Advancements in Life Sciences (E-Journal, University of the Punjab)
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Trackable CEMB-Klean Cotton Transgenic Technology: Affordable Climate Neutral Agri-biotech Industrialization for Developing Countries
Get PDFBackground: Transgenic technology reflects the incorporation of novel useful traits in crop plants like cotton for economic benefits by overcoming the problems including insects’ pests and weeds in special. The present study is the success story of the continuous effort of CEMB team started back in the 1990s.Methods: This study includes characterization of a large number of Bacillus thuringiensis (Bt) strains taken from local soil and subjected to direct transformation of isolated BT genes into local cotton cultivars. Protocols for transformation into cotton plants were optimized and validated by the development of double gene codon optimized (Cry1Ac and Cry2A) transgenic cotton varieties.Results: The resulting GMOs in the form of CEMB-33, CA-12, CEMB-66 have been approved by Punjab Seed Council in 2013 and 2016 respectively. Double Bt and weedicide resistant cotton harboring CEMB-Modified and codon optimized cp4EPSPS (GTGene). These varieties can tolerate glyphosate spray @ 1900ml per acre without the appearance of necrotic spots/shedding and complete removal of all surrounding weeds in the cotton field is a significant advance to boost cotton production without spending much on insecticides and herbicides.Conclusion: In the current report, two unique sets of primers which amplify 1.1 Kb for CEMB-double Bt genes and 660 bp product for CEMB-Modified cp4EPSPS (GTGene) were tested. CEMB cotton variety CKC-01 is specially designed as low cost and easy to use by local farmer’s technology has the potential to revolutionize the cotton growing culture of the country.Keywords: Event detection; Bt Cotton; CEMB transgenic technology; GTGen
Genetic Analysis of SCN5A Gene in Patients of Long QT Syndrome
Get PDFBackground: Finding out the cause of death is the main concern in forensic casework, particularly in sudden unexplained death cases. Long QT syndrome is a form of life debilitating cardiac arrhythmia. Among LQT genes, LQT3 is suggested to be more lethal and patients are at high probability of sudden death as compared to LQT1 and LQT2 patient.Methods: Samples of long QT patients were collected and the coding regions of all exons of the SCN5A, voltage- gated Na+ channel gene were screened by sequencing for the potential mutations as the causative agent of long QT syndrome.Results: After data analysis, 3 genetic variations have been found, amongst them are 2 heterozygous mutations that were reported previously in other ethnicity G87A-A29A found in exon 2, G1673A-H558R in exon 12 and one novel mutation that has not been reported so far, G>A 1238-G412G resulting in a transition mutation and change in amino acid GGG- Glycine to GAG- glutamic acid at position 412 in exon 9. While in other exons, no significant mutation was found.Conclusion: As some of the exons showed mutation and the sample size was small, however, further functional analysis of the gene is needed with large number of samples for the confirmation of results.Keywords: Long QT Syndrome; SCN5A gene; Genetic variations; Sudden deat
Antibacterial activity of leaf extracts of seven grape cultivars against six strains of bacteria
Get PDFBackground: Grape (Vitis spp.) is one of the most widely grown fruit crops in the world. It contains large amounts of phenolic compounds which have antimicrobial properties. Many efforts have been done to discover new antimicrobial compounds from various kinds of natural sources to benefit mankind. The current study is aimed to evaluate the antimicrobial properties of the methanolic extract of different cultivars of grapes against six bacterial strains.Methods: In the present study, the antibacterial activity of different concentrations (25, 50 and 100mg/mL) of methanolic leaf extracts of seven cultivars of grape (Red Globe, Autumn Royal, Crimson, Thompson, Sundarkhani, Perlette and King’s Ruby) was tested against six strains of bacteria (Proteus sp., Streptococcus viridans, Escherichia coli, Pseudomonas sp., Staphylococcus aureus and Clostridium septicum) using agar well diffusion method.Results: The methanolic leaf extract of all the cultivars showed significant antibacterial activity, suppressing the growth of all the six bacterial strains tested. The inhibition zones against the different bacteria ranged from 12.6 mm to 29.6 mm. The highest zones of inhibition were produced by Proteus sp. (29.6 mm), Pseudomonas sp. (27 mm), Staphylococcus aureus (27 mm), E. coli (26 mm), Streptococcus viridans (23.3 mm) and Clostridium septicum (21.0mm) at 100 mg/mL concentrations of different cultivars.Conclusion: This study confirms that Vitis vinifera methanolic leaf extracts have inhibitory activity against all the tested microorganisms and is worthy of further investigation.Keywords: Vitis vinefera; Antibacterial activity; Proteus sp.; Streptococcus viridans; E. coli; Pseudomonas sp.; MRSA; Clostridium septicum; Agar well diffusio
Effect of yeast supplementation on production parameters, egg quality characteristics and crude protein digestibility in hens
Get PDFBackground: Yeast (Saccharomyces cerevisiae) has a buffering effect in the digestive tract, and proper feed additives in chicken diets can improve the birds’ digestive efficiency. The present study planned with aim to investigate the effects of different levels of yeast supplementation on the egg production, egg quality traits and crude protein digestibility in hens.Methods: A total of 84 hens (40 weeks of age) were fed the dietary treatments until 50 weeks of age. The hens were distributed into 4 treatments and 3 replicates with 7 hens per experimental unit. The yeast was incorporated into basal feed with various levels (0, 0.1, 0.15 and 0.2%). The production data was obtained during trial. For determining egg quality characteristics, 30 eggs from each treatment group were used at 14-day interval of the experiment from a 2-day collection of eggs during the week. The excreta from each replicate were collected after every 2 weeks for determining the crude protein digestibility.Results: Maximum egg production and egg mass were attained from the hens provided 0.15% and 0.20% yeast supplementation. The eggshell thickness decreased with supplementation of yeast. Higher yolk weight, albumen weight, Haugh unit and grading values were observed at 0.2% level than other groups. Crude protein digestibility also improved at high levels of yeast supplementation at all ages.Conclusion: This study envisaged that yeast supplementation between 0.15 and 0.20% could be sufficient during 40 to 50 weeks of age, to augment the overall layer performance. Keywords: Baker’s yeast; feed intake; egg production; egg qualit
Analysis of mRNA Expression for Injury-Age Estimation
Get PDFBackground: 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
Antioxidative properties of spices and their impact on postprandial blood glucose in humans
Get PDFBackground: Diabetes mellitus represents disrupted orderly processes of carbohydrate metabolism, in which body cells become unable to utilize glucose. Consequently, glucose molecules pile up in blood and exceed upper normal limit. Spices are vital source of bioactive molecules, which could be useful to treat different diseases including diabetes. Present study was conducted to determine the effect of turmeric, cloves, green cardamom and cinnamon intake on postprandial blood glucose (PBG) levels in normal healthy human subjects.Methods: A total of 10 participants including both genders were considered to assess blood glycemic response of said four culinary spices. Participants’ ages were 20-25 years. Incremental area under the curve (IAUC) method was employed for glycemic index (GI) determination. In addition to this, antioxidative properties were estimated by 2, 2-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid (ABTS+)) radical, Folin-Cioucalteau reagent and aluminum chloride.Results: GI values of turmeric, cloves, green cardamom, cinnamon and their combined blend were 83.06, 87.48, 82.27, 73.59 and 69.48, respectively. Antioxidative activity (AA) of spices was 2.63, 1.55, 2.55 2.8 and 3.33, respectively. Regarding antioxidant levels cinnamon contained the highest amount (32.78 mg/g) of total phenolic compounds (TPC) than turmeric (28.7 mg/g), cloves (29.6 mg/g) and green cardamom (15.04 mg/g). Similarly, total flavonoid contents (TFC) were found maximum (6.17 mg/g) in cinnamon relative to the other three spices i.e. 2.66, 4.6 and 1.6 mg/g, respectively. Furthermore, GI was inversely related to antioxidative properties i.e. AA (r=-0.88), TPC (r=-0.5625) TFC (r=-0.7716).Conclusion: The results obtained from this present study indicate that spices’ antioxidants interfere with gastrointestinal digestion, lowering starch conversion into blood glucose, effectively. An appropriate intake of spices may be wanted to keep blood glucose level within an optimum limit.Keywords: Antioxidants; Glycemic index; Humans; Spice
Diurnal variation of brain derived neurotrophic factor and its importance
Get PDFBackground: Previous studies have shown the importance of Brain Derived Neurotrophic Factor (BDNF) in different cognitive processes including learning and memory. Some previous studies have showed variations of BDNF protein during the day. But still there is no data that shows any circadian variation of BDNF in the brain of rodents. So, this study was aimed to determine any circadian variation of BDNF protein in brain structures involved in cognitive processes.Methods: Male Arvicanthis ansorgei diurnal rats were sacrificed at different zeitgeber times (ZT21, ZT17, ZT13, ZT9, ZT5 and ZT1). ZT12 and ZT0 defining lights off and on respectively. The brains were removed and brain homogenates were prepared from hippocampus and cortex tissues. The amount of BDNF protein was assessed using ELISA technique on the brain supernatants.Results: Both the structures i.e. cortex and hippocampus showed a circadian variation of BDNF protein. In cortex, two peaks were observed i.e. at ZT5 and ZT17. Post-hoc analysis showed a significant effect between ZT5 and ZT13 (P<0.05). Hippocampus, also showed two peaks i.e. at ZT9 and ZT21. Post-hoc analysis showed a significant effect between ZT1 and ZT21 (P<0.05).Conclusion: Our novel results showed that both brain structures of diurnal rodents follow a circadian rhythms of BDNF protein. This study provides a focus for designing experiments and techniques that are based logically how circadian rhythms of different proteins contribute in pathology and how we can treat.Keywords: Zeitgeber time; Cortex; Hippocampus; ELISA; Arvicanthis ansorgei; Circadian rhythms; BDN
Forensic Entomology: A Comprehensive Review
Get PDFDetermination of post mortem interval is done with various methods depending upon the condition of the dead body and the circumstances in which it is found. Ruling out foul play in unnatural deaths has been an enigma of a sort for forensic pathologists and scientists all over the world. The methods to determine the cause and manner of death keep on emerging with ongoing researches in the fields of forensic sciences. Many methods have been devised to determine cause, manner and specifically time since death. Forensic entomology is one such field that aids in determination of time since death, especially in putrefied corpses found in different habitats in mysterious unknown circumstances. The field keeps on emerging with the advent of new techniques on molecular level, including DNA analysis and identification based on entomological data that has diversified over the years. The standards and guidelines for entomological data collection and processing needed for implication on the crime scene must be revised now and again for assistance of crime scene investigators, scientists, entomologists and pathologists. Various factors must be considered while processing entomological data that affect post mortem intervals. The keen study of life cycles, groups of similar insects, valid and standard methods of insect collection, breeding and identification can give a lead on cause, manner, time, place and circumstances of unknown or unnatural deaths.
Production of Remedial Proteins through Genetically Modified Bacteria
Get PDFRecombinant DNA technology has created biological organisms with advanced genetic sequences and has been extensively used to express multiple genes for therapeutic purposes when expressed in a suitable host. Microbial systems such as prokaryotic bacteria has been successfully utilized as a heterologous systems showing high therapeutic potency for various human diseases. Bioengineered bacteria have been successfully utilized for producing therapeutic proteins, treating infectious diseases, and disease arise due to increasing resistance to antibiotics. Prominently E. colifound to be the most widely used expression system for recombinant therapeutic protein production i.e. hormones, enzymes and antibodies. Besides E. coli, non-pathogenic lactic acid bacteria has also been considered as an excellent candidate for live mucosal vaccine. Likewise, S. typhimuriumhas been deployed as attenuated type of vaccination as well as in treatment strategy of various cancers due to its ability of wide progression in tumors. The present article is a summarized view of the main achievements and current developments in the field of recombinant therapeutics using bacterial strains focusing on their usability in therapeutics and future potential.
Sequence Diversity of MAOA Gene within Wild and Docile Animal Species
Get PDFBackground: Molecular characterization of MAOA gene was performed to investigate aggressive behaviour within wild (lion, leopard, and wolf) as opposed to docile animal (sheep, goat) species living in different habitats, by undertaking sequence diversity analysis of this gene.Methods: The MAOA gene was partially amplified by PCR for wild and docile animal species. Amplified DNA was sequenced and then analyzed using BioEdit and Sequencher softwares, while multiple sequence alignment and phylogenetics analysis were conducted through MEGA software. Bioinformatics tool like Prosite scan, Motif Scan and Prot Param were used to study properties of mutant proteins of MAOA gene.Results: Different polymorphic sites were observed which included c.956, c.1063 in docile animals and c.2530 in wild animals. Phylogenetic analysis based on this candidate gene endorsed the existing taxonomy of subject animals, while bioinformatics tools explored the altered characteristics of mutant MAOA protein. Conclusion: The newly found polymorphic loci in wild and docile animals in this study could have a role in behavioral response and acclimatization within their peculiar habitats. This study also highlights the genetic diversity of MAOA gene, which will add knowledge to the existing animal genetic resource of Pakistan.