ARTICLE | doi:10.20944/preprints202101.0470.v1
Subject: Life Sciences, Biochemistry Keywords: Citrus fruits; patulin, dietary intake, variation in patulin; liquid chromatography
Online: 25 January 2021 (10:16:09 CET)
The research aims to discover the natural occurrence of patulin (PAT) in selected citrus fruits from the central cities of Punjab and Pakistan's Northern cities. Total 2970 samples of twelve citrus fruits; kinnow, orange, grapefruits, bitter orange, mausami, red blood, pineapple, sweet orange, rough lime, sweet lime, kagzi lime and lemon were examined using liquid chromatography fitted with UV detector. The limit of detection (LOD) and quantification limit was 0.04 and 0.12 µg/kg, respectively. The results have shown that 56% of samples of citrus fruits from Punjab's central cities, Pakistan, were discovered to be infected from PAT. The elevated amounts of PAT ranging from 0.04 to 1150 µg/kg were found in citrus fruit samples from Multan cities. Furthermore, 31.7% of samples of citrus fruits from Northern cities of Pakistan were discovered to be infected with PAT, and the elevated amounts were found ranging from 0.04-320 µg/kg from Swat city. About 22.1% of samples of citrus fruits have levels of PAT greater than the suggested limits established by the European Union (EU). PAT's dietary intake levels ranged from 0.10-1.11 µg/kg bw/day from the central cities of Punjab, Pakistan, and 0.13-1.93 µg/kg bw/day were documented from Northern cities of Pakistan.
ARTICLE | doi:10.20944/preprints202103.0695.v1
Subject: Chemistry, Analytical Chemistry Keywords: deoxynivalenol; corn; cultivar; grains; flour; bread; Pakistan
Online: 29 March 2021 (13:15:34 CEST)
The objectives of the current research were to determine the levels of deoxynivalenol (DON) in four different cultivars of corn and subsequently to investigate the fate of DON during pro-cessing steps involved for the production of cornbread. The samples (n = 30) of each cultivar which were found positive were selected for the study. The average level of DON was ranged from LOD to 650 µg/kg. The amount of DON in cornflour samples were ranged from LOD to 630 µg/kg and insignificantly lower than the levels found in corn grain samples (p ≥ 0.05). Further-more, the levels of DON in corn dough samples were insignificantly higher than the levels in cornflour samples (p ≥ 0.05), with levels ranged from LOD to 645 µg/kg. However, the amount of DON in cornbread samples was significantly different from the levels found in corn grains sam-ples (p ≤ 0.05), with levels ranged from LOD to 611.5 µg/kg. The percentage reduction of DON in grains to cornbread samples was 22.4%, 35.6%, 44.5%, and 42.6% in type 1, type 2, type 3, and type 4 cultivars, respectively. The highest dietary exposure and hazard quotient (HQ) of DON was 0.13 and 0.17 µg/kg bw/d, in male and female individuals resulted from the consumption of cornbread samples, respectively.
ARTICLE | doi:10.20944/preprints202103.0378.v1
Subject: Chemistry, Analytical Chemistry Keywords: DON; corn; corn products; exposure; risk assessment
Online: 15 March 2021 (12:04:05 CET)
The study focused on investigating the natural incidence of deoxynivalenol (DON) in corn and products from corn producing districts of Punjab, Pakistan. The analysis was carried out using HPLC with UV detector and immunoaffinity cleanup columns. The detection limit (LOD) and limit of quantification were 25 and 50 µg/kg, respectively. Total 1220 samples of corn and products were analyzed to detect the DON, and 539 (44.2%) samples were observed to be contaminated with DON (n ≥ LOD). Furthermore, 92 (7.5%) samples of corn & products have DON levels, elevated than the proposed limits of the EU. The data is significantly different from a normal distribution for DON in corn and products samples and from different locations (p < 0.05) for Shapiro-Wilk and Kolmogorov-Smirnov values. However, a significant difference in DON levels was found between corn and corn derived-products types (p ≤ 0.05). The lowest and highest exposure & hazard quotient (HQ) of 0.92 and 9.68 µg/kg bw/d were documented in cornflour samples.
ARTICLE | doi:10.20944/preprints202103.0652.v1
Subject: Chemistry, Analytical Chemistry Keywords: chitosan coating; edible coating; guava fruit; shelf-life; HPLC; flavonoids
Online: 26 March 2021 (10:12:24 CET)
Guava is a vital fruit worldwide, especially in Pakistan, and due to its nutritional value famous in each age group. Due to a very short shelf life, the marketing and export of this fruit faced severe constraints. Therefore, in the current study, edible coating of chitosan (0, 0.5, 1.0, 1.5, and 2.0%) was evaluated on postharvest shelf life when guava fruits were stored (room temperature and 4 °C temperatures) for 12 days. The chitosan treated coating fruits have shown reduced total sugars and malondialdehyde levels compared to untreated control samples. However, a significant difference (p ≤ 0.05) in total sugar and malondialdehyde levels exists between samples stored in m compared to refrigerated temperature (4 °C). The chitosan-coated samples have shown a greater amount of vitamin C, quercetin, rutin, and total phenolic contents than control samples. However, these nutritional parameters' levels were significantly different (p ≤ 0.05) in samples stored at room than samples stored at refrigerated temperature. However, the levels of crude fiber, potassium, and sodium were found statistically nonsignificant (p ≥0.05) in control versus chitosan treated coating treatments. The findings have documented that the coatings of 1.5 and 2.0% were most effective for extension in shelf life and maintaining the nutritional attributes of guava fruit.
ARTICLE | doi:10.20944/preprints202011.0486.v1
Subject: Physical Sciences, Acoustics Keywords: AFs; Edible seeds; Tocopherols levels; Dietary intake
Online: 18 November 2020 (23:07:40 CET)
A total of 779 samples of edible nuts (melon seeds, watermelon seeds, pumpkin seeds, and cantaloupe seeds) from southern cities of Punjab, Pakistan were collected from the summer and the winter seasons. The natural occurrence of aflatoxins (AFs) and vitamin E (tocopherols) levels were investigated using HPLC. The results have shown that 180 (43.4%) samples from the winter season and 122 (33.4%) samples from the summer season were found positive with AFs. The elevated average levels of total AFs (20.9±3.10 µg/kg) were observed in watermelon seeds without shell and the lowest average amount (15.9±3.60 µg/kg) were documented in melon seeds without shell samples from the winter season. The elevated average amount of total AFs 17.3±1.50 µg/kg were found in pumpkin seeds available as without shell. The results have documented a significant difference in total AFs levels in edible seeds available as shells versus without shells (α = 0.05 & 0.01). The highest dietary intake of 6.30 µg/kg/day was found in female individuals from pumpkin seeds (without shell) in the winter season and the value of 3.00 µg/kg/day were found in pumpkin seed without shell in summer season in female individuals. The highest amount of total tocopherol levels of 22.2 ± 7.70 ng/100g in pumpkin seeds samples from winter season and 14.5 ± 5.50 mg/100g were found in melon seeds samples from summer season. The variation of total tocopherol levels in edible seeds among the winter and summer seasons showed significant difference (p ≤ 0.0054), except watermelon seeds samples which shown non-significant difference (p ≥ 0.183).
ARTICLE | doi:10.20944/preprints202103.0681.v1
Subject: Chemistry, Analytical Chemistry Keywords: AFB1; AFs; vegetable seeds; vegetable oils; dietary intake
Online: 29 March 2021 (11:09:17 CEST)
A total of 744 samples of vegetable seeds and oil (soybean, sunflower, canola, olive, corn, and mustard) were collected for the presence of aflatoxin B1 (AFB1) and total aflatoxins (AFs). The results have shown that 92 (56.7%) samples of branded and 108 (57%) samples of non-branded edible seeds were observed to be contaminated with AFs. The maximum mean of AFB1 and total AFs in non-branded soybean seeds was 21.01 ± 4.70 and 36.37 ± 6.10 µg/kg, respectively. Furthermore, all samples of edible seeds have concentrations of AFB1 greater than the proposed limit of European Union (EU, 2 µg/kg) and 12 (7.40%) samples of branded seeds and 14 (7.40%) samples of non-branded seeds were found in the range ≥ 50 µg/kg. About 78 (43.3%) samples of branded edible oil and 103 (48.3%) sample of non-branded edible oil were observed to be positive with AFs, and the elevated average of AFB1 (14.29 ± 2.51 µg/kg) and total AFs (25.61 ± 7.50) µg/kg were found in non-branded soybean oil samples. Furthermore, 16 (8.88%) and 6 (3.33%) samples of branded vegetable oil have levels of total AFs in a range (21 - 50 µg/kg) and ≥ 50 µg/kg, respectively. The findings have indicated significant difference of AFs levels between branded and non-branded vegetable oil samples (t = 22.274 and p = 0.000) at α = 0.05 and significant difference of AFs levels in vegetable seeds and oil samples ( t = -17.75, p = 0.000) at α =0.05. The highest dietary intake was found in non-branded sunflower oil sample (0.90 µg/kg/day) in female individuals (16-22 age group), followed by the dietary intake of 0.69 µg/kg/day body weight in male individuals (16-22 age group).
ARTICLE | doi:10.20944/preprints202108.0498.v1
Subject: Medicine & Pharmacology, Oncology & Oncogenics Keywords: CML; Disease progression; common biomarker; drug target; ANRD36.
Online: 25 August 2021 (16:03:46 CEST)
Background: Chronic Myeloid Leukemia (CML) is initiated in bone marrow due to chromosomal translocation t(22;9) leading to fusion oncogene BCR-ABL. Targeting BCR-ABL by tyrosine kinase inhibitors (TKI) have changed fatal CML into an almost curable disease. Despite that, TKIs lose their effectiveness due to disease progression. Unfortunately, mechanism of CML progression is poorly understood and common biomarkers for CML progression are unavailable. This study was conducted to find out novel biomarkers of CML progression by employing whole exome sequencing (WES).Materials and Methods: WES of accelerated phase (AP-) and blast crisis (BC-) CML patients was carried out, with chronic phase CML (CP-CML) patients as control. After DNA library preparation and exome enrichment, clustering and sequencing was carried out using Illumina platforms. Statistical analysis was carried out using [SAS/STAT] software version 9.4 and R package employed to find mutations shared exclusively by all AP-/BC-CML. Confirmation of mutations was carried out using Sanger sequencing and protein structure modelling using I-Tasser followed by mutant generation and visualization using PyMOL. Results: Three novel genes (ANKRD36, ANKRD36B and PRSS3) were mutated exclusively in all AP-/BC-CML patients. Only ANKRD36 gene mutations (c.1183_1184 delGC and c.1187_1185 dupTT) were confirmed by Sanger sequencing. Protein modelling studies showed that mutations induce structural changes in ANKRD36 protein. Conclusions: Our studies show that ANKRD36 is a potential common biomarker and drug target of early CML progression. ANKRD36 is yet uncharacterized in human. It has the highest expression in bone marrow, specifically myeloid cells. We recommend carrying out further studies to explore the role of ANKRD36 in biology and progression of CML.