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New Issue Published; 9(1)

Advancements in Life Sciences, volume 9, issue 1Cover; Volume 8, Issue 4
Published: 25 May 2022
ISSN 2310-5380 

IN THIS ISSUE

 

Review Articles


Appraisal of various approaches to produce biohydrogen and biodiesel from microalgae biomass
Liaqat Zeb, Muhammad Shafiq, Munir Ahmad, Asaf Khan, Dawood Ahmad, Arshad Mehmood, pages 01-12
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 Background: Study of populations at genetic level marks high importance in terms of determination of population specific patterns. The study and analysis of population specific genetic patterns have wide ranging applications from medicine to forensic science. The study of mtDNA control regions gives the insight of maternal lineages of particular ethnic group and their evolutionary as well emigrational history. 

Methods: This research gives information of mitochondrial DNA haplotypes data of CR (control region) covering the area from 16024bp to 576bp of mitochondrial DNA of Wakhi population of Hunza valley from Pakistan. Samples of 40 unrelated Wakhi from upper Hunza were sequenced and their sequences encapsulating mtDNA control region was compared to rCRSs (revised Cambridge reference sequence) to see maternally inherited DNA variation at genetic level in this population.

Results: The results showed that all forty (40) haplotypes are unique. The haplotypes corresponded to 67.9% West Eurasian haplogroups followed by the Middle East and variety of Asian haplogroups exhibiting admixed maternal genetics of this population. Wakhi population comes with high genetic diversity (0.998) in turn lowest random match probability (0.026) and high power of discrimination (0.974).

Conclusion: This study gives interesting highlights important aspect of uniparental genetics of Wakhi population and is also a contribution to mtDNA control region data of Pakistani populations for applications in criminal investigations. 

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Full Length Research Articles


Genetic Characterization of Wakhi People from Hunza Valley of Pakistan by employing Mitochondrial DNA Control Region
Marriam Jaffer, Muhammad Saqib Shahzad, Zia Ur Rahman, Azam Ali, Saeeda Kalsoom, Javed Iqbal Bajwa, Muhammad Farooq Sabar, Alamgir Alvi, pages 13-17
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 Background: Study of populations at genetic level marks high importance in terms of determination of population specific patterns. The study and analysis of population specific genetic patterns have wide ranging applications from medicine to forensic science. The study of mtDNA control regions gives the insight of maternal lineages of particular ethnic group and their evolutionary as well emigrational history. 

Methods: This research gives information of mitochondrial DNA haplotypes data of CR (control region) covering the area from 16024bp to 576bp of mitochondrial DNA of Wakhi population of Hunza valley from Pakistan. Samples of 40 unrelated Wakhi from upper Hunza were sequenced and their sequences encapsulating mtDNA control region was compared to rCRSs (revised Cambridge reference sequence) to see maternally inherited DNA variation at genetic level in this population.

Results: The results showed that all forty (40) haplotypes are unique. The haplotypes corresponded to 67.9% West Eurasian haplogroups followed by the Middle East and variety of Asian haplogroups exhibiting admixed maternal genetics of this population. Wakhi population comes with high genetic diversity (0.998) in turn lowest random match probability (0.026) and high power of discrimination (0.974).

Conclusion: This study gives interesting highlights important aspect of uni-parental genetics of Wakhi population and is also a contribution to mtDNA control region data of Pakistani populations for applications in criminal investigations. 

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Mutational analysis of exons 5-9 of TP53 gene in breast cancer patients of Punjabi ethnicity
Haleema Sadia, Mukhtar Ullah, Asma Irshad, Kausar Malik, Waqas Ahmad, Rais Ahmed, Muhammad Umer Khan, Sana Ashiq, Ali Akbar, Atta-ur Rehman, Shahid Raza, Kashif Iqbal Sahibzada, pages 18-23

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 Background: This study was conducted to evaluate the determinants of anxiety among the people living in hassled environment for last few months due to outbreak of COVID-19.

Methods: 279 participants were interviewed through a questionnaire about COVID-19 anxiety on four likert scales. Data was analyzed by using statistical software IBM SPSS Statistics 21. A binary logistic regression was used to estimate relationship between anxiety and T.V news, social media news, social isolation, disturbance of sleep and decrease in daily income. Significance of these factors are tested at 5 percent level of significance.

Results: 90.33 percent of the respondents are feeling COVID-19 anxiety while 9.67 percent peoples are taking it mild. The values of test statistics for the variables listening of T.V news, social media news, social isolation, disturbance of sleep and decrease in daily income are highly significant at p < 0.05.

Conclusion: The existence of anxiety was found to be highly significance due to listening of T.V news, social media news, social isolation, disturbance of sleep and decrease in daily income.  

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Indigenous Curd as a Functional Food: A Source of Potential Pathogenic Bacterial Control
Shazia Yaqub, Mian Anjum Murtaza, Shinawar Waseem Ali, Sehrish Mushtaq, Sohaib Afzaal, Abdul Munim Farooq, Ghulam Mustafa, pages 24-31

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 Background: Dahi is a functional food that is largely consumed in our community.  The current study was designed to explore the efficacy of antibacterial agents that are present in the local indigenous curd of Lahore, Pakistan.

Method: Dahi was collected from various areas in Lahore. Bacterial cultures were isolated from this local curd and identified using a molecular technique that used 16S rRNA gene primers in PCR and sequencing. The antibacterial assay was carried out using the agar well diffusion method. Out of all the characterized bacteria isolate P26 displayed the best results for the control of pathogenic bacteria. Bacteriocin from isolate P26 was partially purified and its antibacterial assay was performed against sixteen indicator organisms and kanamycin was used as a standard for this experiment. Characterization of bacteriocin was performed at various temperatures, pH, and treating with enzymes.

Result: After the molecular characterization through 16S rRNA, Strain P26 was characterized as Bacillus vallismortis. A protease-sensitive antimicrobial substance was produced by Bacterial strain P26 (Bacillus vallismortis), isolated from local curd (Dahi). This isolate has a broad-spectrum inhibitory activity that can inhibit the growth of both food spoilage and pathogenic bacteria. According to results It was clear that protease-sensitive antimicrobial substance produced from strain P26 is more effective as compared to the antibiotic kanamycin. Maximum activity was shown against Staphylococcus species, a pathogen that is a common cause of gastroenteritis worldwide. This substance was active at 100 ºC for 30 minutes and pH (2-9). The activity was lost after treatment with proteases, so it is classified as bacteriocin.

Conclusion: Bacteriocin P26 is a more effective antibacterial agent than kanamycin, and it can be produced cheaply using local cardkey.  

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Antimicrobial Resistant Pattern of Isolates from Intensive Care Unit of Tertiary Care Hospital
Shehryar Ahmad, Ujala Rasheed, Iram Naz, Sajid Ali, Nasir Ali, Atif Aziz, pages 32-35

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 Background: Patients in intensive care units are more vulnerable and have a higher risk of Hospital acquire infection in comparison to non-critical patient’s wards. We determine the Microbial spectrum and their antimicrobial resistant pattern of isolates from intensive care units of tertiary care hospital in Peshawar, Pakistan.

Methods: A cross-sectional study was carried out for a period of 6 months and a total of 65 patients from the surgical intensive care unit and medical intensive care unit of tertiary care hospital were sampled for the possible pathogen. Non-probability convenient sampling techniques were used to enrolled patients and samples were transported to the pathology department for culture and anti-microbial resistant pattern. Data were analyzed on SPPS version 19 using descriptive and inferential statistical tools. Chi-square test were used for possible association and p value of < 0.05 were considered statistically significant.

Results: Both gram-positive and gram-negative isolates additional to fungi were prevalent in the intensive care unit with the most frequent isolates being Acinetobacter species followed by Enterobacteriaceae and  P. Aeruginosa respectively. Amikacin, Doxycycline, Linezolid, Tigecycline, and Vancomycin have a low resistant pattern for both Staph aureus and coagulase-negative staphylococci (CoNS) while Cephalosporin including cefepime (58%), and minocycline (35%) along with polymyxin B shows the lower resistance pattern for Acinetobacter and other gram-negative isolates. 

Conclusion: Tigecycline, vancomycin, cefepime, and polymyxin B were used as empiric therapy agents in severe conditions however a larger scale study is required for calculating optimized therapeutic regime for each agent. The physician and hospital guidelines should adhere to the monitoring of antimicrobials to limit the upsurge of resistance patterns among different pathogenic organisms.  

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Computational Prediction of Cassia angustifolia Compounds as a potential Drug Agents against Main Protease of SARS-nCov2
Rashid Saif, Kanza Ashfaq, Ghafran Ali, Ali Iftekhar, Saeeda Zia, Muhammad Zubair Yousaf, pages 36-40

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 Background: In November-December 2019, a plethora of pneumonia like cases were reported in Wuhan, China. After some time, the causative agent of this ailment was identified and named as a novel coronavirus 2. This novel virus spread over the world with no time and declared as pandemic by WHO. To develop antiviral drugs, different clinically used drugs were used as a trial but went in vain. In the current study, we choose an herb with already known therapeutic effects to check its antiviral properties against this virus too.

Methods: Cassia angustifolia is a well-known herb for pharmaceutical industries as its different compounds are already used in different medicines. Here we performed molecular docking of main compounds of Cassia angustifolia against the main protease of SARS-nCoV2 and were compared with different drugs that are already being used on commercial bases to obtain the lowest energy complex. Auto-Dock vina and its packages were used for molecular docking of SARS-nCov2.

Results: Molecular docking of Cassia angustifolia compounds represent very promising binding energies complexes, e.g., Sennoside B gives -9.05kcal/mol and Aloe-Emodin give -4 Kcal/mol of energy against the main protease of coronavirus. In contrast, a couple of commercially used antiviral drugs were also evaluated against the selected protein of coronavirus e.g., Hydroxychloroquine and Ribavirin complexes appeared with -5.2 Kcal/mol and -6.3 Kcal/mol of energy respectively.

Conclusion: Many compounds of Cassia angustifolia showed the promising energy complexes even better than the commercially used antiviral drugs e.g., Sennoside B which has the best energies against main protease of coronavirus. Further, in-vivo and in-vitro studies are needed to validate this hypothesis with advanced MD simulations and wet-lab experimentations. 

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Impact of supplemental exogenous lysolecithin on performance, fat digestibility, and lipid metabolites responses in broilers
Faizan Mahmood, Haq Nawaz, Sohail Hassan Khan, Muhammad Yousaf, Javid Iqbal, pages 41-48

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 Background: Coronavirus Disease-2019 (COVID–19) has appeared as a deadly pandemic affecting most of the countries across the world. The disease has caught humanity unprepared; therefore, there has been a lack of awareness about the causes, and transmission and mortality rates. It has also affected the overall socioeconomics from individual to worldwide levels. COVID-19 is an unprecedented and first-time challenge for Pakistan and a lockdown imposed by the Government of Pakistan has further devastated the economy of the country. This online survey was conducted to know the effects of the Covid-19 pandemic on the social life and economic status of the people of Pakistan. In this article, we mainly focused on teachers and students from different Universities of Pakistan for data generation because they can provide better information due to the use of modern technologies as a source of information.

Methods: An online questionnaire survey covering 11 parameters including age, gender, employment status, mortality rate, education, marital status and, information about the pathogen responsible for the disease was filled by 1260 individuals from Pakistan.

Results: The results obtained show that 56.6% of the responders were males,43.4% were females, 59.9% were unmarried, 37.7% were graduate students and, 48.7% unemployed. Most of the responders (55.6%) said that COVID-19 negatively affected the jobs of the people. We found (93%) of the responders with good knowledge about COVID-19.

Conclusion: We conclude from the results that most of the responders were males, educated and they knew the pathogenic effect of the COVID-19 virus on humans. These results also show that the disease has drastically affected the economy and social life of the people of Pakistan. 

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Study of Effect of Physiochemical Parameters on the Reproducibility (validation) of Biodegradation of Textile Dyes
Shagufta Ambreen Shaikh, Kausar Siddiqui, Nazia Masood, pages 49-53

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 Background: Environmental pollution due to different industrial waste has become a major problem in modern world because of their high toxic in nature. Textile wastes are carcinogenic and mutagenic contain varieties of toxic dyes and fixers. Azo dyes widely used in textile, paper and printing industries are not easily degradable and having carcinogenic nature. Due to health hazards of textile wastes, several chemical and biological methods are being used to make the discharged wastewater less toxic by degrading colors and other harmful chemicals. Present research study was aimed to evaluate the sensitivity of biodegradation process in form of Chemical oxygen demand (COD) and, to check the reproducibility of biodegradation results for validation purpose.

Method: Initially selective bacterial strains were isolated from different waste samples. COD of the three samples of dyes were performed by titration method to observe the pattern of reproducibility of the biodegradability (COD) results.

Results: Textile industry sludge wastewater samples contained different characteristics of dye degrading consortium of bacterial strains as compared to soil extracts samples whereas did not get any bacterial growth in domestic wastewater samples.  It was interesting to observe that the reproducibility of COD results was very hard to get due to sensitivity of the test parameter to the prevailing environmental conditions.

Conclusion: Present study is significant for understanding the sensitivity of reproducibility or validation of biodegradation study. On the basis of present findings of experiments, we can say that only the method of determining COD could be standardized which will ensure the credibility of the results and the changed environmental temperature could affect the controlled experimental results. Present finding would give benefit to the new scientists and students for understanding the normal behavior of biodegradation study. 

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A study on serological detection of Hepatitis A virus with associated risk factors in young kids
Aysha Iqbal Khan, Rana Haider Ali, Hafiz Muhammad Usman Siddiq, Suleman Irfan, Noor Fatima, Ramla Zafar, pages 54-59

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 Background: Hepatitis A virus (HAV) is a major concerning issue for human health that causes acute viral hepatitis. Hepatitis A virus is non enveloped RNA virus which is a member of the family Picornaviridae and genus Hepatovirus. HAV is more prevalent in developing countries like Pakistan with poor sanitation and economic status. The virus is present in young kids of age 1 to 15 years old associated with many risk factors.

Methods: A total of 100 blood samples (1-2 ml) were collected from the hospitals of Lahore. After serum collection, indirect ELISA was performed on a commercially available kit. The Optical density (OD) was taken from the ELISA reader. Positive and negative samples were also run along with the samples and samples OD was compared with the OD of positive and negative controls. Cut off value was calculated by multiplying negative control (NC) with 2.1. The absorbance value of specimen/cutoff <1: samples having a value less than cut-off value were considered negative and samples having a value greater than or equal to cut-off value were considered positive. The apparent prevalence of Hepatitis A virus was measured by dividing the number of children positive to the total number of children included in the study.

Results: Of the 100 samples tested, 37% were positive for IgM and 100% were positive for IgG. Statistical Analysis SPSS 21.0 version was applied to analyze the data and a correlation test was applied to see the association of risk factors with disease status.

Conclusion: This study was done to appraise the overall status of HAV prevalence in young kids in association with potential risk factors. 

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A study of PTF1 interaction with phosphorus stress inducing genes and its influence on root architecture of transgenic Arabidopsis
Aqsa Akhtar, Muhammad Irfan, Asma Maqbool, Kauser Abdulla Malik, pages 60-66

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 Background: Phosphorus; an essential macronutrient needed by the plant for its robust growth is inaccessible to the plant as required. Hence, a need arises to develop smart crops capable of utilizing maximum phosphorus from soil. PTF1(Inorganic Phosphorus Starvation Induced Transcription Factor 1) is overexpressed during phosphorus stress and regulates number of genes to combat this abiotic stress.

Methods: The current study is the first ever reported case of transforming Arabidopsis thaliana with plant expression binary vector pSB219 harboring PTF1 via floral dip method and analyzing phosphorus stress induced genes interaction, through yeast-one-hybrid. Yeast-one-hybrid analysis was performed on four selected genes namely LPR1 (Low Phosphate Response), PDR2 (Phosphate Deficiency Response 2), PHT1;2 (Phosphate Transporter) and RNS1 (Ribonuclease). The positive transformed lines were expression analyzed for PTF1 by real time PCR and further studied for their root morphology.

Results: The results clearly showed direct interaction of LPR1 with PTF1 while other genes, although being overexpressed, were indirectly regulated. Transformation efficiency of 1% was achieved and a maximum 2.5-fold increase in PTF1 expression was observed. Root morphological studies exhibited significantly enhanced root hair and lateral surface area when grown in phosphorus deficient MS medium.

Conclusion: The results of the current study may pave path for improved comprehension of gene interactions and root architecture modifications under phosphorus limiting conditions. 

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Investigations on molecular determinants of durable molecular response in chronic myeloid leukemia patients
Zafar Iqbal, pages 67-74

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 Background: Chronic myeloid leukemia (CML), a blood cancer, is caused by translocation between chromosomes 22 and 9 that gives rises to fusion oncogene BCR-ABL. In 20th century, CML was a deadly disease, but tyrosine kinase inhibitors (TKI) led to complete remission in over 80% CML patients. Nevertheless, TKIs are expensive, and discontinuation of treatment is required in patients with very stable treatment response. As no molecular markers of durable TKI response exist, this study was conducted to find out molecular determinants of durable response in CML patients treated with TKIs.

Methods: Peripheral blood and clinical data were collected from CML patients with durable treatment response, along with appropriate controls. DNA was extracted and whole exome sequencing (WES) carried out to screen novel genes mutated only in experimental groups and absent in control groups. Mutations were confirmed using Sanger sequencing.  Data was analyzed using SPSS version 23.

Results: Although WES detected 10 genes mutated exclusively in CML patients with durable treatment response, Sanger sequencing could confirm mutations only in RAI1 gene (GC deletion at nucleotides 837-838, a frameshift mutation).

Conclusions: Our study shows that mutations in a novel gene (RAI1) are associated with durable response in CML patients.  RAI1 gene is active throughout the body and controls functions of many genes involved in daily rhythms. Our studies provide first important insights into molecular factors associated with long-term treatment response in CML that can serve as novel biomarker to identify patients eligible for TKI cessation in many ongoing CML STOP-TKI trials. 

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Thyroid Dysfunction and Vitamin D Deficiency among Females of Punjab, Pakistan; A Cross Sectional Analysis
Jamil Ahmad, Altaf Hussain, Lubna Humayun, Muhammad Ahtisham Azhar, Fariha Zafarullah, Irfan Ullah, Jamal Shah, pages 75-79

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 Background: Thyroid gland is an important gland which plays a vital role in the stimulation of normal growth and central nervous system (CNS), metabolism regulation, elevated vitamin requirements, metabolism of phosphorus and calcium, promote sexual metabolism, increases mitochondrial metabolism, stimulates the adrenergic activity with myocardial contractility and increase heart rate. In this era, one of the important health issues is vitamin D and calcium deficiency. A large number of populations in all over world are vitamin D or calcium deficient or insufficient. The objective of this study was to evaluate the prevalence of thyroid dysfunction and vitamin D deficiency in females.

Methods: A cross sectional study was designed to check the prevalence of thyroid disease and its correlation with vitamin D levels in females. Venous blood was drawn from the female’s patients (11—80 years of age) using gel disposable vials (3.5 ml) in aseptic condition. Samples were centrifuged at four thousand revolutions per minute for five minutes and serums were separated. After the separation of serum, the samples were transferred to the laboratory for the automated estimation of Thyroid Stimulating Hormone (TSH), Triiodothyronine (T3), Thyroxine and Vitamin D.

Results: It was observed that out of 79 females who had gone through thyroid profile testing, 70% females had normal thyroid profile. However, hypothyroidism was found in 23% females and 7% females had the condition of hyperthyroidism. Out of 18 (23%) reported cases of hypothyroidism, 8 cases were of mild (subclinical) hypothyroidism, 3 cases indicated non-thyroidal illness; rare pituitary hypothyroidism. Out of 79 female participants, only 20 (25.31%) females had normal serum vitamin D levels. Overall, 59 (74.68%) females had vitamin D deficiency. When the vitamin D deficiency was correlated with thyroid dysfunction, it was observed that vitamin D levels were non-significantly (p = 0.35) associated with hypothyroidism.

Conclusion: Hypothyroidism was found prevalent in the tested female population, as 23% of the tested population had hypothyroidism while 9% of them have hyperthyroidism. Moreover, majority of the population had vitamin D deficiency. 

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Investigation on Possibility of Rodents to Transmit Human Ascaris lumbricoides eggs from Contaminated Soil
Mehru Nisha, Athira Mohd Ghozi, Aina Syahirah Ali Baderudin, Pang Jyh Chyang, pages 80-84

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 Background: Ascaris lumbricoides is commonly found in soils among communities living with poor sanitation facilities. Commonly human helminths do not survive in animals’ guts and vice versa. However, the high burden of Ascariasis among indigenous communities could be transmitted by rodents as a vector. Hence, this study was carried out to address this hypothesis. 

Methods: The eggs of Ascaris lumbricoides were isolated from soil samples from an indigenous village in Malaysia, the eggs were isolated using the floatation technique and cultured in vitro using 0.1% of sulphuric acid dish and embryonation of the parasite eggs was observed daily under a light microscope till day 28. For the in vivo method, the embryonated eggs around 400 epg and 800 epg were feed to the rats for four weeks. Finally, stool samples were collected for Ascaris egg shedding.

Results: The loamy eggs collected near toiled areas using the floatation technique were found to contain Ascaris lumbricoides eggs. The cultured eggs in sulphuric acid developed from the first-cell stage until the fourth stage of development. Discharge of Ascaris eggs were found in stool samples among rats fed with 200epg of Ascaris eggs.

Conclusion: It was found that 0.1% sulphuric acid was ideal for developing Ascaris lumbricoides eggs for cultivating in vitro condition, and there was the possibility of rats carrying the Ascaris eggs throughout the village. In future, these findings can be used to propagate the eggs for testing anthelminthic drugs inhibiting the developmental stages. 

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Sibling and non-sibling fingerprints comparison of Pakhtun population of Swat district, KP, Pakistan
Subhanuddin, Noor Ullah Khan, Murad Ali Rahat, Aftab Ahmad, Fazal Akbar, Naseer Ullah, Muzafar Shah, Akhtar Rasool, Muhammad Israr, pages 85-91
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 Background: Fingerprint and other ridges are considered to be the best forensic science tool for identification of humans, alive or dead, and even for decomposed bodies. These fingerprint ridges exhibit various static features throughout life which reflect the person biology. This branch gained immense importance since the past few decades in congenital abnormality. This study was to carry out fingerprints analysis of sibling and non-sibling for differentiation and gender identification.

Methods: A total of 80 pairs of fingerprints (1600 prints) were collected from persons aged 15 to 30 years using rolling method. Out of which 20 pairs were brother-brother, 20 were sister-sister, 20 were brother-sister and 20 Pairs were random. Each fingerprint was analyzed for the gender identification on the basis of minutiae, ridge density and types. All the fingerprints were analysed using ACE-V method. After comparison SPSS software was used for further analysis.

Results: Our result showed that the types of fingerprints identified was whorl (50%) followed by loop (45.25%), arch (4.5%) and 0.25% of the accidental type. The dominant type was whorl while accidental was the least common type of fingerprints. Statistical analysis showed that between the groups, brother-brother and sister-sister was significant while rest of the groups was not significant. Moreover, greater ridge density was observed in female as compared to male.

Conclusion: It is concluded that the sibling fingerprints had greater similarity as compared to non-sibling, however both male and female fingerprints were significantly different in term of ridges density. This study may be useful in crime scene investigation.  

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Evaluation of mitigating effects of salicylic acid against various levels of salinity in onion (Allium cepa)
Rashda Naheed, Noreen Akhtar, Muhammad Shahzad Afzal, Fozia Farhat, Muhammad Umer Farooq, pages 92-97
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 Background: Onion (Allium cepa) is a vital vegetable as well as medicinal crop grown all over the world. A considerable area in Pakistan is under salinity stress which markedly reduce the average yield of many crops. Salicylic acid is an important growth regulator and has mitigating effects against saline adversities.

Methods: A study was conducted in complete randomized design (CRD) using two onion varieties, Phulkaara and Desi Red, under Faisalabad, Punjab conditions. The level of salinity was kept 50 mM NaCl and 100 mM NaCl while 400 ppm of salicylic acid was applied weekly. The data was analyzed through Statix 8.1 to compute analysis of variance (ANOVA).

Results: All the growth and physiological parameter of onion seedlings were decreased significantly with the increasing level of salinity while increased with the application of salicylic acid except total soluble protein. Salicylic acid (@400ppm) mitigated the adverse effects of salinity by enhancing photosynthetic activities. The total soluble protein was increased in leaves with increasing salinity stress whereas the application of SA has non-significant effects. The performance of both varieties remained significantly same for the parameters dry shoot weight, plant height, chlorophyll a, b and total chlorophyll contents. The variety Desi Red performed better for shoot length while Phulkaara was better in fresh shoot/root weight, length of root, dry root weight and total soluble protein.

Conclusion: The application of foliar spray of salicylic acid is potential growth regulator to increase the growth parameters of onion under saline stress conditions.  

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Systems genomics of nucleoporins provides prognostic insights into breast cancer
Rashid Mehmood, Kazuya Jibiki, Zakeya J. Alsafwani, Muhammad Naseem, Noriko Yasuhara, pages 98-110
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 Background: Nucleoporins (Nups) constitute a large group of proteins that are structurally arranged at the nuclear envelope and facilitate the bidirectional movement of molecules across the nuclear membrane. In addition to regulating the shuttling of ribonucleoprotein complexes, RNAs and proteins, various Nups interact with chromatin either directly or indirectly, thus regulating gene expression. Any mutations or expression anomalies of Nups may lead to abnormal localization of critical molecules, or dysregulated expression of genes that they interact with. A comprehensive genomic study encompassing all Nup genes in relation to breast cancer is lacking.

Methods: We used genomic and transcriptomic datasets from Pan-Cancer TCGA (The Cancer Genome Atlas), Genotype-Tissue Expression (GTEx) and microarray platforms and conducted in silico analysis of all the genes encoding nucleoporins that are associated with the Nuclear Pore Complexes (NPCs).  For mutation detection, we used cBioportal; for expression analysis, we used Xena and for patient survival plots, KMPlot was utilized.

Results: The genetic and molecular profile of nucleoporin genes identified multiple mutations and detected aberrant expression in breast cancer. Interestingly, NUP133AHCTF1TPRNup121L showed simultaneous gene amplification in nearly 10% of breast cancer patients. In addition, deregulated expression of some of the nucleoporins, namely, NUP62, NUP 93, NUP98, NUP155, POM121L12, RAE1, SEC13, TPR were correlated with patient prognosis.

Conclusion: The current study is the first one that unravels a comprehensive molecular and genetic profile of nucleoporins genes in breast cancer and underscores the critical roles of various nucleoporins in cancer progression. The identified molecules may advance our understanding of the etiology of the disease and serve as possible targets for novel therapeutic strategies in cancer. 

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Molecular genetic analysis of the m.A3243G mutation of the tRNALeu (UUR) gene in a population of Moroccan deaf diabetics
Taha Rhouda, Fatima Zahra Kehailou, Fatima Zahra EL Mskini, Ali Labriji, Mohammed Jabari, Houriya Mestaghanmi, pages 315-319
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 Background: The mitochondrial DNA (mtDNA) m.A3243G mutation of the tRNALeu (UUR) gene presents clinically heterogeneous phenotypes and is often responsible for diabetes, with or without deafness syndrome. The aim of this study was to search for this pathogenic mutation in 3 diabetics with bilateral deafness and a family history of diabetes.

Methods: mtDNA was extracted from the patients' whole blood. After PCR amplification, the DNA was sequenced and analyzed.

Results: The sequencing results showed the absence of the most common mtDNA mutation m.A3243G in diabetic families with individuals who suffer from hearing loss.

Conclusions: The m.A3243G mutation in the mitochondrial tRNALeu (UUR) Leu gene was not found to be a common cause of type 2 diabetes and deafness. These results suggest that there may be genetic causes for this phenotype.  

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Antibacterial Evaluation of The Roots of Moroccan Aristolochia longa Against Referenced Gram-positive and Gram-negative Bacteria
Latifa Doudach, Samiah H. Al-Mijalli, Emad M. Abdallah, Hanae N. Mrabti, Fatiha Chibani, Moulay El Abbes Faouzi, pages 116-121
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 Background: Interest in medicinal plants has recently risen as a consequence of its therapeutic capabilities, which require further investigation. Aristolochia longa (A. longa) roots are commonly employed in traditional Moroccan medicine as an anticancer agent and against some cutaneous infections, but little is known about their antibacterial capabilities.

Methods: The disc-diffusion and minimal inhibitory concentration (MIC) methods were carried out to determine the antibacterial activity of aqueous and methanol extracts of A. longa roots against several reference bacterial strains.

Results: Both aqueous and methanolic extracts had antibacterial activity to varying degrees with the disc-diffusion assay, however the methanolic extract shown significant activity at a higher dosage (200 mg/ml). Escherichia coli (22.0 mm) was the most sensitive bacterium tested, followed by Staphylococcus aureus (16.0 mm) and Bacillus subtilis (12.0 mm). The bacteria with the lowest susceptibility were Klebsiella pneumoniaPseudomonas aeruginosa, and Micrococcus luteus, which all had a 10.0 mm zone of inhibition. The MIC values corroborated these observations, with E. coli MICs of 6.25±1.5 mg/ml, S. aureus MICs of 25±1.02 mg/ml, M. luteus MICs of 12.5±1.25 mg/ml, K. pneumonia MICs of 50.0±0.75 mg/ml, and B. subtilis MICs of 100±0.6 mg/ml, respectively. The findings of the methanolic extract were statistically significant and equivalent to those of the antibiotics studied.

Conclusion: Roots of A. longa have significant broad-spectrum antibacterial agents, notably in the methanolic extract, which validated their applications in Moroccan folkloric medicine to treat soft-tissue and skin infections.  

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A comparison of human blood preservation methods in DNA extraction protocol
Jawdat N. Gaaib, Zainab W. Juda, Raghda H. Aedan, pages 122-125
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 Background: Blood preservation method plays an important role in DNA extraction. The current study was conducted at the laboratories of the applied medical sciences college/ University of Karbala during the period from March to May 2021.

Materials: Ten Fresh venous blood samples were collected from healthy males (20-45 years old) by venepuncture and stored at 2–8oC until used. Each particular sample was subjected to five blood collection options as follows: the blood was freshly aliquoted without any treatments, the blood was collected in EDTA (ethylene diamine tetra acetic acid) tubes, the blood was collected with the addition of phosphate buffered saline (PBS) in a ratio of 1:2, the blood was added to PBS 1:2 and Triton X 100, and the blood was centrifuged and then PBS 1:2 and Triton X 100 were added. The quantity and integrity of the extracted DNA were evaluated by agarose gel electrophoresis. The quantity and purity of the DNA were measured using a NanoDrop Spectrophotometer.

Results: The current study indicated statistically significant difference (p ≤0.05) among the treated groups when the concentration of the extracted DNA was taken into consideration. There was a statistically significant difference among the five groups when their absorbance ratio was measured at 260 nm/ 230 nm (p ≤0.05). However, no statistically significant difference (p ≤0.05) was obtained among the groups when the absorbance ratio of 260 nm/280 nm was considered.

Conclusion: The pretreatment of blood samples with buffers (PBS containing 5% Triton X100) prior to DNA extraction can lead to an altered DNA yield and purity. 

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