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Advancements in Life Sciences, volume 7, issue 1Cover; Volume 7, Issue 1
Published online: 25 November 2019
ISSN 2310-5380 

IN THIS ISSUE

 

 

Full Length Research Articles


Diurnal variation of brain derived neurotrophic factor and its importance
Muhammad Rafiq, Sajed Ali, pages 01-04
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 Background: 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. 

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Antioxidative properties of spices and their impact on postprandial blood glucose in humans
Abdul Rehman Khan, Nosheen Mushtaq, Humera Fiaz, Zia Ul Islam, pages 05-09

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 Background: 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. 

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Genotypic and computational sequence analysis of ALADIN gene causing Allgrove syndrome
Fezza Arshad, Nida Abdul Qadir, Nosheen Ishaq, Saqib Mehmood, Aiman Shehzad, Warda Fatima, pages 10-15

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 Background: Allgrove syndrome is autosomal recessive disorder, the gene involved in this syndrome is known as ALADIN located close to type 2 keratin gene cluster on chromosome 12q13 whose function is to control the nucleocytoplasmic trafficking and also affects the nuclear pore complexes. Mutation in this gene cause triple A syndrome. The aim of research was to analyze the mutational changes in ALADIN gene, formation of 3D Structure of normal and mutated protein and differentiation of normal and mutated protein.

Methods: Genotyping by using tetra arm PCR and Sequence analyses of coding region of ALADIN gene was done in two families having affected children with Allgrove’s syndrome.

Results: Point mutation in exon 1 and alteration in 3D structure of protein was observed by using VMD (Visual molecular dynamics) that shows truncation, absence of few amino acid and structural modification of proteins which alters in transportation ability.

Conclusion: It is concluded from the study that proper structure and function of NPC (nuclear pore complex) binding protein is necessary in normal body function and if any mutation is present in ALADIN gene it can cause symptoms of rare Allgrove’s syndrome. 

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Analysis of mRNA Expression for Injury-Age Estimation
Nusrat Saba, Saadia Noreen, Mubarak Ali Anjum, Saqib Ali, Muhammad Jawad, Allah Rakha, pages 16-19

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 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. 

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Arabidopsis Argininosuccinate Lyase and Argininosuccinate Synthase are important for resistance against Pseudomonas syringae
Shahbaz Anwar, Muhammad Amjad Ali, Amjad Abbas, Krzysztof Wieczorek, pages 20-26

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 Background: Arginine is one of the important amino acids and is involved in a variety of plant responses against biotic and abiotic stresses. Two important genes Argininosuccinate Synthase (AS) and Argininosuccinate Lyase (AL) are associated with the production of arginine from ornithine. Here we report molecular characterization of Arabidopsis thaliana artificial micro RNA (amiRNA) mutants for the genes AL and AS in response to infection with Pseudomonas syringae DC3000.

Methods: Quantitative real time PCR (qRT-PCR) and Promoter::GUS plant lines were used for expression analysis. Artificial micro RNAs (amiRNA) mediated gene silencing was used to generate mutant plant lines. Flood inoculation technique was used for infection test essays with Pseudomonas syringae.

Results: Expression analysis of A. thaliana plants harboring promoter AS::GUS construct showed strong promoter activity upon P. syringae infection. Quantitative real time PCR (qRT-PCR) analysis showed that AL and AS expression was strongly induced upon infection with P. syringae. Infection essays for P. syringae showed enhanced susceptibility to virulent (Pto) as well as avirulent (∆avrPto/∆avrPtoB) strains of P. syringae. However, mutant plants infiltered with infiltration medium containing 1 mM L-arginine regain their resistance in comparison with wild type (Col-0) plants.

Conclusion: Our findings suggest that genes related to arginine metabolism play a key role in plant defenses during P. syringae infection on A. thaliana. This study revealed that proper functioning of arginine related genes is required to deploy defense response against P. syringae. Decrease in the expression of these genes improves conditions for the growth of pathogen. 

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Genetic Analysis of SCN5A Gene in Patients of Long QT Syndrome
Uzma Zaheen, Memona Yasmin, Saadia Noreen, Saqib Ali, Allah Rakha, pages 27-31

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 Background: 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. 

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Mutational analysis of CYP1B1 gene in Pakistani pediatric patients affected with Primary Congenital Glaucoma
Muhammad Umer Khan, Raima Rehman, Haiba Kaul, Saqib Mahmood, Ali Ammar, pages 32-37

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 Background: Glaucoma is the significant cause of blindness all over the world. Primary congenital glaucoma (PCG) reduces the vision and ultimately causes the blindness by damaging the aqueous drainage system of the eye. The purpose of the current study was to determine the pathogenic mutations in the CYP1B1 gene responsible for PCG.

Methods: A total of thirty-five PCG patients were enrolled in this study. Blood samples were collected from the enrolled patients, and after DNA extraction and amplification, the coding regions of CYP1B1 were sequenced to determine the pathogenic mutations. In-silico analysis of the identified mutation was executed to study the effect of genetic variation on protein structure.

Results: One mutation, c.1169 G>A has been revealed in exon 3 of the CYP1B1 gene leading to p.R390H, present in 20% of the patients enrolled. Besides, two missense sequence variants c.1294G>C (2 patients), c.1358A>G (4 patients) and a synonymous variant c.1347T>C (18 patients) has also been observed.

Conclusion: Our study not only reaffirms the role of CYP1B1 mutations in PCG but also supports the use of genetic screening for molecular diagnosis and carrier identification, which will reduce the burden of disease on society. Furthermore, the in-silico analysis of the identified mutations provided an in-depth understanding of the PCG pathogenesis at the molecular level. 

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Genetic contribution of GJB2 gene to hearing impairment in Pakistan
Hamna Tariq, Kalsoom Zaigham, Samra Kousar, Aysha Azhar, pages 38-43

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 Background: Hearing impairment (HI) is defined as inability to hear and is an extremely heterogeneous genetic disorder. HI is divided into syndromic (if associated with clinical manifestation in addition to hearing impairment) and non-syndromic forms. So far one hundred and seventeen loci/genes have been mapped for non-syndromic HI and mutations in DFNB1 locus (GJB2 gene) are the most prevalent cause among them. This study was intended to find the relative contribution of the DFNB1 locus/ GJB2 gene for hearing loss in Pakistan and Azad Kashmir.

Methods: Twenty-one families were collected from different rural and urban regions of Pakistan and Azad Kashmir. The contribution of GJB2 gene was initially studied by linkage analysis using short tandem repeats (STR) microsatellite markers. Sanger sequencing was employed to identify the causative variants in coding region of the gene.

Results: Phenotype of four families were found linked with GJB2 gene and all affected individuals of these families segregating same mutation c.231G>A (p.Trp77*) which was confirmed after Sanger sequencing.

Conclusion: The genetic causes of hearing impairment were studied in twenty one families segregating autosomal recessive pattern of inheritance with different ethnicities. We further established the founder effect for the one recurrent mutation in GJB2 gene in Pakistani and Kashmiri hearing impaired families for the very first time. 

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In vitro anticoccidial activity of Trachyspermum ammi (Ajwain) extract on oocysts of Eimeria species of Chicken
Asghar Abbas, Rao Zahid Abbas, Muhammad Asif Raza, Muhammad Kasib Khan, Muhammad Kashif Saleemi, Zohaib Saeed, pages 44-47

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 Background: Due to development of synthetic anticoccidial drug resistance there is improvement of anticoccidial medicines due to which exploration of diverse therapeutic agents is attractive now a days. Amongst original agents’ miscellaneous botanicals have shown encouraging properties against coccidiosis. Present study is also a part of probing innovative therapeutic drugs against coccidiosis which can provide replacement solution to treat coccidiosis.

Methods: In current experiment in vitro anticoccidial effect of Trachyspermum ammi (seeds) extract was evaluated. For this purpose, an in vitro sporulation inhibition assay was used. Collected oocysts of four Eimeria species were exposed to six different concentrations (w/v) of T. ammi in 10% Dimethyl sulphoxide solution (DMSO), while Dimethyl sulphoxide and Potassium dichromate solution (K2Cr2O7) served as control groups.

Results: Results of study revealed that T. ammi extract showed in vitro anticoccidial effect by affecting on sporulation (%) and damaging (%) Eimeria oocysts in dose dependent manner. T. ammi extract also damaged the morphology of oocysts in terms of shape, size and number of sporocysts.

Conclusion: The results strongly support the botanicals applications of T. ammi extract and also demonstrate its potential for use in Poultry coccidiosis control strategies. 

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Review Article


Dry sockets – a systemic review
Bassam Mohammad Almutairi, pages 48-57

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 This is a systematic review, which shows a compendious review about the aetiology, prevention and treatment of Alveolar Osteitis. After tooth extraction the most common postoperative complication is dry socket. In oral surgery it is very essential to know about the pathophysiology aetiology, prevention and treatment of dry socket. The main purpose of this review article is to provide enough knowledge about the processes that are involved in the clinical management of dry socket. This article shows certain risk factors involved in the incidence of dry socket. surgical trauma, lack of operator experience, mandibular third molars,  oral contraceptives, menstrual cycles,  smoking, bacterial infections, excessive irrigation, age, bone fragments, local anaesthetics, we will discuss all of these factors which are involved in the incidence of alveolar osteitis in this article. Furthermore, some prevention methods have also been discovered which can be applied to prevent from the incidence of dry socket. Certain treatment methods have also been identified and the have played the vital role in treating the dry socket disease, this article also shows the relation between the Fibrinolysis and the occurrence of dry socket. Various treatment methods, which involve the use of antibiotics, chlorhexidine (as preventive measure also), steroids, anti-fibrinolytics, low-level laser therapy, have been identified which play a vital role in the treatment of dry socket disease. 

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