Microneedle-devices provide a promising alternative to syringe-injection-based administration of chemotherapeutics. Dissolvable polymeric microneedles provide possibility of carrying greater payload and dual drugs. Here, we report development of polyvinyl pyrrolidone and polyvinyl alcohol composite dissolvable polymeric microneedle system for co-delivery of doxorubicin HCl and docetaxel. Microneedle patches were characterized using stereomicroscope, scanning electron microscope, texture analyzer and confocal microscope. The greatest amount of doxorubicin and docetaxel loaded within one microneedle patch was 533 ± 65 and 227 ± 23 µg, respectively. Ex-vivo studies in excised murine skin revealed insertion of microneedles and permeation of chemotherapeutics without lag time. Microneedles dissolved within 1 h of insertion in excised skin. Effectiveness of the delivery system was determined in 4T1 breast cancer cells xenografted athymic Balb/c mouse model. Intra-tumoral injection of doxorubicin and doxorubicin + docetaxel combination showed significant toxicity to animals evidenced by drastic reduction in the body weight and 100 percent death within 9-days and after 2-dose administration. Interestingly, doxorubicin and docetaxel administered using microneedles either alone or in combination showed significantly greater survival (100% survival after 16-days and 4-dose administration) compared with intratumoral injections. The normalized body weight, tumor volume and DNA fragmentation assay indicated superior effect of microneedle patch application. Furthermore, co-delivery of doxorubicin and docetaxel, controlled the tumor growth better than the administration of single molecules. Taken together, minimally invasive dissolvable microneedle patch application could compliment painful catheter assisted syringe injections to deliver combination chemotherapeutics.

Transdermal uptake models compliment in vitro and in vivo experiments in assessing risk of environmental exposures to semivolatile organic compounds (SVOCs). A key parameter for mechanistic models is the chemical driving force for mass transfer from environmental media to human skin. In this research, we measure this driving force in the form of fugacity for chemicals in cosmetic cream and use it to model uptake from cosmetics as a surrogate for condensed environmental media. A simple cosmetic cream, containing no target analytes, was mixed with diethyl phthalate (DEP), di-n-butyl phthalate (DnBP), and butyl paraben (BP) and diluted to make creams with concentrations ranging from 0.025% to 6%. The fugacity, relative to the pure compound, was measured using solid-phase micro extraction (SPME). We found that the relationship between the concentration and fugacity is highly nonlinear. The relative fugacity of the chemicals for a 2% w/w formulation was used in a diffusion-based model to predict transdermal uptake of each chemical and was compared with excretion data from a prior human subject study with the same formulation. Dynamic simulations of excretion are generally consistent with the results of the human subject experiment but sensitive to the input parameters, especially the time between cream application and showering.

While kidney donations stagnate, the number of people in need of kidney transplants continues to grow. Although transplanting culture-grown organs is years away, pursuing the engineering of the kidney de novo is a valid means of closing the gap between the supply and demand of kidneys for transplantation. The structural organization of a mouse kidney is similar to that of humans. Therefore, mice have traditionally served as the primary model system for the study of kidney development. The mouse is an ideal model organism for understanding the complexity of the human kidney. Nonetheless, the elaborate structure of the mammalian kidney makes the discovery of new therapies based on de novo engineered kidneys more challenging. In contrast to mammals, amphibians have a kidney that is anatomically less complex and develops faster. Given that analogous genetic networks regulate the development of mammalian and amphibian nephric organs, using embryonic kidneys of Xenopus laevis (African clawed frog) to analyze inductive cell signaling events and morphogenesis has many advantages. Pioneering work that led to the ability to generate kidney organoids from embryonic cells was carried out in Xenopus. In this review, we discuss how Xenopus can be utilized to compliment the work performed in mammalian systems to understand kidney development.

Aneuploidy, defined as structural and numerical aberrations of chromosomes, continues to draw attention as an informative effect biomarker for carcinogens and male reproductive toxicants. It has been well documented that aneuploidy is a hallmark of cancer. Aneuploidies in oocytes and spermatozoa contribute to infertility, pregnancy loss and a number of congenital abnormalities, and sperm aneuploidy is associated with testicular cancer. It is striking that several carcinogens induce aneuploidy in somatic cells, and also adversely affect the chromosome compliment of germ cells. In this paper we review 1) the contributions of aneuploidy to cancer, infertility, and developmental abnormalities; 2) techniques for assessing aneuploidy in precancerous and malignant lesions and in sperm; and 3) the utility of aneuploidy as a biomarker for integrated chemical assessments of carcinogenicity, and reproductive and developmental toxicity.

Larval zebrafish (Danio rerio) are a tool for assessing endocrine disruption during early development. Here, we investigated the extent to which a simple light/dark behavioral test at five days post fertilization could compliment current methods within the field. We exposed fertilized embryos to hormones (17β-estradiol, testosterone, dihydrotestosterone, 11-ketotestosterone, thyroxine, triiodothyronine, progesterone, and hydrocortisone) and other relevant compounds (17α ethinylestradiol, bisphenol A, bisphenol S, nonylphenol, flutamide, nilutamide, linuron, drospirenone, potassium perchlorate, mifepristone, and fadrozole) to screen for behavioral effects between 96 and 118h post fertilization (hpf). With the exception of progesterone, all the hormones tested resulted in altered behaviors. However, some inconsistencies were observed regarding the age of the larvae at testing. For example, the xenoestrogens 17α- ethinylestradiol and nonylphenol had behavioral effects at 96hpf, but not at 118hpf. Furthermore, although thyroxine exposure had pronounced effects on behavior, the thyroid disruptor potassium perchlorate did not. Finally, we were unable to demonstrate a role of nuclear receptors following testosterone and 17α- ethinylestradiol exposure, as neither the androgen receptor antagonist flutamide nor the general estrogen receptor inhibitor fulvestrant (ICI) could rescue the observed behavioral effects, respectively. Similarly, molecular markers for androgen and estrogen disruption were upregulated at concentrations below which behavioral effects were observed. These results demonstrate hormones and endocrine disruptors can alter the behavior of larval zebrafish, but the mechanistic pathways remain unclear.

In July 2018, the Food and Drug Administration warned about a possible relationship between dilated cardiomyopathy (DCM) in dogs and the consumption of dog food formulated with potatoes and pulse ingredients. This issue may impede utilization of pulse ingredients in dog food or consideration of alternative proteins. Pulse ingredients have been used in the pet food industry for over 2 decades and represent a valuable source of protein to compliment animal-based ingredients. Moreover, individual ingredients used in commercial foods do not represent the final nutrient concentration of the complete diet. Thus, nutritionists formulating dog food must balance complementary ingredients to fulfill the animal's nutrient needs in the final diet. There are multiple factors that should be considered, including differences in nutrient digestibility and overall bioavailability, the fermentability and quantity of fiber, and interactions among food constituents that can increase the risk of DCM development. Taurine is a dispensable amino acid that has been linked to DCM in dogs. As such, adequate supply of taurine and/or precursors for taurine synthesis plays an important role in preventing DCM. However, requirements of amino acids in dogs are not well investigated and are presented in total dietary content basis which does not account for bioavailability or digestibility. Similarly, any nutrient (e.g., soluble and fermentable fiber) or physiological condition (e.g., size of the dog, sex, and age) that increases the requirement for taurine will also augment the possibility for DCM development. Dog food formulators should have a deep knowledge of processing methodologies and nutrient interactions beyond meeting the Association of American Feed Control Officials nutrient profiles and should not carelessly follow unsubstantiated market trends. Vegetable ingredients, including pulses, are nutritious and can be used in combination with complementary ingredients to meet the nutritional needs of the dog.

Urogenital schistosomiasis is a chronic parasitic disease that causes severe morbidity among schoolchildren in many poor-resource communities in Nigeria. We investigated the prevalence, intensity, and risk factors of the infection in three communities of Kwara State to ascertain the current status of the disease. Of the 724 urine samples screened, using filtration method, 332 (45.6%) school-aged children were infected with average intensity and mean population eggs load of 127.9 eggs/10 ml of urine and 0.794, respectively. Prevalence and intensity of infection varied with communities: high in Ajase-Ipo (57.1%; X = 100.7 ± 23.01 eggs/10 ml) and low in Shonga (37.5%; X = 91.4 ± 78.0). Infection was significantly (P < 0.05) higher in males (50.8%) than the females (42.4%). Similarly, infection significantly (P < 0.05) increased with increasing age. Multivariate logistic analysis of risk factors revealed that lack of portable drinking water (adjusted odd ratio (aOR) = 4.76; 95% CI = 2.64-5.98), unemployment (aOR = 2.23; 1.87-2.294), lack of knowledge of infection (aOR = 2.16; 0.59-3.83), and frequent contact with contaminated water bodies (aOR = 2.01; 1.45-2.70) were important predictors of urinary schistosomiasis. Therefore, continuous evaluation of the intervention strategies that address risk factors must compliment Mass Drug Administration to curtail the transmission and debilitating health consequences of infection in endemic settings.

DNA analysis is becoming routine in the clinical laboratory for the diagnosis of human diseases using various tissue sources. Most clinical specimens are followed by tracking forms that include patient demographic data, accession number, and date and time of collection. As part of a thorough quality assurance program, proper documentation of test requisitions and tracking forms is mandatory. Despite these efforts, specimen mislabeling or other mix-ups can, and do, occur. We demonstrate the utility of the PM + DQA1 typing kit and STR analysis using the Visible Genetics automated DNA sequencing system in the proper identification of such clinical specimens as urine, blood, and paraffin-embedded tissues. In each case, sufficient DNA was extracted from these specimen types using a nonorganic extraction protocol for typing purposes. We conclude that DNA typing methods are feasible for distinguishing clinical laboratory specimens of questionable identity and compliment existing quality assurance techniques.

UNASSIGNED

The way we view ourselves may play an important role in our responses to interpersonal interactions. In this study, we investigate how feedback valence, consistency of feedback with self-knowledge and global self-esteem influence affective and neural responses to social feedback.

UNASSIGNED

Participants (N = 46) with a high range of self-esteem levels performed the social feedback task in an MRI scanner. Negative, intermediate and positive feedback was provided, supposedly by another person based on a personal interview. Participants rated their mood and applicability of feedback to the self. Analyses on trial basis on neural and affective responses are used to incorporate applicability of individual feedback words.

UNASSIGNED

Lower self-esteem related to low mood especially after receiving non-applicable negative feedback. Higher self-esteem related to increased PCC and precuneus activation (i.e., self-referential processing) for applicable negative feedback. Lower self-esteem related to decreased mPFC, insula, ACC and PCC activation (i.e, self-referential processing) during positive feedback and decreased TPJ activation (i.e., other referential processing) for applicable positive feedback.

UNASSIGNED

Self-esteem and consistency of feedback with self-knowledge appear to guide our affective and neural responses to social feedback. This may be highly relevant for the interpersonal problems that individuals face with low self-esteem and negative self-views.

BACKGROUND

It has been over a decade since a comprehensive study has been published that has examined sarcoidosis deaths at the national level. The purpose of this study was to analyze sarcoidosis as the underlying cause of death using current national death certificate data. Results from this project can be used to evaluate and compare trends of sarcoidosis reported deaths across the U.S.

METHODS

Mortality data from 1999 to 2016 were provided by the National Vital Statistics System (NVSS) with sarcoidosis (ICD-D86.X) as the underlying cause of death from all resident death certificates filed in the 50 states and the District of Columbia (DC). Data were analyzed using CDC WONDER, a web-based public health database and analysis tool. Queries were used to generate number of deaths, along with unadjusted and age-adjusted death rates with 95% confidence intervals and standard errors for groups including year, census region, gender, age group, race/ethnicity and state. Joinpoint regression analysis was used to test the significance of trends in race and gender-specific rates for the 1999-2016 study period.

RESULTS

From 1999 to 2016, there were a total of 16,665 sarcoidosis reported deaths in the U.S. The overall age-adjusted mortality rate increased from 2.1 (deaths per 1,000,000) in 1999 to 3.1 in 2002, but then remained relatively stable thereafter until the end of the study period. Female deaths increased 32.0% (from 2.5 to 3.3 per 1,000,000), while male deaths increased 73.3% (from 1.5 to 2.6 deaths per 1,000,000). The highest age-adjusted death rates were among black females (17.0 deaths per 1,000,000), and black males (12.4 deaths per 1,000,000). At the regional level, the southern U.S. had the highest overall mean age-adjusted mortality rate (3.7 deaths per 1,000,000), while black females in the Midwest (18.7 per 1,000,000) had the highest race-specific reported death rate.

CONCLUSIONS

The detected increase in the total number of deaths and age-adjusted rates of sarcoidosis deaths in the U.S. is a serious health concern. Factors that contribute to sarcoidosis deaths remain uncertain and more epidemiological research studies are needed to compliment current bench science to explore and examine factors that contribute to this multifactorial, chronic disease.

The Comprehensive Test of Nonverbal Intelligence, or CTONI, has become an essential compliment to the traditional tests of intelligence, such as the WISC-III, DTLA-3, and the Binet. The CTONI provides examiners with a measure of nonverbal reasoning that requires no spoken language or complex motor skills. The CTONI has been proven to be unbiased with regard to gender, minority, or disabling condition. Finally, it is possible to estimate the intelligence of people without the contamination of social, ethnic, or disability bias.

The study explores how data collated from rapid assessment can enhance those produced by national level surveillance systems, in this case the national drug information (NADI) system in Malaysia. Qualitative data were collected in keeping with internationally accepted guidance on rapid assessment methods in the field of substance use. An inductive research strategy was employed. The rapid assessment produced multiple data on local drug use practices and how these were influenced by the contexts of use. The assessment points to the importance of collecting data not only on patterns of drug use but also on the health and social consequences of drug use. We suggest that the current national drug information system places greater emphasis on behavioural and health-related variables in order to better understand the potential relationships between drug use and health-related risk, including HIV/AIDS.

BACKGROUND

Rural experience for dental students can provide valuable clinical education, change attitudes to rural practice, and make a valuable contribution to clinical service provision. The aim of this paper is to assess the costs and benefits of service delivery by students through rural training programmes.

METHODS

Groups of two students worked in the public dental clinics in adjacent rural centres where there had been long-term difficulties in recruiting staff. The costs and benefits of the programme were assessed by the impact on waiting lists, the total cost per patient of a course of care and by the marginal cost of adding service provision by students to existing arrangements.

RESULTS

The total costs of emergency and complete treatment provided by students were greater than the costs of treatment provided by public-sector dentists but less than the costs of private providers treating public patients. However, the value of services were greater when care was provided by students or private providers and the marginal cost of students providing services was 50-70 per cent of the cost of care provided by public dentists.

CONCLUSIONS

This assessment suggests that the service benefits achieved compliment the primary objective of influencing the attitude of students to rural practice.

Increasing amounts of immune complexes (IC) accumulated in the blood of C57BL/6J (H-2b) mice infected for 16 weeks with alveolar hydatid cysts. The circulating IC were detected both quantitatively (precipitation with 3.75% polyethylene glycol) and qualitatively (Raji cell binding assay) in the sera of hydatid-mice. The levels of serum IC roughly parallel the increase in weight of the larval cyst masses (LCM) and were inversely reacted to serum compliment activity; the latter was markedly reduced at 14 weeks postinfection. The solubilized serum IC was reacted with appropriate antisera in gel precipitation test; it consisted of E. multilocularis antigens, IgGl, IgG2b, IgM and C3. Mouse IgG, IgM, complement and hydatid antigens were also detected by the indirect immunofluorescent technique in kidney sections of hydatid-mice infected 14 weeks before. The role of IC is discussed with reference to immunodepression and growth of the LCM in hydatid mice.

Intimate partner violence (IPV) is a pervasive social concern that may be exacerbated by high rates of alcohol dependence among perpetrators. Society has attempted to combat IPV through various legal interventions, but the effects of specific legal factors on behavioral change and treatment compliance remain largely unexamined. The primary focus of the current study was to comprehensively evaluate the impact of various legal factors (i.e., judicial mandate, judicial monitoring, stage of change, and stake in conformity) on mandatory treatment compliance and behavioral change over a 12 week post-adjudication period among a high-risk sample of alcohol dependent IPV offenders (N = 60). Growth curve analyses revealed effects of judicial monitoring and stage of change such that participants reporting low perceived judicial monitoring and early stages of change reported higher initial levels and a more rapid reduction in IPV than those reporting high perceived judicial monitoring and late stages of change, who reported consistently low IPV. Although we found that legal factors were poor predictors of treatment compliance and alcohol use during treatment, the association between alcohol and IPV was moderated by the legal factors. Stake in conformity was negatively associated with IPV among low alcohol users and positively associated among high alcohol users whereas stage of change was negatively associated with IPV among high alcohol users. The current results suggest that pretreatment legal factors may represent an important consideration in reducing IPV among alcohol dependent offenders. Further research is required to determine the efficacy of legal factors in isolation of treatment as well as methods of manipulating these factors to optimally compliment a prescribed course of treatment.

Neurosurgeons are often faced with a lesion that is obvious on magnetic resonance imaging (MRI), but not well-defined on computerized tomography (CT). However, questions remain regarding the distortion inherent in MRI images. Therefore the following comparative study was conducted. Five patients with intracranial lesions (1 lymphoma, 1 multiple sclerosis, 2 glioblastomas, and 1 AVM), underwent both CT and MRI for purpose of stereotactic biopsy or radiosurgery. The Brown-Roberts-Wells CT and MRI compatible localizing rings were used. Coordinates of the left optic nerve-globe junction, the aqueduct, pineal and optic chiasm were recorded from both CT and MRI. With MRI, all three imaging planes, axial, coronal and sagittal, were used. Coordinates were calculated in millimeters and submitted to statistical analysis using Pierson correlation coefficients. In all, there were 17 CT, 17 MRI axial, 13 MRI coronal, and 13 MRI sagittal coordinates. The analysis revealed that of the MRI coordinates, the axial coordinates were more available and retrievable. Lateral targets, such as the left optic nerve-globe junction, were more difficult to identify on coronal and sagittal images and often lacked a full compliment of fiducial points for calculation. The correlation with CT was best for axial, followed by coronal and then sagittal MRI planes. Geometric image distortion occurs owing to nonlinearity of the magnetic field as well as magnet susceptibility to different tissues at an interface. We feel that MRI stereotaxy can be utilized in conjunction with CT for verification of lesion coordinates in relatively large lesions, and particularly in those not well delineated on CT.

Research investigating 'mirror neurons' has demonstrated the presence of an observation-execution matching system in humans. One hypothesized role for this system might be to aid in action understanding by encoding the underlying intentions of the actor. To investigate this hypothesis, we asked participants to observe photographs of an actor making orofacial gestures (implying verbal or non-verbal acts), and to produce syllables that were compatible or incompatible with the gesture they observed. We predicted that if mirror neurons encode the intentions of an actor, then the pictures implying verbal gestures would affect speech production, whereas the non-verbal gestures would not. Our results showed that the observation of compatible verbal gestures facilitated verbal responses, while incompatible verbal gestures caused interference. Although this compatibility effect did not reach statistical significance when the photographs implied a non-verbal act, responses were faster on average when the gesture implied the use of similar articulators as those involved with the production of the target syllable. Altogether, these behavioral findings compliment previous neuroimaging studies indicating that static pictures portraying gestures activate brain regions associated with an observation-execution matching system.

Two image reconstruction methods currently dominate parallel MR imaging: SENSE and GRAPPA. While both seek to reconstruct images from subsampled multi-channel MRI data, there exist fundamental differences between the two. In particular, SENSE reconstructs an image of the excited spin-density directly whereas GRAPPA reconstructs estimates of the fully sampled raw coil data and then combines them to obtain an image. In this work we show that these differences can be exploited such that each method can compliment the other. In the case of SENSE, which requires an estimate of the coil sensitivity map before reconstruction, one can use GRAPPA to improve the coil sensitivity estimates. Alternatively, using coil sensitivity estimates and the SENSE reconstruction equations, one can improve the GRAPPA reconstruction parameter estimation. Together, these approaches can provide higher image quality than either method alone.

Online fitness communities are a recent phenomenon experiencing growing user bases. They can be considered as online social networks in which recording, monitoring, and sharing of physical activity (PA) are the most prevalent practices. They have added a new dimension to the social experience of PA in which online peers function as virtual PA partners or supporters. However, research into seeking and receiving computer-mediated social support for PA is scarce. Our aim was to study to what extent using online fitness communities and sharing physical activities with online social networks results in receiving various types of online social support.

Two databases, one containing physical activities logged with Strava and one containing physical activities logged with RunKeeper and shared on Twitter, were investigated for occurrence and type of social support, by means of a deductive content analysis.

Results indicate that social support delivered through Twitter is not particularly extensive. On Strava, social support is significantly more prevalent. Especially esteem support, expressed as compliments for the accomplishment of an activity, is provided on both Strava and Twitter.

The results demonstrate that social media have potential as a platform used for providing social support for PA, but differences among various social network sites can be substantial. Especially esteem support can be expected, in contrast to online health communities, where information support is more common.

The Scale for Interpersonal Behaviour (SIB), a multidimensional, self-report measure of state assertiveness, was administered to a nationwide sample of 2375 undergraduates enrolled at 11 colleges and universities across the USA. The SIB was developed in the Netherlands for the independent assessment of both distress associated with self-assertion in a variety of social situations and the likelihood of engaging in a specific assertive response. This is done with four factorially-derived, first-order dimensions: (i) Display of negative feelings (Negative assertion); (ii) Expression of and dealing with personal limitations; (iii) Initiating assertiveness; and (iv) Praising others and the ability to deal with compliments/praise of others (Positive assertion). The present study was designed to determine the cross-national invariance of the original Dutch factors and the construct validity of the corresponding dimensions. It also set out to develop norms for a nationwide sample of US students. The results provide further support for the reliability, factorial and construct validity of the SIB. Compared to their Dutch equivalents, US students had meaningfully higher distress in assertiveness scores on all SIB scales (medium to large effect sizes), whereas differences on the performance scales reflected small effect sizes. The cross-national differences in distress scores were hypothesized to have originated from the American culture being more socially demanding with respect to interpersonal competence than the Dutch, and from the perceived threats and related cognitive appraisals that are associated with such demands.

OBJECTIVE

To determine the clinical usefulness of computed tomography (CT) angiography in the evaluation of cerebral aneurysms.

METHODS

From October 1994 through April 1996, 26 patients with 30 surgical proven intracranial aneurysms underwent both CT angiography and catheter cerebral angiography. The findings of the two methods were reviewed independently and then compared with each other.

RESULTS

Comparing with catheter angiography, CT angiography was superior in demonstrating the aneurysmal neck in seven aneurysms but was inferior in one. The thrombosed part and calcification of aneurysms were clearly demonstrated on CT angiograms. CT angiography also aided in differentiating tight vascular loops from aneurysms. On CT angiograms, one posterior communicating arterial aneurysm was overlooked and another anterior choroidal artery aneurysm was misinterpreted as a posterior communicating artery aneurysm. Of note were two patients in whom the infundibulum of the orbitofrontal artery was misinterpreted as the anterior communicating artery aneurysm.

CONCLUSIONS

CT angiography can compliment conventional catheter angiography for its better demonstration of the 3-dimensional anatomy. It can provide surgical information about the neck, calcification and thrombosed part of an aneurysm and its relation to adjacent structures. However, caution is advocated because CT angiography may fail to demonstrate small but important vessels such as posterior communicating, anterior choroidal and orbitofrontal arteries. Recognition of the limitations of CT angiography is important in minimizing interpretation errors.

Complaints against nurses can be made on several grounds and orders, including removal from the registry of nurses, can be made as a result of these complaints. Boundary violations generally relate to complaints around criminal charges, unsatisfactory professional conduct or professional misconduct or a lack of good character. This article explores the spectrum of boundary violations in the nurse-patient relationship by reviewing disciplinary cases from the New South Wales Nurses and Midwives Tribunal and Professional Standards Committees. The complaints spanned a spectrum of behaviours, from minor infringements such as inappropriate compliments to intimate touching and sexual intercourse. Furthermore, the majority of respondents were men, although men comprise a minority of the nursing profession. This phenomenon is discussed in terms of gender stereotyping and nursing work. In addition, the possibility that improved supervision may have gone some way to preventing the violations is explored.

BACKGROUND

The devastating viral disease of small ruminants namely Peste des petits ruminants (PPR) declared as target for "Global Eradication" in 2015 by the Food and Agriculture Organization (FAO) and the World Organization for Animal Health (OIE). For a successful eradication campaign, molecular diagnostic tools are preferred for their specificity, efficacy and robustness to compliment prophylactic measures and surveillance methods. However, molecular tools have a few limitations including, costly equipment, multi-step template preparation protocols, target amplification and analysis that restrict their use to the sophisticated laboratory settings. As reverse transcription-loop mediated isothermal amplification assay (RT-LAMP) has such an intrinsic potential for point of care diagnosis, this study focused on the genetic detection of causative PPR virus (PPRV) in field conditions. It involves the use of a sample buffer that can precipitate out virus envelope and capsid proteins through ammonium sulphate precipitation and exposes viral RNA, present in the clinical sample, to the LAMP reaction mixture.

RESULTS

The test was evaluated using 11 PPRV cultures, and a total of 46 nasal swabs (n = 32 collected in the field outbreaks, n = 14 collected from experimentally inoculated animals). The RT-LAMP was compared with the reverse transcription-PCR (RT-PCR) and real-time quantitative RT-PCR (RT-qPCR) for its relative specificity, sensitivity and robustness. RT-LAMP detected PPRV in all PPRV cultures in or less than 30 min. Its detection limit was of 0.0001TCID50 (tissue culture infective dose-50) per ml with 10-fold higher sensitivity than that of RT-PCR. In 59.4% of the field samples, RT-LAMP detected PPRV within 35-55 min. The analytical sensitivity and specificity of the RT-LAMP were equivalent to that of the RT-qPCR. The time of detection of PPRV decreased by at least forty minutes or 3-4 h in case of in the RT-LAMP as compared with the RT-qPCR and the RT-PCR, respectively.

CONCLUSIONS

The sensitive and specific RT-LAMP test developed in this study targeting a small fragment of the N gene of PPRV is a rapid, reliable and applicable molecular diagnostic test of choice under the field conditions. RT-LAMP requiring minimal training offers a very useful tool for PPR diagnosis especially during the "Global PPR Eradication Campaign".