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The allergenic and toxicological acceptances of the bio-elicited peanut sprout powder (BPSP) have not been assessed. BPSP was generated from peanut kernels germinated at 26-28 °C for 72 h (designated as 72 h-NGS). The 72 h-NGS were subsequently sliced, incubated, dried, defatted and pulverized to generate bio-elicited peanut sprout powder (BPSP). Protein solubility of BPSP increased 2.6-fold compared to 72 h-NGS. SDS-PAGE analysis revealed BPSP production triggered extensive degradation of the high-molecular weight peanut allergic proteins, mainly Ara h 1 and Ara h 3. Western blotting detected with peanut allergic patients' IgE indicated decreased in vitro reactivity. Food safety assessment of BPSP was performed with ICR mice fed with basal (control) and three doses of formulated BPSP-supplemented diets containing 0.11 g (normal), 2.5 g (high) and 25 g (super high) BPSP /kg BW. Animals appeared healthy with steady body weight gain in all groups during the entire 35-day dietary intervention. Hematological and serum biochemical analyses revealed no significant difference among groups. Histopathological examination on the tissue sections of primary organs further supported safety with no pathologies. The in vitro allergic reduction and toxicological safety in the BPSP-supplemented dietary intervention in the ICR mice study, support moving forward with BPSP-involved product development.The online version contains supplementary material available at 10.1007/s13197-022-05537-7.

Development of targeted therapies will be a critical step towards reducing the mortality associated with triple-negative breast cancer (TNBC). To achieve this, we searched for targets that met three criteria: (1) pharmacologically targetable, (2) expressed in TNBC, and (3) expression is prognostic in TNBC patients. Since nuclear receptors have a well-defined ligand-binding domain and are thus highly amenable to small-molecule intervention, we focused on this class of protein. Our analysis identified TLX (NR2E1) as a candidate. Specifically, elevated tumoral TLX expression was associated with prolonged recurrence-free survival and overall survival for breast cancer patients with either estrogen receptor alpha (ERα)-negative or basal-like tumors. Using two TNBC cell lines, we found that stable overexpression of TLX impairs in vitro proliferation. RNA-Seq analysis revealed that TLX reduced the expression of genes implicated in epithelial-mesenchymal transition (EMT), a cellular program known to drive metastatic progression. Indeed, TLX overexpression significantly decreased cell migration and invasion, and robustly decreased the metastatic capacity of TNBC cells in murine models. We identify SERPINB2 as a likely mediator of these effects. Taken together, our work indicates that TLX impedes the progression of TNBC. Several ligands have been shown to regulate the transcriptional activity of TLX, providing a framework for the future development of this receptor for therapeutic intervention.

Snake fungal disease (SFD; Ophidiomyces ophiodiicola) is posing a significant threat to several free-ranging populations of pitvipers. Triazole antifungals have been proposed for the treatment of mycoses in reptiles; however, data are lacking about their safety and efficacy in snakes with SFD. Study 1 investigated in vitro susceptibility, and identified that plasma concentrations250 ng/ml (voriconazole) and1,000 ng/ml (itraconazole) may be effective in vivo for SFD. In Study 2, the pharmacokinetics after a single subcutaneous voriconazole injection were assessed in apparently healthy free-ranging cottonmouths (Agkistrodon piscivorus). Based on pilot-study results, four snakes were administered a single injection of voriconazole (5 mg/kg). One pilot snake and three full-study snakes died within 12 hr of voriconazole administration. All surviving snakes maintained plasma concentrations250 ng/ml for 12-24 hr. In Study 3, two Eastern massasaugas (Sistrurus catenatus) and a timber rattlesnake (Crotalus horridus horridus) diagnosed with SFD were treated with voriconazole delivered by subcutaneous osmotic pumps. The timber rattlesnake (12.1-17.5 mg/kg/hr) reached therapeutic concentrations, whereas the massasaugas (1.02-1.6 mg/kg/hr) did not. In Study 4, the pharmacokinetics of a single 10-mg/kg per-cloaca dose of itraconazole (Sporanox®) was evaluated in seven apparently healthy free-ranging cottonmouths. Similarly, the plasma and tissue concentrations did not meet therapeutic concentrations based on in vitro data. The data presented in this report serve as an initial step toward understanding the pharmacokinetics, efficacy, and safety of triazole antifungals in pitviper species with SFD. Further study is needed to determine the appropriate dose and route of administration of triazole antifungals in pitviper species.

Ophidiomycosis, historically referred to as snake fungal disease (SFD), caused by

A 5-yr-old, intact male red ruffed lemur ( Varecia rubra ) presented for evaluation as the result of a 1-wk history of lethargy and hyporexia. Physical examination findings included thin body condition, muffled heart sounds, harsh lung sounds, and liquid brown diarrhea. Complete blood count and serum biochemistry showed an inflammatory leukogram, mild hyponatremia, and mild hypochloremia. Orthogonal trunk radiographs revealed a severe alveolar pattern in the right cranial lung lobes with cardiac silhouette effacement. Thoracic ultrasound confirmed a large, hypoechoic mass in the right lung lobes. Fine-needle aspiration of the lung mass and cytology revealed fungal yeast organisms, consistent with Blastomyces dermatitidis. Blastomyces Quantitative EIA Test on urine was positive. Postmortem examination confirmed systemic blastomycosis involving the lung, tracheobronchial lymph nodes, spleen, kidney, liver, cerebrum, and eye. To the authors' knowledge, this is the first report of blastomycosis in a prosimian species.

Determine the extent to which pre-fitting acceptable noise level (ANL), with or without other predictors such as hearing-aid experience, can predict real-world hearing-aid outcomes at three and 12 months post-fitting.ANLs were measured before hearing-aid fitting. Post-fitting outcome was assessed using the international outcome inventory for hearing aids (IOI-HA) and a hearing-aid use questionnaire. Models that predicted outcomes (successful vs. unsuccessful) were built using logistic regression and several machine learning algorithms, and were evaluated using the cross-validation technique.A total of 132 adults with hearing impairment.The prediction accuracy of the models ranged from 61% to 68% (IOI-HA) and from 55% to 61% (hearing-aid use questionnaire). The models performed more poorly in predicting 12-month than three-month outcomes. The ANL cutoff between successful and unsuccessful users was higher for experienced (∼18 dB) than first-time hearing-aid users (∼10 dB), indicating that most experienced users will be predicted as successful users regardless of their ANLs.Pre-fitting ANL is more useful in predicting short-term (three months) hearing-aid outcomes for first-time users, as measured by the IOI-HA. The prediction accuracy was lower than the accuracy reported by some previous research that used a cross-sectional design.

Background:Voriconazole is a broad-spectrum azole exhibiting strong anti-Aspergillus activity and good long-term tolerance. However, the evidence for voriconazole efficacy against refractory Aspergillus otomycosis is weak.Method:We reviewed the medical records of patients with Aspergillus otomycosis treated with voriconazole from January 2008 to June 2012 in a Taiwanese regional hospital. Demographic data and information regarding underlying diseases, clinical features, treatment and outcome were assessed.Results:In total, 14 cases of Aspergillus otomycosis were treated with voriconazole, including 5 patients with Aspergillus invasive otitis externa. All patients had failed to respond to local treatment, antibiotics or topical agents. One case was lost to follow up. The symptoms of two patients recurred after voriconazole treatment: one patient received a second 12-week course of voriconazole and was cured; and symptoms of the other patient recurred after a second 12-week course of voriconazole, leading to surgical debridement. The remaining 11 patients were cured by voriconazole treatment without extensive surgical debridement.Conclusion:This study demonstrates that voriconazole can be a very effective and convenient therapeutic option for the management of refractory Aspergillus otomycosis.

The objective of the present study was to characterize and compare the differences in gene expression associated with innate immune response of blood monocytes (Mos) between healthy subclinically PCV2-infected and PCV2-free pigs prior to and after lipopolysaccharide (LPS) stimulation in vitro by relative quantitative real-time PCR (q-rt-PCR). Genes coding for 24 innate molecules, including toll-like receptors (TLRs), interferon-regulatory factors (IRFs), nuclear factor kappa-light chain-enhancer of activated B cells (NF-κB) and pro-inflammatory cytokines were evaluated. When compared with PCV2-free pigs, Mos from subclinically PCV2-infected pigs showed significantly lower mRNA expression levels in TLR-9, IRF-3, IRF-6, IRF-7, IL-6, IL-12p35, IL-12p40 and IFN-α under no further stimulation. Following LPS stimulation in vitro, a broad and/or obvious reduction in TLRs, IRFs, IL-12 and IFN-α along with increase in IL-1α, IL-6, IL-8, IL-10 and/or TNF-α were seen in both PCV2-free and subclinically PCV2-infected pigs; when compared with PCV2-free pigs, the subclinically PCV2-infected pigs had significantly higher expression levels in TLR-10 and IRF-1 but lower expression levels in IRF-6, IL-1α and IL-12p40. On the contrary, the expression level of NF-κB was consistently higher in subclinically PCV2-infected pigs than in PCV2-free pigs with or without LPS stimulation. The changes seen in the present study suggest that the subclinically PCV2-infected pigs may look healthy clinically, but their innate immunity has become dis-regulated or is in an improper status. The adverse condition may become even worse when exposed to certain bacterial products such as endotoxin. Such alterations in the innate immune system may make the subclinically PCV2-infected pigs more vulnerable to the secondary infection and subsequent PCV2-associated disease development.

Ophidiomycosis, historically referred to as snake fungal disease (SFD), caused by Ophidiomyces ophiodiicola, is a significant disease of snakes characterized by crusty scales, pustules, subcutaneous nodules, and death. Ophidiomycosis is a proposed threat to sustainability of free-ranging snake populations throughout the United States and Europe, but the clinical progression during periods of reproductive activity (gravid females, neonates) is unknown. In spring 2012, five apparently healthy gravid eastern massasauga (Sistrurus catenatus) rattlesnakes from Clinton County, Illinois, were brought into captivity to give birth and be returned into the population. While in captivity, one adult female and 21 neonates died. Five individuals were subsequently confirmed positive for O. ophiodiicola by using quantitative polymerase chain reaction (qPCR). In 2016, a gravid timber rattlesnake (Crotalus horridus) with ophidiomycosis from Jackson County, Illinois, gave birth in captivity to 13 neonates. Skin swabs were taken from all neonates immediately after birth and confirmed negative for O. ophiodiicola by using qPCR. The neonates remained housed with the positive female for 10 days before all animals were reswabbed and released back into the wild. One neonate was O. ophiodiicola positive at time of release. The initial negative result followed by a positive result several days postpartum suggests that the neonate was infected by the female after direct contact. Both case series represent natural infection of neonates after parturition and highlight the importance of this disease in a demographically important age class.

The objective of this prospective study was to investigate the relationship between acceptable noise level (ANL), which was measured using Taiwanese and the international speech test signal (ISTS), and real-world hearing-aid success for listeners who were representative of the population commonly seen in clinics.Unaided ANLs were measured pre-hearing-aid fitting. Hearing-aid success was assessed three months post-fitting using the international outcome inventory for hearing aids (IOI-HA) and a hearing-aid use questionnaire.Eighty adults with hearing impairment completed the study.Both Taiwanese and ISTS ANLs were significantly associated with hearing-aid success, with higher ANLs suggesting poorer outcomes. However, the ANL's prediction accuracy for the probability of hearing-aid success was either much lower than that suggested by some literature, or was not much different from that of simply predicting all listeners as successful users.The current study suggested the possibility of using ANL to predict hearing-aid success. However, the usefulness of ANL as a clinical tool is unlikely to be as great as indicated by the literature.

Acceptable noise level (ANL) determines the maximum noise level that a listener is willing to accept while listening to speech. The objective of this study was to determine the equivalence of ANL measured using different speech stimuli for native speakers who lived in the U.S. and Taiwan.ANLs were measured using English, Mandarin, and the international speech test signal (ISTS) at each site. The same babble noise was used across speech stimuli. The ANLs were considered equivalent if the difference was unlikely to be greater than 3 dB.Thirty adults with normal hearing were recruited at each site.For each site, the equivalence test suggested that the native-language and foreign-language ANLs were equivalent. Between the two sites, ANLs measured using the listener's native language were also equivalent. Although the ISTS ANL obtained within each site was equivalent to, and highly correlated to, the native-language ANL, the data were unable to confirm the equivalence of the ISTS ANLs obtained from the two sites.The results suggested the possibility of directly comparing ANL measures carried out in different countries using different languages. However, it remains unclear if the ISTS can serve as an international ANL stimulus.

Background: The aim of this study was to determine the influence of inflammation on acute phase protein and epithelial-mesenchymal transition (EMT) in buccal cancer. Methods: Western blotting was carried out to investigate the expression of haptoglobin and epithelial-mesenchymal transition in oral cancer cell lines with or without IL-6 stimulation. We studied patients with buccal cancer patients without distant metastasis at diagnosis. Correlation between cellular haptoglobin, EMT, and clinical characteristics of buccal cancer was analyzed to assess the prognostic value of cellular haptoglobin level and EMT. The relationship of haptoglobin, and EMT expression with survival was assessed using Cox proportional hazard models. Results: Western blotting analysis showed that increased haptoglobin protein was associated with overexpression of vimentin. Under IL-6 stimulation, overexpression of haptoglobin, EMT-associated motile phenotype was noted in OC2 cell lines. Overexpression of haptoglobin was also associated with an increased risk for locoregional recurrence [hazard ratio (HR) 1.04; p=0.011] after adjusting for age, gender, disease site, stage, and treatment modality. Conclusions: Increased cellular expression of haptoglobin is associated with EMT in oral cancer cell lines and this phenomenon could be exaggerated with IL-6. Cellular expression of haptoglobin is related to locoregional recurrence rate in buccal cancer patients.

Dendritic cells (DCs) are professional antigen presenting cells cooperating with other immune cells for the activation of innate and adaptive immune responses. The objective of the present study was to investigate the replication activity of porcine circovirus type 2 (PCV2) in DCs and/or lymphocytes during their cross talk and its possible mechanism. Two models were set, herein. Swine blood monocyte (Mo)-derived DCs (MoDCs) or peripheral blood lymphocytes (PBLs) were inoculated with PCV2 prior to their co-cultivation. Bacterial lipopolysaccharide (LPS) and concanavalin A (Con A) were used to stimulate MoDCs and PBLs, respectively. During 6 days of cultivation, a high PCV2 antigen-containing rate without detectable intranuclear signals and a slight but significant increase in the copy number of PCV2 genome were detected in PCV2-inoculated MoDCs. The presence of LPS alone or PCV2-free PBLs, however, had no effect on the location of PCV2 antigens or copy number of PCV2 genome in PCV2-inoculated MoDCs. On the contrary, active PCV2 replication occurred in Con A-stimulated PCV2-inoculated PBLs. When compared with blood Mos, MoDCs induced significantly higher cell proliferation and intensified PCV2 replication in Con A-stimulated PCV2-inoculated PBLs, for which direct contact between MoDCs and lymphocytes was required. Among the cytokines secreted by Con A-activated PBLs, interleukin (IL)-2, but not IL-4 or interferon-γ, could induce cell proliferation and PCV2 replication in PCV2-inoculated PBLs. The findings suggest that although MoDCs support only limited PCV2 replication in themselves, their accessory cell function is required for cell proliferation and PCV2 replication in PCV2-infected lymphocytes.

Olfactory neuroblastoma, also known as esthesioneuroblastoma, is a rare malignant head and neck tumor. Olfactory neuroblastoma accounts for approximately 3–5% of all malignant nasal tumors. We present a case of olfactory neuroblastoma in a 61-year-old man who underwent endoscopic surgery followed by postoperative irradiation. Right neck metastasis was diagnosed about 6 months after endoscopic surgery. Supraomohyoid dissection of the right side of the neck was performed, but right parotid metastasis was identified about 3 months later. Boost radiotherapy was applied to the right metastatic parotid. There was no evidence of recurrence on head and neck magnetic resonance imaging and endoscopic examination during the 2-year follow-up.

A loop-mediated isothermal amplification (LAMP) method with a real-time monitoring system was developed for the detection of porcine circovirus type 1 (PCV1) in commercial swine vaccines. This method was highly specific for PCV1. No cross-reaction to porcine circovirus type 2, porcine parvovirus, pseudorabies virus, classical swine fever virus, and porcine reproductive and respiratory syndrome virus was observed. The analytical sensitivity of the LAMP for PCV1 DNA was 10 copies/μl in the case of positive recombinant plasmid comparable to that obtained from the nested polymerase chain reaction (nested PCR). Furthermore, 25 commercial swine vaccines were tested by both the LAMP and the nested PCR, and three of them were tested positive for PCV1 DNA. These results indicate that PCV1 DNA can be real-time detected by the LAMP; the method was highly specific, sensitive, and rapid for the detection of PCV1 DNA, particularly in commercial swine vaccines.

The immunopathogenesis of porcine circovirus type 2 (PCV2) infection in conventional pigs is complicated by various environmental factors and individual variation and is difficult to be completely reproduced experimentally. In the present field-based study, a tissue microarray (TMA) consisting of a series of lymphoid follicles having different PCV2-loads was constructed using formalin-fixed and paraffin-embedded superficial inguinal lymph nodes (LNs) from 102 pigs. Using the TMA, a wide range of parameters, including co-infected viral pathogens, immune cell subsets, and cell apoptosis/proliferation activity by immunohistochemical (IHC) staining or in situ hybridization (ISH) were measured, characterized, and compared. The signal location and area extent of each parameter were interpreted by pathologists, semi-quantified by automated image analysis software, and analyzed statistically. The results herein demonstrated a significant negative correlation between PCV2 and CD79a (p < 0.001) and a significant positive correlation between PCV2 and lysozyme (p < 0.001) or TUNEL (p < 0.001) using Pearson correlation analysis. The amount of porcine respiratory and reproductive syndrome virus (PRRSV) and porcine parvovirus antigens did not correlate with the tissue loads of PCV2 nucleic acid. Multiple regression analysis further predicted that PCV2 contributed major effects on CD79a, lysozyme, and TUNEL but PRRSV showed relatively less effects on these parameters. In addition, the total signal intensity of Ki67 (index of cell proliferation activity) did not change significantly among cases with different PCV2 loads; however, as the loading of PCV2 nucleic acid increased, the main contribution of Ki67 signal gradually shifted from B cells in the germinal center to T cells and macrophages in the interfollicular regions. In the present study, the use of TMA to establish a mathematical model with a wider range of statistical analysis can bring us a step forward to understand the immunopathogenesis of PCV2 infection-associated follicular changes in LNs.

The aim of the study was to evaluate the immunogenicity of the ORF5-encoded major envelop glycoprotein 5 (GP5) of porcine reproductive and respiratory syndrome virus (PRRSV) expressed in tobacco plant as a potential pig oral vaccine in protection against PRRSV infection. Six-week-old PRRSV-free pigs were fed four times orally with 50g of chopped fresh GP5 transgenic tobacco leaves (GP5-T) (GP5 reaching 0.011% of total soluble protein) or wild-type tobacco leaves (W-T) each on days 0, 14, 28, and 42. Samples of serum, saliva, and peripheral blood mononuclear cells (PBMCs) were collected on days -1, 6, 13, 20, 27, 34, 41, and 48 after the initial oral vaccination. A similar vaccination-dependent gradual increase in the responses of serum and saliva anti-PRRSV total IgG and IgA, respectively, and in the levels of PRRSV-specific blastogenic response of PBMCs was seen in GP5-T-treated pigs; all statistically significant elevations occurred after the 2nd vaccination and were revealed after 20 days post-initial oral vaccination (DPIOV). Pigs fed on GP5-T also developed serum neutralizing antibodies to PRRSV at a titer of 1:4-1:8 after the 4th vaccination by 48 DPIOV. No detectable anti-PRRSV antibody responses and PRRSV-specific blastogenic response were seen in W-T-treated pigs. The present study has demonstrated that pigs fed on GP5-T could develop specific mucosal as well as systemic humoral and cellular immune responses against PRRSV. The results also support that transgenic plant as GP5-T can be an effective system for oral delivery of recombinant subunit vaccines in pigs.

Taking advantage of the high sensitivity of polymerase chain reaction (PCR) and the cell-localizing ability of in situ hybridization (ISH), an indirect in situ PCR (ISPCR) method was developed for detecting the distribution of porcine circovirus type 2 (PCV2) in formalin-fixed and paraffin-embedded inguinal lymph nodes obtained from clinically healthy PCV2-carrier pigs and postweaning multisystemic wasting syndrome (PMWS)-affected pigs. Comparisons of the relative sensitivity of indirect ISPCR with other routinely used diagnostic methods for PCV2 indicated that nested PCR was the most sensitive method followed by indirect ISPCR, conventional PCR, ISH, and immunohistochemical (IHC) staining. Although indirect ISPCR, ISH, and IHC staining all revealed a similar signal distribution pattern of PCV2, using indirect ISPCR allowed specific amplification and detection of previously uneasily detected PCV2 signal than by routine ISH or IHC staining, particularly in those cells within the germinal center in clinically healthy PCV2-carrier pigs. Furthermore, six different PCV2 signal expression patterns in conjunction with the correlated lymphoid lesion stages were classified to describe the tissue morphological changes and viral infection. The result indicates that indirect ISPCR is a more effective, cell-based diagnostic tool with good specificity to detect limited PCV2 infection in formalin-fixed and paraffin-embedded tissue specimens and it would be a useful tool for further exploring the pathogenesis of PCV2 infection.

Changes in tumor necrosis factor-alpha (TNF-alpha) and soluble interleukin-2 (sIL-2R) levels appear to be closely related to tumor progression and prognosis in nasopharyngeal carcinoma (NPC). Further investigation is suggested.The study examined whether changes in TNF-alpha and sIL-2R in NPC can be used to predict tumor progression and prognosis.The study was carried out in 58 patients with NPC newly diagnosed from December 2003 to December 2006 at a single institution and 60 control subjects of comparable age. Blood levels of TNF-alpha and sIL-2R were monitored before, during, and 3 months and 1 year after treatment.Differences in TNF-alpha level between patients with NPC in all four stages and healthy controls and in sIL-2R level between patients with advanced stage NPC and healthy controls were significant (p0.05). Furthermore, 1 year after completing radiotherapy, levels of TNF-alpha and sIL-2R in patients with recurrent tumors were significantly different from those in patients without recurrence and healthy control subjects.

Objectives To elucidate the effect of primary tumor volume (PTV) on treatment outcomes in patients with nasopharyngeal carcinoma (NPC) who were treated with radiotherapy or concurrent chemoradiotherapy. The TNM staging system developed by American Joint Committee of Cancer (AJCC) is universally used and accepted but its prediction of prognosis in NPC receives a lot of challenge. Primary tumor volume had been reported to have close relationship with prognosis of head and neck cancer. We may predict prognosis of NPC with PTV. Methods From 1999 to 2006, 91 patients with newly diagnosed NPC who were treated with radiotherapy or CCRT were enrolled in the study. Computed tomography-derived or magnetic resonance-derived primary tumor volume was calculated. The correlation between AJCC disease stage, primary tumor volume and disease-specific survival were analyzed. Multivariate analyses using the Cox proportional hazard model was performed. Results The median primary tumor volume for the whole series was 11.39 ml (range 1.25–166.58 ml). The median primary tumor volume was 2.69 ml in T1 disease, 10.14 ml in T2 disease, 15.41 ml in T3 disease, and 26.69 ml in T4 disease. Hazard ratio increased with tumor volume, ranging from 5.91 (95% confidence interval (95% CI) 1.13–26.43) for tumor volumes between 20 ml and 40 ml, and 15.72 (95% CI, 3.82–61.05) for tumor volumes >40 ml. With both tumor volume and T classification in the same Cox regression model, only tumor volume remained statistically significant in the prognosis of NPC. Conclusion Primary tumor volume has closer relationship with survival rates of patients with NPC. Calculation of primary tumor volume may be further considered to improve the current staging system.

A weekly regimen of cisplatin 30 mg/m2 as concurrent chemoradiation (CCRT) was effective in elderly nasopharyngeal carcinoma (NPC) patients, with acceptable and reversible acute toxicity following CCRT therapy.The purpose of this retrospective study was to evaluate the efficacy, toxicity, and tolerability of a multi-modal treatment strategy in elderly NPC patients. Subsequent systemic adjuvant chemotherapy was administered to achieve systemic control.From December 2002 to December 2006, 26 NPC patients over 60 years of age who had stage IIB to IV NPC were evaluated in this retrospective analysis. The CCRT chemotherapy protocol consisted of eight weekly doses of cisplatin 30 mg/m2 administered in an outpatient setting. Adjuvant systemic chemotherapy consisted of four monthly cycles of cisplatin (20 mg/m2/day) plus 5-fluorouracil (1000 mg/m2/day) for 5 consecutive days. Study end points included treatment outcome, compliance, and toxicity.The 2-year overall survival, disease-free survival, local control, and distant metastasis-free rate were 87%, 73%, 92%, and 76%, respectively. Over 80% of patients were able to take more than six doses of weekly cisplatin during CCRT; however, nearly half of the patients had grade 3 hematological toxicity during adjuvant therapy requiring treatment modification or cessation of further adjuvant therapy.

Fumonisin B1is a mycotoxin that causes lethal pulmonary edema in swine. Sphinganine, sphingosine, and the sphinganine to sphingosine ratio are important biomarkers for fumonisin B1exposure. Currently, tissues selected for sphinganine and sphingosine analyses are frozen at −80°C until analyses take place. However, for diagnostics and some research projects, formalin is used more routinely as a preservative for long-term storage of tissues. To determine whether formalin-fixed tissues could be used for sphinganine and sphingosine analyses, sphinganine and sphingosine concentrations were quantified in both frozen and formalin-fixed lung, liver, kidney, and heart from fumonisin B1-treated and control pigs. Tissues were evaluated 3 months after freezing and 3, 6, and 12 months after formalin fixation. Sphinganine, sphingosine, and the sphinganine to sphingosine ratio of both frozen and formalin-fixed lung and liver from fumonisin B1-treated pigs were elevated. Formalin-fixed tissues had lower sphinganine and sphingosine concentrations but higher sphinganine to sphingosine ratios than the corresponding frozen tissues. Storage in formalin for up to 12 months did not affect the results. Sphingosine analysis could not be performed in formalin-fixed heart and kidney because of noninterpretable chromatograms. Therefore, formalin-fixed lung and liver can be used to determine fumonisin B1-induced sphinganine and sphingosine alterations in swine, with the sphinganine to sphingosine ratio being the most useful.

Fumonisin mycotoxicosis in pigs causes a decrease in mean aortic pressure, an increase in mean pulmonary arterial pressure, and increases in serum concentrations of sphinganine (3.2 microM) and sphingosine (1.4 microM). To determine a causal relationship between the hemodynamic changes and sphingolipid alterations, we examined the in vitro effects of sphinganine, sphingosine, and sphingosine-1-phosphate on porcine aortic and pulmonary arterial rings. Both sphinganine and sphingosine relaxed un-contracted and phenylephrine-contracted aortic rings at > or = 10 microM and > or = 1 microM, respectively. Sphingosine (> or = 10 microM) relaxed un-contracted and phenylephrine-contracted pulmonary arterial rings, whereas sphingosine-1-phosphate (10 microM) contracted pulmonary arterial rings. Sphingosine (3 microM) also impaired the contractile response of pulmonary artery rings to 60 mM KCl. The results suggested that the systemic hypotension caused by fumonisin is mediated, in part, by increases in serum sphinganine and sphingosine concentrations, and the pulmonary hypertension is mediated, in part, by increased sphingosine-1-phosphate concentrations.

Escherichia coli heat-labile enterotoxin B subunit (LTB) can be used as an adjuvant for co-administered antigens. Our previous study showed that the expression of neutralizing epitope GP5 of porcine reproductive and respiratory syndrome virus (PRRSV) in transgenic tobacco plant (GP5-T) could induce PRRSV-specific immune responses in pigs. A transgenic tobacco plant co-expressing LTB and PRRSV GP5 as a fusion protein (LTB-GP5-T) was further constructed and its immunogenicity was evaluated. Pigs were given orally three consecutive doses of equal concentration of recombinant GP5 protein expressed in leaves of LTB-GP5-T or GP5-T at a 2-week interval and challenged with PRRSV at 7 weeks post-initial immunization. Pigs receiving LTB-GP5-T or GP5-T developed PRRSV-specific antibody- and cell-mediated immunity and showed significantly lower viremia and tissue viral load and milder lung lesions than wild type tobacco plant (W-T). The LTB-GP5-T-treated group had relatively higher immune responses than the GP5-T-treated group, although the differences were not statistically significant.

Background Current staging systems have limited ability to adjust optimal therapy in advanced nasopharyngeal carcinoma (NPC). This study aimed to delineate the correlation between tumor volume, treatment outcome and chemotherapy cycles in advanced NPC. Methods A retrospective review of 110 patients with stage III-IV NPC was performed. All patients were treated first with neoadjuvant chemotherapy, then concurrent chemoradiation, and followed by adjuvant chemotherapy as being the definitive therapy. Gross tumor volume of primary tumor plus retropharyngeal nodes (GTVprn) was calculated to be an index of treatment outcome. Results GTVprn had a close relationship with survival and recurrence in advanced NPC. Large GTVprn (≧13 ml) was associated with a significantly poorer local control, lower distant metastasis-free rate, and poorer survival. In patients with GTVprn ≧ 13 ml, overall survival was better after ≧4 cycles of chemotherapy than after less than 4 cycles. Conclusions The incorporation of GTVprn can provide more information to adjust treatment strategy.

The heterodimer of glycoprotein 5 (GP5) and non-glycosylated matrix protein (M) is the leading target for the development of new generation of vaccines against porcine reproductive and respiratory syndrome virus (PRRSV) infection. It has been demonstrated that DNA vaccine co-expressing GP5 and M proteins as a fusion protein aroused better immunogenicity than that expressing GP5 or M alone, but it was no better than the DNA vaccine co-expressing GP5 and M proteins with two different promoters. Altered natural conformation of the co-expressed GP5 and M fusion protein was considered as the major cause. Glycine-proline-glycine-proline (GPGP) linker can minimize the conformational changes in tertiary structure and provide flexibility of the peptide chain. The objective of this study was to evaluate whether the immunogenicity of DNA constructs co-expressing GP5 and M proteins linked by GPGP could be enhanced in pigs. Three recombinant DNA constructs expressing GP5/M fusion protein without GPGP linker (pcDNA-56), GP5/M fusion protein conjugated by GPGP linker (pcDNA-5L6), and M/GP5 fusion protein conjugated by GPGP linker (pcDNA-6L5) were established. Sixteen PRRSV-free pigs were randomly assigned to four groups and inoculated intramuscularly with 3 consecutive doses of 500 μg of empty vector pcDNA3.1, pcDNA-56, pcDNA-5L6 or pcDNA-6L5 each at a 2-week interval followed by challenge with 5 × 10(5) TCID(50) PRRSV at 3 weeks after the final inoculation. All pcDNA-56-, pcDNA-5L6-, and pcDNA-6L5- but not pcDNA-3.1-inoculated pigs developed neutralizing antibodies (NAs) 3 weeks after the final inoculation and a gradual increase in NA titers after PRRSV challenge, indicating that pigs inoculated with these DNA constructs could establish a sufficient immune memory. The pcDNA-5L6- and pcDNA-6L5-inoculated pigs displayed lower level and shorter period of viremia and lower tissue viral load following PRRSV challenge than did the pcDNA-56-inoculated pigs. The strategy of co-expressing GPGP-linked GP5 and M fusion protein may be a promising approach for future PRRSV vaccine development, possibly via the improvement of natural conformation of the target fusion protein.

International audience; Granulomatous lymphadenitis is one of the pathognomonic lesions in post-weaning multisystemic wasting syndrome (PMWS)-affected pigs. This unique lesion has not been reported in direct association with viral infection in pigs. The objective of the present study was to evaluate whether porcine circovirus type 2 (PCV2) alone is able to induce functional modulation in porcine monocytic cells in vitro to elucidate its possible role in the development of granulomatous inflammation. It was found that the proliferation activity of blood monocytes (Mo) and monocyte-derived macrophages (MDM) was significantly enhanced by PCV2. During monocyte-macrophage differentiation, the PCV2 antigen-containing rate and formation of multinucleated giant cells (MGC) were significantly increased in MDM when compared to those in Mo. The MDM-derived MGC displayed a significantly higher PCV2 antigen-containing rate than did the mono-nucleated MDM. Supernatants from PCV2-inoculated MDM at 24 h post-inoculation induced an increased tendency of chemotactic activity for blood Mo. At the same inoculation time period, levels of mRNA expression of the monocytic chemokines, monocyte chemoattractant protein-1 and macrophage inflammatory protein-1, also significantly increased in PCV2-inoculated MDM. The results suggest that PCV2 alone may induce cell proliferation, fusion, and chemokine expression in swine monocytic cells. Thus, PCV2 itself may play a significant role in the induction of granulomatous inflammation in PMWS-affected pigs.

Amoebiasis is one of the most common protozoal diseases of reptiles, but amoebic myositis has not been reported in any animal species. An 11-year-old, male common water monitor lizard (Varanus salvator) was found dead with several subacute ulcerated skin wounds. Gross examination revealed multiple discrete to coalescing, white-yellow to gray, caseous foci scattered in the skeletal muscles and liver. The mucosa of small intestine was thickened, red, and contained many variably sized, dark red ulcers, with depressed and hemorrhagic centers. Histopathologic examination revealed severe necrotizing and granulomatous myositis, hepatitis, and enteritis accompanied by large numbers of intralesional, 10-20-μm diameter, periodic acid-Schiff-positive, amoeboid protozoa. Gene sequence analysis of a 136-bp region of the 18S ribosomal RNA amplified by polymerase chain reaction revealed 98–100% similarity with Entamoeba invadens. Aside from intestinal and hepatic involvement, no other internal organs were affected. The muscular infection by E. invadens likely resulted from a combination of direct invasion of trophozoites via skin wounds and hematogenous spread.

We suggest that concurrent chemoradiation (CCRT) is an effective definitive treatment for patients with advanced hypopharyngeal carcinoma who are unfit for or refuse surgery. A high dose of radiation (70 Gy) should be given to achieve acceptable local control rates and survival.The purpose of this retrospective study was to compare the treatment results of locally advanced hypopharyngeal carcinoma with two different protocols.From December 1995 to December 2004, 74 patients with locally advanced hypopharyngeal cancer were treated with CCRT or surgery plus postoperative radiotherapy (SRT). Their treatment results were reviewed by retrospective analysis. The study points included outcome, toxicity, and prognostic factors.There was no significant difference in T and N status between the two treatment groups, nor were there significant differences in overall or disease-free survival or the incidence of distant metastasis (p0.05). In the CCRT group and SRT group, the estimated 3-year overall survival was 39% and 44%, respectively. The SRT group had better local control than the CCRT group (p0.05). Relatively, 27% patients retained their larynx function for more than 2 years in the CCRT group. Radiation doses70 Gy yielded significantly better survival and local control than doses70 Gy (p0.05).

Conclusions. Young patients with squamous cell carcinoma (SCC) of the oral tongue developed fewer locoregional recurrences. The overall survival and disease-specific survival rates were better in the young patient population. Objectives. To compare the survival rates of patients under 45 years of age and diagnosed with SCC of the oral tongue with those of patients older than 45 years. Patients and methods. A retrospective review of 20 patients under 45 years of age with SCC of the oral tongue was performed. These patients were matched to an older population by sex and clinical stage. Overall survival, disease-free survival, disease-specific survival, and rates of local, regional and distant metastases were determined for both populations. Results. Stage and treatment modality were similar in the two age groups. There were significant differences in overall survival (p=0.013) and disease-specific survival (p=0.046) favoring young patients. Rates of locoregional recurrence and distant metastasis were higher...

There is an increased incidence of oral squamous cell carcinoma (SCC) in young patients. However, the literature concerning prognosis for young patients with oral SCC is conflicting. The aim of this study was to compare the survival rates of patients under 45 years of age and diagnosed with SCC of the oral cavity with those of patients older than 45 years. A retrospective review of 28 patients under 45 years of age with SCC of the oral cavity was performed. We matched each of these patients by sex, tumor site, and overall stage with two patients older than 45 years. Overall survival, disease-free survival, disease-specific survival, rates of local, regional and distant metastasis were determined for both populations. Survival analysis was performed using Cox proportional hazard models. Stage and treatment modality were similar in the two groups. There were significant differences in overall survival (P=0.024) and disease-specific survival (P=0.03) favoring young patients. Locoregional recurrence and distant metastasis were higher in the older patients. Furthermore, matched survival analysis demonstrated a difference in overall survival rate [risk ratio (RR) 2.457; P=0.033] and disease-specific survival [RR 2.475; P=0.048]. Young patients with SCC of the oral cavity developed fewer locoregional recurrences and distant metastasis. The overall survival and disease-specific survival rates were better in the young patient population.

BACKGROUND: To compare the infection rates between cetuximab-treated patients with head and neck cancers (HNC) and untreated patients. METHODOLOGY: A national cohort of 1083 HNC patients identified in 2010 from the Taiwan National Health Insurance Research Database was established. After patients were followed for one year, propensity score analysis and instrumental variable analysis were performed to assess the association between cetuximab therapy and the infection rates. RESULTS: HNC patients receiving cetuximab (n = 158) were older, had lower SES, and resided more frequently in rural areas as compared to those without cetuximab therapy. 125 patients, 32 (20.3%) in the group using cetuximab and 93 (10.1%) in the group not using it presented infections. The propensity score analysis revealed a 2.3-fold (adjusted odds ratio [OR] = 2.27; 95% CI, 1.46-3.54; P = 0.001) increased risk for infection in HNC patients treated with cetuximab. However, using IVA, the average treatment effect of cetuximab was not statistically associated with increased risk of infection (OR, 0.87; 95% CI, 0.61-1.14). CONCLUSIONS: Cetuximab therapy was not statistically associated with infection rate in HNC patients. However, older HNC patients using cetuximab may incur up to 33% infection rate during one year. Particular attention should be given to older HNC patients treated with cetuximab.

Sonographic texture analysis can reflect histopathological components and their arrangement in metastatic and common benign lymph nodes. It is helpful in differentiation between metastatic and benign lymph node lesions for target selection during biopsy of multiple lymph nodes and the strategy of the management. Two ultrasound systems, 107 sonographic regions of interest (ROIs) of metastases and 174 sonographic ROIs of common benign lymph nodes, were recruited in the study. Thirteen texture features derived from co-occurrence matrix were used in characterization of above ROI ultrasound images. Support vector machine (SVM) was used as a classifier and a feature selector. The experimental results show that the entropy gains the best cross-validation accuracy of 94.66% and 87.73% in both ultrasound systems 1 and 2 for the classification of metastatic and benign lymph nodes disease. The accuracy can be further increased to 97.86% and 100% by the combination of the sum average in the study. There are significantly higher entropy and sum average values of the metastatic lymph nodes than of the benign lymph nodes, which are due to the heterogeneous compositions and arrangement of larger cancer cells, lymphocytes, and stroma in metastatic lymph nodes that contrast with simple inflammatory cells infiltration in common benign lymph nodes.

ObjectiveThe aim of this study was to retrospectively analyze locally advanced buccal cancer in order to determine tumor characteristics and any other prognostic factors that may influence the survival of patients with the disease.Patients and MethodsThe records of 54 patients with stage III to IV locally advanced buccal cancer (TNM system) who were seen from August 2000 to June 2008 at one institution were reviewed. The patients received radical surgery and then adjuvant treatment. The adjuvant treatment in cluded concurrent chemoradiotherapy or radiotherapy alone. The chemotherapy protocol consisted of two 4-weekly courses of concurrent cisplatin and 5-fluorouracil followed by another two 4-weekly courses after radiotherapy, with regimens of cisplatin (60–100 mg/m2/day) on day 1 and 5-fluorouracil (1000 mg/m2/day) on days 1–5.ResultsThe 3-year cumulative overall survival, disease-specific survival, disease-free survival, locoregional control, and distant metastasis-free survival rates were 66%, 69%, 58%, 65%, and 92%, respectively. Univariate analysis indicated that the pN classification, the surgical margin, and the presence of extracapsular spread were significantly associated with overall survival. On multivariate analysis, pN classification and surgical margin significantly affected disease-free survival. The pN classification (pN0 vs. pN1–3) and the surgical margin (< 1 vs.≥ 1 mm) were the two most significant factors affecting clinical outcome.ConclusionThe presence of lymph node involvement, the presence of extracapsular spread and a surgical margin < 1 mm were strong prognostic factors that were associated with tumor control. More aggressive postoperative therapy is suggested for patients with buccal mucosa carcinoma if these factors exist.

The clinical features of tracheal injury include hoarseness, subcutaneous emphysema and edema, and bruising of the neck. Patients with tracheal injury may present with minimal signs that are easily overlooked. The priority in treating such patients is to appropriately identify the tracheal anatomy before further treatment is administered. In this case, a male patient aged 17 years was injured over the first tracheal ring while riding a motorcycle. His stable condition obscured pathognomonic signs of complete cervical tracheal transection, which caused an unnecessary delay in treatment. In a case such as ours, the application of high resolution computed tomography (CT), as an adjuvant to bronchoscopy is important. If tracheal transection is suspected, the proximal and distal ends of the trachea might be not on the same axis and traditional laryngoscopy should not be performed in haste. A CT virtual bronchoscopic evaluation can provide great accuracy when analyzing the central and segmental bronchi, and can prevent a delayed diagnosis of complete tracheal transection. If the diagnosis is still not definite, then diagnostic bronchoscopy and intubation at the operating room is recommended.