Development of a diagnostic scar marker for Vibrio shilonii caused acute hepatopancreatic necrosis disease in whiteleg shrimp

Background: In a previous report, we showed that Vibrio shilonii was found on whiteleg shrimp (Litopenaeus vannamei) with acute hepatopancreatic necrosis disease in Thua Thien Hue province, Vietnam. This study was performed to develop a diagnostic molecular marker generated by random amplified polymorphic DNA (RAPD) for V. shilonii rapid detection.Methods: Pathogen Vibrio spp. were isolated from shrimps and fishes, and were identified by 16S rRNA sequencing. Genetic diversity of Vibrio strains was analysis by RAPD technique. Specific PCR product for V. shilonii was cloned and sequenced. SCAR marker was developed from specific PCR product.Result: Twenty random primers were evaluated for RAPD to identify DNA polymorphisms between Vibrio species. The random primer OPN-06 generated a 468-bp DNA fragment specific for V. shilonii. This was then converted into a sequence-characterized amplified region (SCAR) marker designated N6-441.Conclusion: Specific primers (Vshi-441F/Vshi-441R) amplified a unique DNA fragment in all V. shilonii isolates but not in the other Vibrio spp. This PCR assay showed significantly sensitive to the target DNA and reliably for the amplification the V. shilonii genome.Keywords: AHPND; RAPD; SCAR; Vibrio shilonii; Vietna

Analysis of mRNA Expression for Injury-Age Estimation

Background: Determination of wound age is pivotal in forensic medical sciences, criminal and civil cases for the construction of crime scene and answering the questions like, time of infliction, manner of wound infliction, how long the person survives after infliction of wound and characterizing antemortem or postmortem wounds. The findings differ considerably among individuals due to the biological variations. Previous developed method in the injury-age determination is clinical, microscopic, enzymatic reaction at the wound margins, histological, and immunohistochemistry with the pitfalls associated with it.Methods: This study is conducted on blunt injuries, particularly lacerated wound (type of wound inflicted by blunt weapon) to analyze the different expression pattern in injury-age up to 72hrs in total 21 individuals randomly grouped in different time intervals. To determine the time of injury, transcript abundance of mRNA of Fibronectin (FBN), IL1β, VEGFA, and GM-CSF was analyzed by real time polymerase chain reaction. 18S-rRNA was used as control marker.Results: Percent knockdown (%KD) was calculated to determine the expression of mRNA for the determination of injury-age. IL1β and GM-CSF showed the predictive behavior for wound age up to 36hrs, Fibronectin (FBN) showed predictive behavior up to 12hrs while VEGFA showed prediction beyond 72hrs.Conclusion: The detection of gradual decrease of mRNA of Fibronectin (FBN), IL1β, VEGFA, and GM-CSF may provide an estimation of wound-age.Keywords: mRNA Expression; Injury-Age Estimation; Antemortem wounds; Postmortem wound

Pomegranate juice attenuates acetaminophen induced hepatotoxicity in rat model of experiment

Background: Pomegranate fruit has high contents of various polyphenols and antioxidants due to which it possesses variety of therapeutic properties. In particular, pomegranate fruit peel and flowers have exhibited high antioxidant activity in different studies. Acetaminophen (APAP) overdose is the most common cause of drug induced liver toxicity including both accidental and intentional types. In our study, we investigated the protecting mechanism of pomegranate fruit juice (PJ) against toxicity caused by APAP in Wistar rats.Methods: Rats were fed with 0.2% (w/v) pomegranate fruit extract as prophylaxis to counter single dose of APAP (2 g/kg, p.o). After that variation in levels of glutathione, glutathione peroxidase (GPx), and glutathione reductase (GR) were marked.Results: A single dose of APAP elevated serum toxicity markers including lipid peroxidation. A simultaneous sharp depletion of glutathione and its metabolizing enzymes glutathione peroxidase (GPx), glutathione reductase (GR) was observed. Oral doses of PJ at (0.1% & 0.2% w/v) caused a significant (P<0.001) reduction in toxicity marker enzymes. A striking elevation in antioxidant armory was seen as in response to PJ.Conclusion: The results provide a clear picture of the defensive effect of PJ against APAP induced hepatic toxicity

Assessing the life quality in a Moroccan population of diabetics by the General Quality of Life Questionnaire SF36

Background: The quality of life assessment reflects the impact of chronic diseases on patients. The objective of our study is to assess the life quality of diabetic patients and to determine the factors that affect it.Methods: A cross-sectional study was conducted in 2019 in the Hygiene Service of the Sidi-Othmane district (Casablanca) in 526 diabetic patients. Quality of life was estimated using the SF-36 questionnaire to determine factors that affect mental or physical health. We used Pearson's correlation test for continuous variables and ANOVA for discontinuous variables.Results: Our patients have an average of life quality considering PCS and MCS respectively (38.47 ± 9.28; 41.15 ± 10.55). Association analyses showed on the one hand associations between the PCS and the gender (p = 0.0001), professional activity (p = 0.0001), marital status (p = 0.0001) and with chronic complications (p = 0.0001). On the other hand, between physical activity and PCS (p = 0.0001) and with MCS (p = 0.0001), between the level of instruction and the PCS and MCS dimensions respectively (p = 0.0001; p = 0.011) between acute complications and PCS and MCS dimensions respectively (p = 0.0001; p = 0.001). The correlation study showed inverse correlations between PCS and age (p = 0.0001), the diabetes duration (p = 0.0001), HbA1c (p = 0.037), and BMI (p = 0.008), and between MCS, and BMI (p = 0.0001).Conclusion: The presence of one of these factors (female, advanced age, high diabetes duration, complications, and unbalanced diabetes, low level of education, physical inactivity, and professional inactivity) leads to poor life quality.Keywords: Diabetes; Life quality; Physical health; Mental healt

MinION as a Teaching tool in a Graduate course in Pakistan

Genomics is a rapidly evolving field of study that is increasingly being utilized as a tool to detect ethnic and tribal-specific mutations that may be the key to rare and common diseases with higher prevalence in the population under study [1]. However, researchers and science educators in remote areas can often find it difficult to access the latest genetic technologies, probably due to its high costs and lack of suitable infrastructure. Recent technological innovations are resulting in portable, low-cost instruments that enable next-generation sequencing in remote environments, offering new opportunities to generate a more widespread network of trained geneticists [2]. We need to formalize educational efforts to teach students and young researchers with hands-on training to excel in molecular and bioinformatics knowledge. Here, we report our experience of using the MinION pocket size sequencer in a graduate course. The graduate course had some theoretical lectures that explained the basic principles of genomics followed be practical sessions. We hope that the training material developed during this course will provide the community with useful tools to help educate future generations of genome scientists in Pakistan.

Industrial Pesticides and a Methods Assessment for the Reduction of Associated Risks: A Review

Regarding the increasing growth of the population and importance of food security, Iranian Ministry of Agriculture has prioritized and encouraged greenhouse farming products. One of the developmental challenges of greenhouse farming is the current extensive use of chemical fertilizers. Importantly, as raw agricultural products are the main ingredients on the table of Iranian families  (Iranians generally tend to eat fresh products), the determination of pesticide residues in such products is of utmost importance. The penetration of resistant contaminants into freshwater resources can lead to detrimental effects on humans and the environment. Concerning the importance of environmental protection and the role of chemical pesticides, this study reviews the pesticides used in the agronomic sector and the associated risks of using chemicals to control pests for society, agriculture, freshwater resources, and the environment.Keywords: Environment; Water resources; Agriculture; Health; Biological metho

Phytotoxicity of Hg and its Detoxification through Microorganisms in Soil

Due to the advent of industrialization, pollution of terrestrial environment by heavy metals has emerged as a great issue. Therefore, it is an urgent need to realize the Hg-induced toxicity in plants and as well as in animals and the harmful effects by the consumption of contaminated nutrition. Mercury is considered as a hazardous contaminant that can be changed into various oxidation states easily and causes many deleterious effects in several physiological processes in both plants and animals. Microorganisms possess two extensively studied Hg-detoxification processes like Mer operon (merTPCFAD) and Met gene which encode the functional proteins for transportation (merT, merP and/or merC, merF), reduction (merA) and a secondary regulatory protein (merD) and sulfhydrylase enzymes (met gene) respectively to modify toxic Hg+2 to nontoxic elemental state (Hg0). Due to the ever increase in Hg-pollution and very little information about its phytotoxic effects and detoxification mechanisms, the authors expect, the present article will make possibility in the provision of  a comprehensive literature study about Hg-induced toxicity in plants and its detoxification processes to provoke for advance research in this field

Biosecurity and Biosafety concerns of Research and diagnostic Laboratory under International Guidelines

Currently, academic research labs and diagnostic laboratories are facing a serious issue of biosecurity and biosafety globally. The high rate of mutations and continuous new emerging infectious diseases with the risk of bioterrorism demands that each lab personnel share equal responsibility for biosecurity & biosafety at their work. The primary job of government authorities is to implement a well-organized detection system and limit the spread of hazardous biological agents by providing a biologically safe setting for lab scientists and for the common man in communities and institutes. The safety of lab personnel, lab environment, and pathogens depends on effective and safe laboratory working and pathogen handling which determines the reliable and accurate results of laboratory experiments. The aim of this article is to address the basic components of laboratory biosafety, laboratory biosecurity, and laboratory management. Further DURC (dual-use research of concern) deals with the commencement of lawful reasons to generate information, knowledge, technology, and products that are used for either harmful or beneficial purposes. The precautions are taken to ensure laboratory biosecurity and biosafety should be a part of the laboratory safety policy manual for guidance and implementation for a safe laboratory environment.Keywords: Antioxidants; tomato; lycopene; β-carotene; reactive oxygen species (ROS)

Biological movement of Xanthomonas oryzae pv. oryzae in Pakistan; A pioneer project of CEMB, Punjab, Pakistan

Background: In Asia, germ plasm exchange in different ecosystems, has been observed. This exchange causes movement of different pathogens. In present study, we worked on movement of Xanthomonas oryzae pv. oryzae (Xoo) causing bacterial blight of rice. In 1976, this disease was first recorded from Punjab (Pakistan) but the cultivars of that time showed resistance against bacterial blight. Then in 1984, the disease was seen on IRRI9. Tremendous increase of this pathogen has become prevalent due to increase in demand of Basmati 385 (cultivar), and now it is one of the most important rice diseases in Pakistan. Hypothesis of present study is “there is genetic diversity of Xoo in different rice growing areas of Punjab, Pakistan.”Methods: By fingerprinting; the movement pattern of Xanthomonas oryzae pv. oryzae were searched out in present study. The IS1112; a repetitive element of Xanthomonas oryzae pv. oryzae was used to fingerprint twenty-one bacterial strains.Results: We compared local strains with that of provided by IRRI Philippine (International Rice Research Institute, Philippine). The obtained clusters were correlated with regional differentiation.Conclusion: Present reported work is the pioneer study (1998-2000) which clued the occurrence of regional movement of pathogen via germ plasm exchange. This study may also provide help to forensics to watch the bioterrorism.Keywords: Bacterial blight; Xanthomonas oryzae; Germ plasm; Genetic diversity; SAARC

Association Of BCR-ABL Alternative Splice Variants with Disease Progression, Treatment Response and Survival in Chronic Myeloid Leukemia Patients Treated with Firstline imatinib Monotherapy

Background: Alternative RNA splicing has diverse biological effects in heath as well as disease. It also contributes to cancer onset and progression. Chronic Myeloid Leukemia (CML) results due to BCR-ABL fusion oncogene that is created due to chromosomal translocation t [9; 22] [q34; q11]). BCR-ABL is target of tyrosine kinase inhibitors (TKIs). BCR-ABL through alternative splicing can generate b2a2, b3a2 and some other rare splicing variants. BCR-ABL variants may vary in their response to TKI treatment and disease progression potential, which is a major factor contributing to dismal treatment outcome in CML. Objective: The objective of this study is to investigate correlation of BCR-ABL splice variants with TKI treatment outcome and survival in three phases of CML that has rarely been studied previously.Methods: BCR-ABL splice variants were studied using reverse transcriptase PCR (RT-PCR). in 70 CML patients from three phases of CML who were receiving imatinib (TKI) treatment.Results: Frequencies of different BCR/ABL splice variants like b3a2, b2a2 and b3a2+b2a2 were 49 (70%), 15 (21.4%) and 6 (8.6%), respectively. Splice variant b2a2 were more common (53.3%) in chronic phase CML (CP-CML) while b3a2 had higher frequency in advanced phases of CML (44.9%). CML patients with b2a2 transcript had better complete cytogenetic response and major molecular response to TKI treatment overall (100% vs. 24.5%) as well as in CP-CML (100% vs. 85.7%) and superior survival when compared to patients with b3a2 splice variant. All patients who died had male gender, less than 33 years age, b3a2 transcript, advanced phases of CML and imatinib resistance.Conclusions: Splice variant b3a2 was associated with CML progression, poorer survival and inferior treatment outcome as compared to b2a2. Further investigations on BCR-ABL splice variants and their roles in CML pathogenesis can provide deeper insights into CML biology and new targets for BCR-ABL positive leukemia treatment.          Keywords: CML; BCR-ABL splice variants; Progression; Survival; Treatment outcome