Chimaeric plant-produced bluetongue virus particles as potential vaccine candidates.

Bluetongue virus (BTV) causes bluetongue disease in ruminants and sheep. The current live attenuated and inactivated vaccines available for prevention pose several risks, and there is thus a need for vaccines that are safer, economically viable, and effective against multiple circulating serotypes. This work describes the development of recombinant virus-like particle (VLP) vaccine candidates in plants, which are assembled by co-expression of the four BTV serotype 8 major structural proteins. We show that substitution of a neutralising tip domain of BTV8 VP2 with that of BTV1 VP2 resulted in the assembly of VLPs that stimulated serotype-specific antibodies as well as virus-specific neutralising antibodies.

Role of Intrinsic (Graft) Versus Extrinsic (Host) Factors in the Growth of Transplanted Organs Following Allogeneic and Xenogeneic Transplantation.

In our studies of life-supporting α-1,3-galactocyltransferase knockout (GalT-KO) pig-to-baboon kidneys, we found that some recipients developed increased serum creatinine with growth of the grafts, without histological or immunological evidence of rejection. We hypothesized that the rapid growth of orthotopic pig grafts in smaller baboon recipients may have led to deterioration of organ function. To test this hypothesis for both kidneys and lungs, we assessed whether the growth of outbred (Yorkshire) organ transplants in miniature swine was regulated by intrinsic (graft) or extrinsic (host environment) factors. Yorkshire kidneys exhibited persistent growth in miniature swine, reaching 3.7 times their initial volume over 3 mo versus 1.2 times for miniature swine kidneys over the same time period. Similar rapid early growth of lung allografts was observed and, in this case, led to organ dysfunction. For xenograft kidneys, a review of our results suggests that there is a threshold for kidney graft volume of 25 cm3 /kg of recipient body weight at which cortical ischemia is induced in transplanted GalT-KO kidneys in baboons. These results suggest that intrinsic factors are responsible, at least in part, for growth of donor organs and that this property should be taken into consideration for growth-curve-mismatched transplants, especially for life-supporting organs transplanted into a limited recipient space.

Outcomes with multimodal therapy for elderly patients with rectal cancer.

BACKGROUND: Treatment guidelines for stage II and III rectal cancer include neoadjuvant chemoradiotherapy, surgery and postoperative adjuvant chemotherapy. Although data support this recommendation in younger patients, it is unclear whether this benefit can be extrapolated to elderly patients (aged 75 years or older). METHODS: This was a retrospective review of patients aged at least 75 years with stage II or III rectal cancer who underwent surgery with curative intent from 1996 to 2013 at the Mayo Clinic. Kaplan-Meier analysis and log rank test were used to compare overall survival between therapy groups. Cox proportional hazards model was used to estimate the independent effect of treatment group on survival. RESULTS: A total of 160 elderly patients (median age 80 years) with stage II (66) and stage III (94) rectal cancer underwent surgical resection. Only 30·0 and 33·8 per cent received neoadjuvant or adjuvant therapy respectively. Among patients with stage II disease, there was no significant difference in 60-month survival between patients who received any additional therapy and those who had surgery alone (55 versus 38 per cent respectively; P = 0·184), whereas additional therapy improved survival in patients with stage III tumours (58 versus 30 per cent respectively; P = 0·007). Multivariable analysis found a survival benefit for additional therapy in elderly patients with stage III disease (hazard ratio 0·58, 95 per cent c.i. 0·34 to 0·98). CONCLUSION: A multimodal approach in elderly patients with stage III rectal cancer improved oncological outcomes.

Synthesis and biological evaluation of 2,6-di(furan-3-yl)anthracene-9, 10-dione as an inhibitor of human protein kinase CK2.

Protein kinase CK2 is an emerging target for the therapeutic intervention in human diseases, in particular in cancer. Inhibitors of this enzyme are at current in clinical trials indicating its drug ability. Here we report on the synthesis of two derivatives of 2,6-diaryl-anthracene-9,10-dione, one of them, 2,6-di(furan-3-yl)anthracene-9,10-dione (3), turned out to be active towards CK2, and ATP competitive with an IC50 value of 2.35 µM and a K(i) value of 1.26 µM. Molecular modeling studies were performed to explain the binding affinity of compound 3 in comparison to emodin. These indicated that unlike emodin, compound 3 was not able to perform a hydrogen bond with Lys68, although the compound fits well in the active site of human CK2α, which explains the difference in the measured affinity between those two compounds.

Abundant triatomines in Texas dog kennel environments: Triatomine collections, infection with Trypanosoma cruzi, and blood feeding hosts.

Triatomine insects are vectors of the protozoan parasite Trypanosoma cruzi- the causative agent of Chagas disease. Chagas disease is endemic to Latin America and the southern United States and can cause severe cardiac damage in infected mammals, ranging from chronic disease to sudden death. Identifying interactions among triatomines, T. cruzi discrete typing units (DTUs), and blood feeding hosts is necessary to understand parasite transmission dynamics and effectively protect animal and human health. Through manual insect trapping efforts, kennel staff collections, and with the help of a trained scent detection dog, we collected triatomines from 10 multi-dog kennels across central and south Texas over a one-year period (2018-2019) and tested a subset to determine their T. cruzi infection status and identify the primary bloodmeal hosts. We collected 550 triatomines, including Triatoma gerstaeckeri (n = 515), Triatoma lecticularia (n = 15), Triatoma sanguisuga (n = 6), and Triatoma indictiva (n = 2), with an additional 10 nymphs and 2 adults unable to be identified to species. The trained dog collected 42 triatomines, including nymphs, from areas not previously considered vector habitat by the kennel owners. Using qPCR, we found a T. cruzi infection prevalence of 47 % (74/157), with T. lecticularia individuals more likely to be infected with T. cruzi than other species. Infected insects harbored two T. cruzi discrete typing units: TcI (64 %), TcIV (23 %), and mixed TcI/TcIV infections (13 %). Bloodmeal host identification was successful in 50/149 triatomines, revealing the majority (74 %) fed on a dog (Canis lupus), with other host species including humans (Homo sapiens), raccoons (Procyon lotor), chickens (Gallus gallus), wild pig (Sus scrofa), black vulture (Coragyps atratus), cat (Felis catus), and curve-billed thrasher (Toxostoma curviostre). Given the frequency of interactions between dogs and infected triatomines in these kennel environments, dogs may be an apt target for future vector control and T. cruzi intervention efforts.

Atlas of MRI findings of sacroiliitis in pediatric sacroiliac joints to accompany the updated preliminary OMERACT pediatric JAMRIS (Juvenile Idiopathic Arthritis MRI Score) scoring system: Part I: Active lesions.

Magnetic resonance imaging (MRI) is an increasingly important tool for identifying involvement of the sacroiliac joints (SIJ) in juvenile idiopathic arthritis (JIA). The key feature for diagnosing active sacroiliitis is bone marrow edema (BME), but other features of active arthritis such as joint space inflammation, inflammation in an erosion cavity, capsulitis and enthesitis can be seen as well. Structural changes may also be seen. Systematic MRI assessment of inflammation and structural damage may aid in monitoring the disease course, choice of therapeutics and evaluating treatment response. In this pictorial essay, we illustrate normal MRI findings and growth-related changes of the SIJ in the pediatric population, as well as the different MRI features of SIJ inflammation. This atlas demonstrates fundamental MRI disease features of active inflammation in a format that can serve as a reference for assessing SIJ arthritis according to the updated preliminary JAMRIS (Juvenile Idiopathic Arthritis MRI Score) scoring system proposed by the MRI in JIA working group of Outcome Measures in Rheumatology and Clinical Trials (OMERACT). The atlas is intended to be read in conjunction with its companion Part 2, Structural Lesions.

Atlas of MRI findings of sacroiliitis in pediatric sacroiliac joints to accompany the updated preliminary OMERACT pediatric JAMRIS (Juvenile Idiopathic Arthritis MRI Score) scoring system: Part II: Structural damage lesions.

Magnetic resonance imaging (MRI) is the imaging modality of choice for identifying sacroiliitis in juvenile idiopathic arthritis (JIA). Besides active lesions of sacroiliitis, of which bone marrow edema (BME) is the key feature, structural damage lesions can also be detected. Structural changes include erosion, sclerosis, fat lesion, backfill and ankylosis, and are more common at later stages. Systematic MRI assessment of inflammation and structural damage may aid in monitoring the course of the disease and evaluating treatment options. In this pictorial essay, we illustrate normal MRI findings and growth-related changes of the SIJ in the pediatric population, as well as the different MRI features of structural damage of sacroiliitis. This atlas can serve as a reference for assessing structural lesions of SIJ arthritis according to the updated preliminary JAMRIS (Juvenile Idiopathic Arthritis MRI Score) scoring system proposed by the MRI in JIA working group of Outcome Measures in Rheumatology and Clinical Trials (OMERACT). The atlas is intended to be read in conjunction with its companion Part 1, Active Lesions.

Is p16(INK4a) protein expression in oral ST lesions a reliable precancerous marker?

This study correlates the expression of p16(INK4a) and p53 with the detection of high risk human papillomavirus (HPV) in three clinical grades of smokeless tobacco keratosis (STK) as compared with patients without a history of smokeless tobacco use. Tissue samples, including squamous cell carcinoma (SCC) were evaluated for the expression of p16(INK4a) and p53 by indirect immunohistochemical methods using commercially obtained antibodies. HPV DNA analysis was performed using consensus sequence polymerase chain reaction (PCR). At least focal p16(INK4a) expression was detected in Grade I, II and III STK, SCC and control samples of alveolar ridge keratoses (ARK). p16(INK4a) expression in STK and in ARK was typically weak but was relatively strong in all SCC. Strong p53 nuclear staining was detected in STK, SCC and ARK. HPV DNA was detected in Grade I, II and III STK, SCC and ARK, but did not correlate with p16(INK4a) expression. p16(INK4a) distribution did not correlate with STK grade and does not appear to be related to the detection of HPV DNA by PCR in either STK or in SCC. There is an apparent relationship between the grade of STK and the presence of HPV. HPV was rarely detected in high-grade lesions.

Potential biomarkers associated with discrimination between latent and active pulmonary tuberculosis.

SETTING: A third of the world's population has latent tuberculous infection (LTBI). Current TB diagnostics used in developing countries are ineffective and are unable to distinguish LTBI from active TB. Identifying biomarkers that could aid in the early detection of TB and in distinguishing TB states could be a major breakthrough in global TB control. OBJECTIVE: To identify potential immune biomarkers to distinguish active TB from LTBI. DESIGN: A cross-sectional study was conducted among 19 active TB patients, 8 TB-negative individuals (controls) and 16 LTBI non-human immunodeficiency virus infected individuals in Nairobi, Kenya. Excess supernatants from the QuantiFERON®-TB Gold In-Tube test were used to measure immune analytes using a Th17-focused Milliplex® assay. RESULTS: Overall antigen-specific responses were higher in the LTBI group than in active TB patients and controls. Interleukin (IL) 17F, macrophage inflammatory protein 3 alpha (MIP-3α), IL-13, IL-17A, IL-5, interferon-gamma (IFN-γ), IL-9, IL-1ß and IL-2 were significantly differentially produced by individuals with LTBI and active TB patients. Receiver operator curve analysis revealed good discriminative abilities of these analytes. Co-expression analysis highlighted uniquely co-expressed cytokine pairs between TB groups. CONCLUSION: These findings suggest that IL-17F, MIP-3α, IL-13, IL-17A, IL-5, IL-9, IL-1ß and IL-2, in addition to IFN-γ, could identify and uniquely discriminate between TB states.

Automation for clinical CD4 T-cell enumeration, a desirable tool in the hands of skilled operators.

BACKGROUND: Automation in HIV clinical flow cytometry when appropriately applied brings considerable standardisation benefits. The Canadian Immunology Quality Assessment Program (CIQAP) detected situations where operators did not manually override automated software in the event of improper output on the Epics XL and FC500 CD4 immunophenotyping platforms. The automated gating algorithm identifies lymphocytes using a double gate strategy based on CD45 × side scatter (SS) gating and a light scatter FS × SS gate known to fail with sub optimal specimens. METHOD: To generate correct interpretation and results CIQAP introduced a simple protocol modification, bypassing the light scatter gate to include all cells characterized by the CD45 gate. Seventeen problem cases were reanalysed for both absolute and relative T-cell subsets accuracy and compared to the CIQAP group mean values. Results were found to be associated with the percentage of lymphocytes excluded by the automated light scatter gate. RESULTS: The modified manual protocol resolved poor performance in 14 instances out of 17 problem cases. It was found to improve accuracy when the light scatter gate excluded greater than 5% of the cells. The remaining three cases had a lymphocyte recovery of greater than 94.6% in the original automated analysis. CONCLUSION: There is a risk in relying solely on automated gating procedures when using the Epics XL and FC500 CD4 immunophenotyping platforms. Laboratory managers have the responsibility to intervene when required. EQA providers are equally responsible to alert the clinical laboratories of the need to update operator training to deal with stressed specimens. © 2016 International Clinical Cytometry Society.

Comparison of methods for auto-coding causation of injury narratives.

Manually reading free-text narratives in large databases to identify the cause of an injury can be very time consuming and recently, there has been much work in automating this process. In particular, the variations of the naïve Bayes model have been used to successfully auto-code free text narratives describing the event/exposure leading to the injury of a workers' compensation claim. This paper compares the naïve Bayes model with an alternative logistic model and found that this new model outperformed the naïve Bayesian model. Further modest improvements were found through the addition of sequences of keywords in the models as opposed to consideration of only single keywords. The programs and weights used in this paper are available upon request to researchers without a training set wishing to automatically assign event codes to large data-sets of text narratives. The utility of sharing this program was tested on an outside set of injury narratives provided by the Bureau of Labor Statistics with promising results.

Food accessibility and food choice. A test of Schachter's externality hypothesis.

A set of naturalistic observations was conducted to examine Schachter's theory that obese individuals are more responsive to external food cues than persons of normal weight. During six days of observation at a large hospital cafeteria, experimenters manipulated the accessibility of high- and low-calorie desserts. No differences in selection by obese, overweight, and normal-weight individuals of meals or desserts were observed. All weight groups were equally responsive to the experimental manipulation of food cues.

Obesity and eating style.

Eating styles of 30 obese women and 37 matched women of normal weight were observed unobtrusively in a fast-food restaurant in the search for an "obese eating style" or other differences between the two groups. The size and character of the food was carefully matched by giving each woman a coupon entitling her to a free meal of either 985 or 1,800 calories. There were only small and inconsistent differences between obese and normal-weight women. No evidence of an "obese eating style" was found.

Addicted and nonaddicted drug users. A comparison of drug usage patterns.

Adetailed analysis of drug usage patterns of 101 multidrug-using soldiers disclosed little differentiation between individuals with a history of heroin addiction (N=68) from demographically similar multidrug-using (but nonaddicted) peers (N=33). Prior to the onset of herion addiction, relatively few differences in drug usage patterns emerged between the two groups, and what differences there were indicated more extensive drug use and a more rapid progression of drug use among the nonaddicted users. A "stepping-stone hypothesis" of heroin addiction is refuted by these data. Differences in drug use emerge after the intiation of heroin. Within a few months after first heroin use, those who did not become addicted returned to their preheroin experimentation levels of drug taking while the addicted group maintained an escalating pattern of opiate use.

IL-4 and IL-13 have overlapping but distinct effects on HIV production in monocytes.

In HIV-1-infected monocytes and monocytoid cell lines, viral expression can be observed as high-level production, restricted (chronic low-level) expression, and latency (no viral expression). Interleukin-13 (IL-13) and IL-4, which have remarkedly similar deactivating effects on inflammatory monocyte functions, were studied for their regulation of HIV expression in monocytes. Pretreatment of peripheral monocytes for 48-72 h with IL-13 markedly decreased acute HIV infection, whereas IL-4 increased it. Similar effects were seen when the U1 and R-THP-1 monocytoid cell lines with restricted HIV expression were treated with these cytokines. However, when these continuously producing cell lines were chronically treated with cytokines, IL-13 increased HIV production. Neither IL-4 nor IL-13 stimulated HIV expression in latently infected cells. In chronically infected cells, several cytokines reduced viral mRNA. Both IL-4 and IL-13 increased monocyte aggregate formation, but only IL-4 ultimately stimulated cytolysis of HIV-infected monocytes as well as increased apoptosis of U1. In the presence of tumor necrosis factor alpha or IL-6, which upregulate HIV expression, IL-13 could no longer suppress HIV expression. These results indicate that IL-4 and IL-13, although closely related in modulating monocyte function, can have divergent effects on HIV expression in monocytes. Collectively, these data suggest that there exists a complex cytokine tissue environment with positive regulators of HIV expression able to override negative regulators.

Parathyroid hypertension. A reversible disorder.

A retrospective study of 75 patients who were surgically cured of primary hyperparathyroidism from 1976 to 1984 was performed to evaluate the blood pressure and metabolic responses to parathyroid surgery. Published data on the population prevalence of hypertension (HT) in South Africa were used for comparison. The overall prevalence of HT before surgery was 47%, compared with 23% in the general population. Hypertension was most frequent in patients older than 60 years (62% vs 39% expected). Renal insufficiency was found in 13 of 35 hypertensive patients and in two of 40 normotensive patients. However, the prevalence of HT in patients with normal creatinine levels (37%) exceeded that expected. The frequency of urolithiasis and mean levels of serum and urine calcium and phosphate were similar in normotensive and hypertensive patients. Parathyroidectomy resulted in a substantial fall in both mean systolic and mean diastolic blood pressures in 54% of the hypertensive subjects, unrelated to improvement in renal function.

Chronic granulocytic leukemia in a patient with a renal allograft.

A man received a cadaver renal allograft for end-stage renal failure. After 35 months of immunosuppressive therapy with azathioprine and prednisone, he developed septicemia and a high leukocyte count. In spite of successful treatment of the infection, the leukocyte count continued to rise and a diagnosis of Philadelphia chromosome positive chronic granulocytic leukemia was made. An increased incidence of malignant disease, especially lymphoreticular malignancy, is well described in immunosuppressed patients with allografts. However, the association of chronic granulocytic leukemia and immunosuppressive therapy previously has not been reported. An additional etiological factor in this patient may have been the extensive diagnostic radiological investigations undertaken in childhood. The recent addition of allopurinol to the immunosuppressive therapy has normalized the platelet and leukocyte counts, probably by potentiating mercaptopurine.

Significance, definition, classification and risk factors of chronic kidney disease in South Africa.

Renal dysfunction or chronic kidney disease (CKD) is found in 10% of the global population and is classified into five stages according to the estimated glomerular filtration rate (eGFR). No matter where a patient lives, estimation of the GFR is mandatory for decision-making and obtained by the simple measurement of a serum creatinine level. The objective of diagnosing CKD lies in its future prevention, early detection and proper treatment, which will prevent or delay functional deterioration. Primary hypertension (PH) occurs in 25% of South Africa (SA)s black population and is the putative cause of stage 5 CKD in 40 - 60% of these patients. Moreover, in this group, stage 5 CKD occurs at a relatively young age (35 - 45 years) compared with other population groups in whom stage 5 CKD resulting from PH usually occurs between 60 and 70 years of age. In the cohort study, PH has been found in 12 - 16% of black school learners (mean age 17 years) compared with 1.8 - 2% of other ethnic groups (mixed race, Asian, white). End-stage renal failure (ESRF) is the fifth most common cause of death in SA, excluding post-traumatic cases. In addition, undiagnosed or poorly controlled PH is a potent risk factor for other cardiovascular disease (CVD), e.g. congestive cardiac failure, myocardial infarction, stroke. Significant protein is also associated with CVD and protein >1 g/d is a significant risk factor for ESRF.