noise. We have simulated some echocardiographic images by specialized hardware for real-time application;processing of a 512*512 images takes about 1 min. Our experiments show that the optimal threshold level depends on the spectral content of the image. High spectral content tends to over-estimate the noise standard deviation estimation performed at the finest level of the DWT. As a result, a lower threshold parameter is required to get the optimal S/MSE. The standard WCS theory predicts a threshold that depends on the number of signal samples only.

Use of nonsingular boundary integral formulation for reducing errors due to near-field measurements in the boundary element method based near-field acoustic holography.

In the conformal near-field acoustic holography (NAH) using the boundary element method (BEM), the transfer matrix relating the vibro-acoustic properties of source and field depends solely on the geometrical condition of the problem. This kind of NAH is known to be very powerful in dealing with the sources having irregular shaped boundaries. When the vibro-acoustic source field is reconstructed by using this conformal NAH, one tends to position the sensors as close as possible to the source surface in order to get rich information on the nonpropagating wave components. The conventional acoustic BEM based on the Kirchhoff-Helmholtz integral equation has the singularity problem in the close near field of the source surface. This problem stems from the singular kernel of the Green function of the boundary integral equation (BIE) and the singularity can influence the reconstruction accuracy greatly. In this paper, the nonsingular BIE is introduced to the NAH calculation and the holographic BIE is reformulated. The effectiveness of nonsingular BEM has been investigated for the reduction of reconstruction error. Through interior and exterior examples, it is shown that the resolution of predicted field pressure could be improved in the close near field by employing the nonsingular BIE. Because the BEM-based NAH inevitably requires the field pressure measured in the close proximity to the source surface, the present approach is recommended for improving the resolution of the reconstructed source field.