Recovery of Recombinant Canine Distemper Virus That Expresses CPV-2a VP2: Uncovering the Mutation Profile of Recombinant Undergoing 50 Serial Passages In Vitro

Canine distemper and canine parvoviral enteritis are infections caused by the canine distemper virus (CDV) and canine parvovirus type 2 (CPV-2), respectively. They are two common infectious diseases that cause high morbidity and mortality in affected dogs. Combination vaccines have been broadly used to protect dogs from infections of CDV, CPV-2, and other viruses. VP2 is the most abundant protein of the CPV-2 capsid. It elicits potent immunity in animals and, therefore, is widely used for designing subunit antigen-based vaccines. In this study, we rescued a recombinant CDV (QN vaccine strain) using reverse genetics. The recombinant CDV (rCDV-VP2) was demonstrated to express stably the VP2 in cells for at least 33 serial passages in vitro. Unfortunately, a nonsense mutation was initially identified in the VP2 open reading frame (ORF) at passage-34 (P34) and gradually became predominant in rCDV-VP2 quasispecies with passaging. Neither test strip detection nor indirect immunofluorescence assay demonstrated the expression of the VP2 at P50. The P50 rCDV-VP2 was subjected to next-generation sequencing, which totally identified 17 single-nucleotide variations (SNVs), consisting of 11 transitions and 6 transversions. Out of the 17 SNVs, 1 and 9 were identified as nonsense and missense mutations, respectively. Since the nonsense mutation arose in the VP2 ORF as early as P34, an earlier rCDV-VP2 progeny should be selected for the vaccination of animals in future experiments.

EFFECT OF MENSTRUATION ON TEAR PRODUCTION AMONG UNDERGRADUATE STUDENTS OF MADONNA UNIVERSITY, ELELE CAMPUS, RIVERS STATE NIGERIA

Purpose: The menstrual cycle has been reported to influence many other physiological processes in women. This study ascertained the effect of menstruation on oculovisual status with emphasis on tear production among young female undergraduates of Madonna University, Elele Campus Rivers State Nigeria. Methodology: The study involved 103 female undergraduates with fairly regular menstrual cycle selected through convenience sampling technique. Their age ranged from 16-30 years with a mean age of 20.82 ± 3.205 years and a median age of 20 years. A well-structured interviewer administered questionnaire was used to elicit oculovisual symptoms associated with the menstrual cycle. Schirmer’s test strip was used to measure tear volume at pre-determined intervals of the menstrual flow period. External and internal eye examinations were also carried out using the Pen torch and ophthalmoscope respectively. Data collected were analysed using the Statistical Package for Social Sciences (SPSS) version 21 and results were expressed using tables and correlation of variables expressed using Dunnett’s multiple comparison test. Findings: There was a significant increase (p ˂ 0.05) in tear production during the first two days of the flow and a slight decrease during the last two days. Comparison of the right and left eyes showed no statistically significant (p ˃ 0.05) difference. Variation in tear production associated with menstruation with respect to age showed statistically significant (p ˂ 0.05) differences for all age groups except for the right eye of subjects within age group 26–30 years. While 52.4% (n=54 ) of the ladies have no oculovisual complaint, the most common oculovisual complaint was headache 42.7% (n = 44) followed by blurry vision 2.91 % (n = 3) and excessive tearing 1.94% (n = 2). Contribution to Knowledge: The study revealed that physiological changes likely resulting from hormonal variation during the menstrual flow influences tear volume and may cause visual discomfort among women within the reproductive age and recommended the need for tear volume assessment and detailed case history prior to management of complaints associated with dry eye and dispensing of contact lenses for pre and post-menopausal women.

Novel Latex Microsphere Immunochromatographic Assay for Rapid Detection of Cadmium Ion in Asparagus

Recently, concerns about heavy metal cadmium ion (Cd2+) residue in asparagus have been frequently reported, and there is an urgent need to develop an effective, sensitive, and rapid detection method for Cd2+. In this study, we innovatively combined molecular microbiology to carry out the comparative screening of Cd2+ chelators in a green, efficient, and specific way. The knock-out putative copper-transporter gene (pca1Δ) yeast strain with high sensitivity to Cd2+ was first used to screen the Cd2+ chelator, and the optimum chelator 1-(4-Isothiocyanatobenzyl)ethylenediamine-N,N,N,N′-tetraacetic acid (ITCBE) was obtained. Additionally, a rapid latex microsphere immunochromatographic assay (LMIA) was developed, based on the obtained monoclonal antibody (mAb) with high specificity and high affinity (affinity constant Ka = 1.83 × 1010 L/mol), to detect Cd2+ in asparagus. The 50% inhibitive concentration (IC50) of test strip was measured to be 0.2 ng/mL, and the limit of detection (IC10) for qualitative (LOD, for visual observation) and quantitative detection (LOQ, for data simulation) of the test strip was 2 ng/mL and 0.054 ng/mL, respectively. In all, the developed mAb-based LMIA shows a great potential for monitoring Cd2+ in asparagus, even in vegetable samples.

Application of the optoacoustic effect to measure glucose concentration

Currently, there are only two invasive methods available for measuring blood glucose: the first, the most common method, requires blood to be drawn, which is then transported and analyzed to a clinical laboratory, applying the blood to a glucometer test strip. Sampling blood, especially in children, is a rather complicated process, in addition, the results of clinical trials take time. Therefore, non-invasive, continuous monitoring of glucose by optoacoustic method is quite promising. We used an optoacoustic method with a high spatial resolution, which makes it possible to measure the concentration of glucose in the blood. Experimental measurements were carried out in porcine blood at various glucose concentrations

Clinical Trial on the Usefulness of On-Site Evaluation of Canine Fetal Fluids by Reagent Test Strip in Puppies at Elective Caesarean Section

The reagent urinary test strips (TS) marketed for urines represent the first-line diagnostic tool in many instances. Therefore, the aim of the study was to assess the usefulness of TS for the on-field evaluation of fetal fluids’ composition in newborn dogs at elective caesarean section. Of a total of 137 puppies born at term, 127 survived and 10 did not survive. One hundred and thirteen amniotic and 107 allantoic samples from surviving newborns were collected, and 8 amniotic and allantoic fluids were collected from the non-surviving newborns and assessed by strips. Significantly lower amounts of amniotic glucose and higher amounts of amniotic and allantoic nitrites, amniotic protein, allantoic urobilinogen, and amniotic bilirubin concentrations were found in non-surviving when compared to surviving newborns. In the surviving ones, higher specific gravity and bilirubin concentrations, and lower pH, were found in allantoic than in amniotic fluids. Higher amniotic and allantoic glucose concentrations, higher amniotic and allantoic pH, and lower amniotic and allantoic protein concentrations were found in medium/large- than in small-sized puppies. The TS allowed the quick evaluation of fetal fluids in puppies at birth. The differences between surviving and non-surviving puppies seem to suggest that the on-site analysis of fetal fluids’ composition by TS could represent a first-line diagnostic tool in the field of canine neonatology, allowing the quick recognition of puppies needing assistance as a complementary tool for clinical evaluation.

Desain Alat Ukur Kadar Asam Urat Non-Invasive Dengan Sensor Near Infrared

Gout disease or commonly known as gout arthritis is a disease caused by the accumulation of monosodium urate crystals in the body. Monitoring blood uric acid levels is currently still using invasive techniques by taking blood samples to be included in the test strip, this invasive technique measurement requires money and time to come to health clinics so that many people cannot monitor blood uric acid levels on a regular basis. This study aims to develop a measuring instrument for monitoring blood uric acid levels using sensors by utilizing the absorption and reflectance of infrared rays so that gout sufferers and other individuals can monitor blood uric acid levels regularly and are easy to use. The method of developing non-invasive techniques focuses on monitoring blood uric acid levels using a NIR sensor with an IR LED with a wavelength of 940 nm as a transmitter and a photodiode as a detector at a wavelength of 700-1300 nm and a microcontroller as a minimum system for control. The first stage is measuring uric acid levels with invasive techniques on participants and at the same time measuring voltages with sensors, the results with invasive techniques are correlated with sensor output voltages to obtain non-linear equations in polynomial form, for conversion programs on the microcontroller. The second stage is measuring uric acid levels with invasive techniques and invasive measurements on participants at the same time. Both monitoring results were analyzed by simple ANOVA statistics and calculated SEP and RMSE to determine the accuracy of the prediction equation and its accuracy value.