Rapid Detection of Porcine DNA in Meatball Using Recombinase Polymerase Amplification Couple with Lateral Flow Immunoassay for Halal Authentication.

Point-of-care diagnostic methods for animal species determination are critical for rapid, simple, and accurate enforcement of food labelling. PCR is the most common method for species identification. However, the requirement of using a thermal cycler created drawbacks for the PCR application, particularly in low-resource settings. Hence, in this study, a method for porcine DNA detection using recombinase polymerase amplification (RPA), coupled with nucleic acid lateral flow immunoassay (NALFIA), was developed. Porcine-specific primers targeting pig (Sus scrofa) cytochrome b gene fragments specifically amplify a 197 bp fragment of the mitochondrial gene as being visualized by 2% agarose gel and PCRD NALFIA. The reaction temperature and time were 39 °C and 20 min, respectively. Herein, the specificity of the primers to porcine was confirmed after being assayed against six animal species, namely cow, goat, chicken, duck, dog, and rabbit. The porcine-specific RPA assay shows a high limit of detection of 0.01 ng/µL pork DNA. Based on the preliminary performance data obtained from this study, the potential of this method as a rapid and sensitive tool for porcine DNA detection in meat-based products is foreseen.

Automated diabetic retinopathy screening for primary care settings using deep learning.

Diabetic Retinopathy (DR) is one of the leading causes of blindness in the United States and other high-income countries. Early detection is key to prevention, which could be achieved effectively with a fully automated screening tool performing well on clinically relevant measures in primary care settings. We have built an artificial intelligence-based tool on a cloud-based platform for large-scale screening of DR as referable or non-referable. In this paper, we aim to validate this tool built using deep learning based techniques. The cloud-based screening model was developed and tested using deep learning techniques with 88702 images from the Kaggle dataset and externally validated using 1748 high-resolution images of the retina (or fundus images) from the Messidor-2 dataset. For validation in the primary care settings, 264 images were taken prospectively from two diabetes clinics in Queens, New York. The images were uploaded to the cloud-based software for testing the automated system as compared to expert ophthalmologists' evaluations of referable DR. Measures used were area under the curve (AUC), sensitivity, and specificity of the screening model with respect to professional graders. The screening system achieved a high sensitivity of 99.21% and a specificity of 97.59% on the Kaggle test dataset with an AUC of 0.9992. The system was also externally validated in Messidor-2, where it achieved a sensitivity of 97.63% and a specificity of 99.49% (AUC, 0.9985). On primary care data, the sensitivity was 92.3% overall (12/13 referable images are correctly identified), and overall specificity was 94.8% (233/251 non-referable images). The proposed DR screening tool achieves state-of-the-art performance among the publicly available datasets: Kaggle and Messidor-2 to the best of our knowledge. The performance on various clinically relevant measures demonstrates that the tool is suitable for screening and early diagnosis of DR in primary care settings.

Determination of potential oxidative damage, hepatotoxicity, and cytogenotoxicity in male Wistar rats: Role of indomethacin.

The present study demonstrated the indomethacin (INDO) induced oxidative stress, hepatotoxicity, and genotoxicity in male Wistar rats. Animals were orally administrated INDO at doses of 0.302 and 0.605 (mg/kg b.w.) for 2 weeks. Reduced glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), and lipid peroxidation (LPO) activities/levels were measured in the liver, kidney, and brain tissues. The aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP) activities, total bilirubin (TBIL) levels, and histopathological changes were determined in the liver tissues. Micronucleus frequency (micronucleus test) and DNA damage (comet assay) tests were performed in the bone marrow cells and leukocytes, respectively. Results show that INDO treatment decreased the GSH, SOD, and CAT levels/activities and increased the LPO, ALT, AST, ALP, and TBIL activities/levels. INDO induced significant hepatic injury and micronucleus and DNA damage. Thus, the current investigations confirm the oxidative stress, hepatotoxic, and genotoxic properties of INDO in the male Wistar rats.

Motor Nerve Conduction Velocity In Postmenopausal Women with Peripheral Neuropathy.

INTRODUCTION: The post-menopausal phase is characterized by a decline in the serum oestrogen and progesterone levels. This phase is also associated with higher incidence of peripheral neuropathy. AIM: To explore the relationship between the peripheral motor nerve status and serum oestrogen and progesterone levels through assessment of Motor Nerve Conduction Velocity (MNCV) in post-menopausal women with peripheral neuropathy. MATERIALS AND METHODS: This cross-sectional study was conducted at Jawaharlal Nehru Medical College during 2011-2013. The study included 30 post-menopausal women with peripheral neuropathy (age: 51.4±7.9) and 30 post-menopausal women without peripheral neuropathy (control) (age: 52.5±4.9). They were compared for MNCV in median, ulnar and common peroneal nerves and serum levels of oestrogen and progesterone estimated through enzyme immunoassays. To study the relationship between hormone levels and MNCV, a stepwise linear regression analysis was done. RESULTS: The post-menopausal women with peripheral neuropathy had significantly lower MNCV and serum oestrogen and progesterone levels as compared to control subjects. Stepwise linear regression analysis showed oestrogen with main effect on MNCV. CONCLUSION: The findings of the present study suggest that while the post-menopausal age group is at a greater risk of peripheral neuropathy, it is the decline in the serum estrogen levels which is critical in the development of peripheral neuropathy.

The role of the endogenous antioxidant enzymes and malondialdehyde in essential hypertension.

CONTEXT: Oxidative Stress is caused by an imbalance between the production of reactive oxygen species and a biological system's ability to readily detoxify the reactive intermediates. AIMS: 1. To compare the levels of Malondialdehyde (MDA), in hypertensive and normotensive subjects. 2. To compare the levels of the antioxidant enzymes, namely, Catalase, Glutathione peroxidase (GPX) and Superoxide Dismutase (SOD) in hypertensive and normotensive subjects. 3. To determine the correlation between the MDA levels and the mean arterial pressure (MAP) among hypertensive subjects. 4. To determine the correlation between the antioxidant enzyme levels and MAP among the hypertensive subjects and to evaluate the effect of 6 months of antihypertensive therapy with a tight blood pressure control on the MDA levels. Materials and Methods : In this cross sectional study, 25 normotensive and 40 hypertensive subjects were recruited. The hypertensive subjects were further subdivided into three subgroups: Prehypertensives, Stage I hypertensives and Stage II hypertensives. All the subjects underwent a blood pressure measurement and the markers of oxidative stress in their sera were estimated. The subjects of Stage I hypertension and Stage II hypertension were given antihypertensive treatment for 6 months and their blood pressures were tightly regulated and brought to the normotensive state. After 6 months, the estimations of the markers of oxidative stress were done again. RESULTS: The MDA levels were significantly increased in the stage I and stage II hypertension groups as compared to those of the control group (p<0.05). The antioxidant enzymes (SOD, Catalase and GPX) were significantly decreased (p<0.05) in the prehypertension and in the stage I and stage II hypertension groups as compared to those in the control group. There was a significant increase in the levels of the antioxidant enzymes after 6 months of a tight regulation and bringing of the blood pressure to the normotensive state by giving antihypertensive therapy. CONCLUSION: On comparison of the present study with other studies in which the use of antioxidants were found to be ineffective in the blood pressure reduction, it can be concluded that oxidative stress is an effect rather than a cause of essential hypertension.