Volume 11, Issue 2

Cover; Volume 11, Issue 2Advancements in Life Sciences, volume 11, issue 2 – May 2024
ISSN 2310-5380

IN THIS ISSUE

This 1st issue of the 11th volume has featured 39 multidisciplinary articles including 1 short communication, 3 review articles and 35 research papers of authors from 15 countries covering aspects from different stems of life science. Enjoy reading!

 

 

Short Communication


Elucidating Severity of Waves of COVID-19 in Pakistan
Rabia Zia, Zilwa Mumtaz, Zubia Rashid, Ashaq Ali, Muhammad Zubair Yousaf, pages 276-280

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 Background: COVID-19 has radically changed the globe. This virus has a powerful impact on almost every country in the world. The most significant hurdle arises for the overall capacity of a disease control when the number of cases peaks during each wave of the pandemic. There are various waves that have occurred in the preceding period, each of them is characterized by a distinct variant of the virus. For a developing country like Pakistan COVID-19 ruins every sector of the country. There is a proper need for care and protection from this virus as guided by the World Health Organization (WHO). The spread rate is different in every country based on various factors. Two or more factors are required to trigger a high infection rate.

Methods: The data sourced from the National Command and Operations Center (NCOC) was focused on analyzing the data from the four waves of COVID-19 in Pakistan encompassing reported cases, death and testing capacity. Utilizing the SPSS software, a comprehensive examination of these factors was performed.

Results: The data of four waves has been analyzed including reported cases, no. of death, and testing capacity of the country. The data analysis reveals there is a strong correlation between these factors. By the interpretation of the data, it reveals upcoming waves are more dangerous than earlier due to new mutations.

Conclusion: The virus is evolving to become more contagious and infectious in each consecutive wave. Vaccination should be considered compulsory for everyone to win the battle against the virus. Data analysis revealed a correlation between the number of cases and passage of time, showing that the new waves will emerge earlier and have a significant impact on the country. 

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 Review Articles


Immunomodulatory Role of Cytokines in Periodontal Disease

Batool Hassan Al-Ghurabi, Maha Adel Mahmood, Zainab A. Aldhaher, pages 281-285
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 Periodontal disease is an inflammatory condition that affects the periodontium and results in the destruction of periodontal tissue. Cytokines are small, non-structural proteins with inferior molecular weights which have an intricate organizational effect on inflammation and immunity. Immune reaction against products of bacteria and the resulting generation of inflammatory cytokines are significant contributors to periodontal tissue injury. In this disease inflammatory cytokines are generated by immune cells to attack the periodontal bacteria. A persistent or strong immune response may cause inflammation and the creation of cytokines that are essential in the development of periodontitis, even though the immune response can defend an organism from a variety of diseases. Clarifying the immunomodulatory role of cytokines in periodontitis was the main goal of this review. Using suitable keywords, relevant papers would be searched in the scientific databases Scopus, Google Scholar, PubMed, and Web of Science. In conclusion cytokines control and modulate the immune response in periodontitis. Covered studies were posted between 2000 and 2023 and detect great variance in patients’ selection, clinical assessments, and cytokines measures. A significant association between cytokines and periodontitis developments was reported in some studies. Cytokines are control and modulate the immune response in periodontitis. 

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Role of Reactive Oxygen Species in the Pathogenesis of Bronchial Asthma and Obstructive Pulmonary Diseases: Systematic Review
Imetkul Ismailov, Romanbek Kalmatov, Baktyiar Abdurakhmanov, Ali Munir Mirza, Jitendra Kumar Chaurasia, pages 286-295
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 The article discusses the role of reactive oxygen species (ROS) in the pathogenesis of bronchial asthma and obstructive pulmonary diseases. The review follows a structured approach beginning with a comprehensive search of electronic databases, then findings from the included studies were summarized and used to draw comprehensive conclusions. The authors state that increased activity of oxidative processes and insufficient activity of the antioxidant system lead to the accumulation of ROS in the respiratory tract. This leads to direct damage to epithelial cells, activation of inflammatory cells, and stimulation of inflammatory mediators. ROS also activate transcription factors that increase inflammation and airway obstruction. Thus, maintaining the balance of the oxidant-antioxidant system is an important strategy for the treatment of these diseases. 

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Current advances on single or multi-omics analysis of esophageal cancer
Kaidirina Kasimu, Wenwen Cui, Yihan Wang, Xin Li, Hongbo Wang, Xiaotong Yu, Fu Ren, pages 296-304

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 Esophageal cancer is associated with high mortality rates and is one of the cancers with the worst prognosis. Its incidence has significant regional specificity, particularly in China where it is much higher than in other countries. Moreover, effective diagnostic markers, therapeutic targets, and molecular subtyping biomarkers are currently lacking for esophageal cancer. Nevertheless, large-scale omics studies have identified dozens of robust genetic risk loci and prognosis-related loci, drawn genomic, epigenomic, and transcriptomic maps of esophageal cancer at multiple molecular levels, and described significant differences between esophageal squamous cell carcinoma and adenocarcinoma. These studies are of great significance for exploring the occurrence and development mechanism of esophageal cancer, guiding clinical treatment, and improving patient prognosis. This review, from the perspective of multi-omics, discusses the analytical strategies employed in these studies and summarizes their core findings. It emphasizes that the integration and analysis of multi-omics data is a key focus and development trend in the precise medical research of esophageal cancer, and has broad research and application prospects.  

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Full Length Research Articles


Investigation of Anthelmintic Property of Cow Urine Distillate on Ascaris and Trichuris eggs Isolated from Soil
Mehru Nisha, Ruziana Yusri, Pang Jyh Chyang, pages 305-308

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 Background: Helminth infection occurs due to contact with contaminated soils and poor hygiene practices. Even though there is immense progression in human medicine, infections that occur due to bacteria, fungi, viruses, and parasites persist as a crucial threat to public health. Over the years, many studies indicated cow urine has antimicrobial activity. Hence, this study looks into anthelminthic property of cow urine in Ascaris spp., and Trichuris spp., eggs. 

Methods: The soil sample was collected at the riverside, near the toilet area and at the pond area at an aborigine village in Malaysia. Next, floatation technique was used to isolate the helminth eggs and it was identified using a light microscope. The cow urine was prepared as raw cow urine (RCU), evaporated raw cow urine diluted with distilled water (ERCUD), evaporation raw cow urine diluted with saline (ERCUS) and evaporated filtered cow urine diluted with saline (EFCUS).

Results: From the soil samples, mainly two types of helminths were found which were Ascaris lumbricoides and Trichuris trichiura. Evaporated cow urine diluted with saline showed the most prominent result in the destruction of helminth eggs.

Conclusion: Evaporated cow urine possesses biochemical agents that can be used in the future as anthelmintic agents whereby further studies can be made to explore more possibilities to use cow urine as a potential anthelmintic agent. 

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Severity Assessment and IL-5 Evaluation after Immunotherapy in Atopic Dermatitis Mice
Sylvia Anggraeni, Dinar Chieko Triesayuningtyas, Hamidah Luthfidyaningrum, Made Putri Hendaria, Yuri Widia, Anang Endaryanto, Cita Rosita Sigit Prakoeswa, pages 309-314

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 Background: The effectiveness of immunotherapy in atopic dermatitis (AD) has been debated for many years. Immunotherapy suppresses Th2 cytokines, such as IL-5, which play a crucial role in the pathogenesis of AD. This study assessed the effect of immunotherapy on severity and IL-5 expression in AD mice.

Methods: Male BALB/c mice were separated into three groups. The mice were sensitized with Dermatophagoides pteronyssinus allergen for seven days, except the mice in control group. The house dust mite (HDM) immunotherapy was injected subcutaneously every 3 days for 1.5 months with increasing doses (0.1, 1, 10, 100 μg in 100 μL PBS) every 4 injections. The mice in AD model and control group received placebo injections. Following immunotherapy, the mice were exposed to HDM allergen patch two times with 2 weeks interval in between. The mice were evaluated for severity score as the clinical marker and IL-5 expression with semiquantitative method as the histological marker.

Results: The evaluation of severity score from two independent researchers showed a substantial agreement (Cohen’s kappa 0.613, p <0.001). The severity score of the immunotherapy group was significantly lower than the AD model group, while both immunotherapy and AD model group had significantly higher score than control group. IL-5 expression in the immunotherapy group was lower than the AD model group and slightly higher than control group. The mean difference between groups was not significant.

Conclusion: The severity of skin lesion and IL-5 expression in AD mice receiving immunotherapy were lower than AD model group. 

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Protective immune response of recombinant Fiber-2 protein as subunit vaccine against Fowl-adenovirus-4 infection in Pakistan
Muhammad Ashir Zia, Muhammad Salahuddin Shah, Mudasser Habib, Faiza Ather. Ali, pages 315-321

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 Background: Hydropericardium Syndrome (HPS) has emerged as a major viral disease of poultry industry causing huge economic losses in the recent past years. Its etiological agent, Fowl adenovirus-4 (FAdV-4) has been reported in the various countries including Pakistan. Various strategies including inactivated vaccines, are being devised to control the disease since its appearance. Subunit vaccines based on viral structural proteins have demonstrated more promising protection against FAdV-4 infection than commercial inactivated vaccines. Among different viral structural proteins, Fiber protein (mainly Fiber-2) has been as a suitable candidate for developing the recombinant subunit vaccine against HPS.

Methods: Considering the importance of the Fiber-2 gene, the pET28a expression vector was utilized to clone its open reading frame, which was subsequently expressed as an oligo-histidine tagged fusion protein in BL21 cells of Escherichia coli via the IPTG induction method. The expressed recombinant fiber-2 protein was purified using nickel (Ni2+) affinity chromatography and used as a subunit vaccine in broiler birds following challenge with pathogenic FAdV-4 isolate. The immunological response was evaluated using ELISA.

Results: The gene for the Fiber-2 protein was effectively cloned and expressed as a soluble 60 kDa protein, as detected by SDS-PAGE and western blot analysis. The protective efficacy of subunit vaccine was assessed by ELISA which showed the highest protection (80%) against the virus challenge than that of commercial inactivated vaccine (70%).

Conclusion: The recombinant fiber-2 protein was determined to be a good option for a recombinant subunit vaccination to control HPS. 

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Correlation of SNR Value on DPOAE Examination with HSP70 Levels in Blood and HSP70 Expression in Cochlea of Noise Model Rattus norvegicus as an Indicator of Inner Ear Damage
Florensia Elita Pratiwi, Tengku Siti Hajar Haryuna, Indri Adriztina, Khalisanni Khalid, pages 322-328

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 Background: Cellular stress caused by noise-induced hearing loss can be observed through the damage of cochlear hair cells, mechanically and metabolically. SNR value on the DPOAE examination assesses the sensory function of cochlear outer hair cells. HSP70 as marker of cellular stress can be identified from its levels in blood and its expression in the cochlea.

Methods: Three groups of rats were used for the noise intervention: Group 1 was the control group, Group 2 had a noise exposure of 100 dB, and Group 3 received a noise exposure of 110 dB. DPOAE examination was then conducted, blood samples from rats’ tail were collected after a noise treatment, followed by the measurement of HSP70 levels by ELISA readings. Rats were terminated afterwards and immunohistochemical examination of the cochlear organ of Corti was performed to calculate the expression of HSP70. The data obtained were analyzed using SPSS.

Results: A decrease occurred in the values of SNR and an increase occurred in HSP70 levels in blood along with its expression in the cochlea of noise model Rattus norvegicus, and there is a correlation between the three.

Conclusion: The decrease in SNR values, increase in HSP70 levels in blood, and increase in HSP70 expression in the cochlear organ of Corti of Rattus norvegicus, proves that noise-induced hearing loss triggers the production of HSP70 as a cytoprotective agent in preventing inner ear damage. 

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Gene Expression Profiling to Predict Prognostic Biomarkers for Relapse in Multiple Myeloma
Sana Elahi, Sahar Fazal, pages 329-337

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 Background: Multiple myeloma (MM), an incurable malignancy of plasma cells (PCs), is the second most common hematological cancer caused primarily by structural variations (chromosomal aberrations and somatic mutations). The majority of the patients acquired resistance against standard therapeutic approaches for MM and experienced relapse despite the continuous advances in MM therapies.

Methods: We performed Differential Gene Expression (DGE), literature mining and SNV analysis of Next Generation Sequencing (NGS) data of Newly Diagnosed MM (NDMM) and Relapsed/Refractory MM (RRMM). The selected Differentially Expressed Genes (DEGs) were subjected to functional enrichment and pathway analysis. Immune cells infiltration analysis was also performed to estimate immune cells variations in the Tumor Microenvironment (TME) of RRMM. 

Result: CSF1R, VCAN, NRP1, COL22A1, BPI, BIRC5, MNX1, FAT1, ERG, TCL1A, AFF3 were selected after DGE, literature mining and SNV analysis. The functional enrichment of these DEGs showed significant enrichment for positive regulation of cell population proliferation, serene/threonine kinase activity, endothelial cell proliferation, cytokine binding, G protein activity and GDP binding, whereas KEGG pathway analysis revealed vital role of PI3K-Akt signaling pathway along with various cancer pathways. The immune cells infiltration analysis revealed the higher count of neutrophils and  lesser level of T cells (CD8+) in TME of RRMM. 

Conclusion: Our study suggests that neutrophils play an important role in modulation of TME in RRMM. The selected DEGs have previously been identified in progression, drug resistance and relapse of various cancers. The role of these biomarkers in RRMM has not been explored yet. Therefore, neutrophils, selected DEGs and PI3K-Akt signaling pathway are potential targets to investigate in RRMM. 

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In-vitro study on antioxidant status of Zinc NPs and crude extract of Ayurvedic herbal formulation
Jaafar Faez Kadhim Alsadooni, Sura Razzaq Khudhair, pages 338-345

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 Background: To create and evaluate cutting-edge therapeutic composition on biosynthesized nanoparticles exhibiting varied biological activity, nanotechnology is a novel and forward-thinking method. Increased antioxidant activity decreased toxicity to free radicals as well as cancer cells, and other benefits are achieved carefully by regulating the form and nanoparticle dimensions.

Methods: Nanostructures biosynthesized utilizing Liv-Pro-08 AHF showed a strong antioxidant effect. The creation of novel and more efficient antioxidants may benefit from the usage of nanoparticles. It is noteworthy that the resulting nanoparticles exhibit more biological processes than the extract. This research aims to calculate if there are any noticeable differences in the behaviour of leaf extracts and the many nanomaterials obtained from the aqueous-based separation of Liv-Pro-08 Ayurveda herbal-based formulations.

Results: Several in-vitro free radical scavenging experiments were used to determine whether or not Liv-pro-08 nanoparticles possessed any antioxidant properties. Zinc nanoparticles generated from Liv-pro-08 ayurvedic formulation demonstrated dose-dependent suppression of DPPH, as well as concentration-dependent lowering power potential, indicating that the nanomaterials contain free radical scavenging capability to produce the antioxidant effect.

Conclusion: Nanoparticles might be useful for the development of newer and more potent antioxidants. It is worth mentioning that the resultant nanoparticles possess an elevated biological activity in comparison to the extract. The data represented in our study contribute to a novel and unexplored area of nano materials as alternative medicine. 

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Isolation and Purification Glycyrrhizic acid and Determination of the Biological Activate Against Microbial Pathogens
Ayat Abdaljabar Al-Kubaise, Ayyad W. Al-Shahwany, pages 346-353

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 Background: The World Health Organization estimates that 80% of the world's population uses plant extracts or their active components in traditional therapies. Recently, there has been a lot of interest in the quest for new antioxidant and antibacterial plant-based medications. Additionally, Glycyrrhiza glabra is a popular food flavoring and sweetening ingredient.

Methods: This work was to isolate and purify licorice root's glycyrrhizin acid (GA) and investigate its antimicrobial properties against S. aureus and K.  pneumoniae were isolated from urine , blood, wounds, ear water, sewage, soil, and water, totaling 12 isolates of each kind, Analytical method for the measurement of Glycyrrhizic acid  from the root extract that has been suitably supported by a mass spectroscopic analysis.

Results: The result showed the effects of GA with different concentrations on S. aureus and. K. pneumoniae. Obviously, the MIC range value was in concentration between 18.75 to 37.5 % for S. aureus isolates, and 4.68 to 75 for K. pneumoniae isolates. The result indicated that GA reduced the metabolic activity of cells in biofilm on S. aureus and K. pneumoniae with an inhibition percentage

Conclusion: GA may have an effect on cell membrane permeability, biofilm formation, and efflux activity, which inhibits bacterial growth. 

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Exploring Genomic Patterns to Identify Drug-Resistant TB: A Comprehensive Study of Age, Gender, Lineage, and Outcome
Anam Tariq, Sahar Fazal, pages 354-361

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 Background: Tuberculosis (TB) is a prevalent bacterial infection caused by Mycobacterium tuberculosis (Mtb), primarily affecting the lungs, with rising incidence rates. Drug-resistant TB strains, including multidrug-resistant (MDR) and extensively drug-resistant (XDR) types, pose challenges with poor treatment success rates and increased mortality. Demographic factors and genomic patterns contribute to drug resistance, emphasizing the need for new drugs targeting specific genomic patterns.

Methods: This study analyzes the impact of demographics, treatment outcomes, and genomic mutations on drug-resistant TB using pattern identification techniques and whole-genome sequencing to discover therapeutic targets. This study primarily focused on the identification and analysis of MDR and XDR strains, considering various demographic characteristics and treatment outcomes. A dataset comprising 2,602 observations was utilized, and pattern recognition techniques were employed to identify significant features. Additionally, the study employs the whole-genome sequencing (WGS) pipeline to identify single nucleotide polymorphisms (SNPs) that result in new mutations in drug-resistant Mtb strains.

Results: The findings of the study revealed that XDR and MDR non-XDR TB were the most prevalent types of drug resistance, and they were associated with unfavorable treatment outcomes, including death or treatment failure. Males exhibited a higher susceptibility to both XDR and MDR non-XDR TB compared to females. The age of onset for both types of resistance was approximately 40 years. Among the observed variants in 31 commonly occurring genes,

Conclusion: These findings introduce a novel set of therapeutic targets specific to MDR and XDR Mtb types, which warrant further investigation for potential therapeutic interventions. 

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Immobilization of lipase enzyme extracted from thermophilic Bacillus licheniformis 14T local isolate
Saad Hussein Khudhair, Melad Khalaf Mohammed, Ahmed Darweesh Jabbar, pages 362-367

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 Background: Currently, lipase enzymes are considered important bio-catalysts in many industries due to their unique properties in catalyzing various types of reactions in aqueous solutions.  By targeting hydrocarbons in the oil, lipase enzymes contribute to the breakdown of hydrocarbons, reducing the environmental impact of oil spills and facilitating the remediation of contaminated areas.

Methods: A thermostable lipase from local isolate Bacillus licheniformis 14T has been immobilized on four different supports that include the inactivated chitosan beads, activated chitosan beads with glutaraldehyde, inactivated chitosan-alginate beads, and activated chitosan-alginate beads with glutaraldehyde.

Results The purified free lipase enzyme exhibited the highest enzymatic activity at 34.6 units/ml, surpassing all immobilized enzymes. Specific activity increased to 96.25 and 79.03 unit/mg protein for activated chitosan beads and activated chitosan-alginate beads, while decreasing to 52.95 and 46.05 unit/mg protein for inactivated chitosan beads and inactivated chitosan-alginate beads compared to the free enzyme. Optimal conditions for the immobilized enzyme differed, with the highest enzyme activity achieved after 60 minutes at 60°C and pH 8, reaching 48.6, 70.23, 43.12, and 61.2 units/ml on various supports, contrasting with the free enzyme's peak activity after 30 minutes at 50°C and pH 7.

Conclusions: Immobilizing the lipase enzyme increases the specific activity of the immobilized enzyme on the supports of activated chitosan beads with, also the immobilization process led to a change in the optimal conditions for the activity of the immobilized enzyme compared with the optimal conditions of free enzyme. 

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Microbial Contamination and Antibiotic Resistance in Food and Water: Assessing the Threat of Staphylococcus aureus in Lahore Metropolitan
Gulnaz Pervaiz, Zahoor Qadir Samra, Amina Hussain, Aroosha Hussain, Fareeha Javaid, pages 368-374

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 Background: The research project in Lahore, Pakistan, seeks to detect S. aureus in drinking water, raw milk, and yogurt samples due to health concerns. S. aureus is a pathogenic bacterium with potential risks if present in food and water sources.

Method: In this study, 300 samples of raw milk, yogurt, and drinking water were collected in Lahore. The presence of S. aureus was determined through morphological, microscopic, and biochemical methods. The biochemical analysis included testing for specific features of S. aureus. Disk diffusion technology was employed to assess the antimicrobial susceptibility of the isolates, following the recommendations of the Clinical Laboratory Standards Institute. Molecular confirmation was achieved through 16S rRNA sequence analysis using universal and specific primers.

Results: The investigation uncovered that 6% of drinking water samples, 9% of yogurt samples and 58% of raw milk samples were tainted with S. aureus. These findings were further validated through 16S rRNA sequence analysis, affirming their reliability. S. aureus exhibited notable resistance rates, with 100% resistance to penicillin and 95% resistance to erythromycin. Conversely, resistance to ciprofloxacin and gentamicin was lower, at 10% and 5% each, indicating the potential efficacy of these antibiotics in treating S. aureus infections.

Conclusion: The study emphasizes the risk of S. aureus infection from raw milk consumption in Lahore, Pakistan, due to inadequate sanitary practices. It stresses the necessity of implementing stricter measures in dairy production and water treatment to ensure public safety and reduce multidrug-resistant bacteria prevalence. Continuous monitoring and preventive actions are vital for safeguarding public health. 

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Hematological effects for rabbits immunized by Corynebacterium pseudotuberculosis sonicated antigen
Basil R. F. Razook, Mariam Hamdi Abdulkareem, Ansam Khalid Mohammed, pages 375-379

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 Background: Analyzing Hematological parameters is usually in use to monitor various circumstances such as infection, inflammation and anemia. For that we studied blood parameters which are in touch with immunized laboratory animals (rabbits) after immunizing these animals with Corynebacterium pseudotuberculosis then used an adjuvant of Pseudomonas aeruginosa to stimulate the immune response cells after 2 to 3 weeks of immunization. Different concentrations were used to examine the effect on the animals’ blood parameters changing.

Methods: Corynebacterium pseudotuberculosis bacterium was used at current study for evaluating the effect of immunizing laboratory rabbits with two different immune stimulators working together as adjuvant on the blood picture with the aid of 12 breed rabbits from different genders, dividing them at the base of inoculation with pre-killed and sonicated cells of bacteria (antigens) onto 4 categories as following: the group-1 members of rabbits were injected with the antigen, group-2 and group-3 included rabbits injected with the two inoculations at various concentrations; final group, group-4 was the control group.

Results: Thus, white blood cells known to be our main line of defenses act firstly in our body towards different infecting microorganisms, their count always was increased through the immunization correlated with other measured parameters. From the results we can see that the lymphocytes percentage had no alteration between the infected and the immunized compared to control group, while the percentage of MID that refers to the monocytes was at the same range in both infected and the immunized control rabbit’s groups. Granulocyte cells percentage, which is including heterophilic, basophilic and eosinophilic cells, was significantly decreased (P>0.05) compared with control at all groups.

Conclusion: We conclude that Cpseudotuberculosis might function as a potent immunogen to notice the complete blood picture variations. 

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Evaluation of the effectiveness of coniferous-phytogenic feed additives in case of eimeriosis infestation in calves
Vasily Korotkiy, Olga Skornyakova, Victoria Leukhina, Evgeny Sadykov, Viktor Ryzhov, pages 380-385
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 Background: Eimeriosis in calves, often treated with harsh chemicals, leads to significant health and economic burdens. This study explores natural coniferous-phytogenic feed additives as an innovative solution, focusing on their efficacy against Eimeria bovis and Eimeria ellipsoidalis, two common eimeriosis pathogens in calves.

Methods: The authors conducted experiments to analyze the therapeutic and preventive effectiveness of the use of a coniferous-phytogenic immunomodulator in the diet of calves from 5-7 days of age and a coniferous and salicylic acid feed additive from 28-30 days of age in case of eimeriosis infestation at a dose of 5.0, 10.0, and 15.0 ml, once a day, in courses of 2 weeks without intervals with a small amount of warm water.

Results: The extensivity of the coniferous and salicylic acid feed additive against the eimeriosis infestation in calves caused by E. bovis showed maximum result. Both preparations were willingly consumed by calves and eliminated the signs of diarrheal dyspeptic syndrome and gastroenteritis. The inclusion of coniferous-phytogenic feed additives in the calves' diet from 5-7 days of age once a day, in courses of 2 weeks without intervals, contributed to 100% effectiveness against E. bovis and E. ellipsoidalis.

Conclusion: The study concludes that coniferous-phytogenic feed additives are highly effective in treating and preventing eimeriosis in calves, showing 100% effectiveness against the targeted pathogens. These additives also contribute to the overall health and well-being of the animals by alleviating related symptoms.

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Histopathological Detection of the Protective Role of Hydroxytyrosol Against the adverse Effect of Azithromycin in Rats
Ghusoon Abdul Kareem Neamah, Ekhlas Abid Hamza Alalwany, Fadak Bahaa Rabee, Adnan Mansour Jasim, Ahmed Samir Abukhomra, pages 386-391
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 Background: Adverse medication reactions affecting one or more organ systems can manifest as side effects of antibiotics. There is a possibility that some antibiotics will have adverse effects that are fatal. The present study aims to examine the ability of  hydroxytyrosol (HT) to protect the liver,  heart , and improvement of kidney function against the adverse effects of azithromycin.

Methods: Thirty adult male rats were randomly divided into three equal groups of 10 animals each. The first group, designated negative control (NC), was the daily oral administration of normal saline. The second group (T1) was subject to daily oral administration of azithromycin (30 mg/kg). The last third group (T2) was administered azithromycin (30 mg/kg) with hydroxytyrosol (50 mg/kg). After two weeks the animals were sacrificed for serum collection, histopathological specimens, and drug docking evaluation using Auto Dock Vina.

Result: The present study showed that score binding in 18gs Glutathione S-transferase was preferable in hydroxytyrosol with (-5.95 Kcal/mol), while vitamin E was in (-7.80 Kcal/mol) and (-6.16 Kcal/mol) for glutathione. On the other aspect, antioxidant coenzyme Q10 showed less binding with 18gs Glutathione S-transferase to recorded binding at (-4.9) with Root Mean Square Deviation (RMSD) (29.3, 30.3, 23.9 Kcal/mol) and (28.6) Kcal/mol, respectively. The histopathological result for T1 showed  several histological effects in the liver, heart, and kidney, while in T2 group showed improvement in tissue architecture. 

Conclusion: The study concluded that hydroxytyrosol binding glutathione S-transferase could predict potent antioxidants near glutathione, might be responsible for scavenger free radicals, and protect the tissue against the toxic effects of azithromycin. 

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Antifungal Efficacy of the crude Alkaloid, Flavonoid, and Terpenoid of Saussurea costus (Falc.) Lipschitz Roots against Aspergillus species isolated from Rice Seeds
Fatima S. Al-Nafie, Hussein J. Hussein, Abeer Fauzi Al-Rubaye, pages 392-397
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 Background: Rice is an essential component of the diets of people all over the world, and it accounts for a sizeable amount of the calories that they consume. The world's food supply suffers huge economic losses due to fungal deterioration on rice seeds. The medicinal plant Saussurea costus contains bioactive chemicals and is well-known for its many therapeutic uses. There are a number of species of Aspergillus that are considered to be diseases in vegetables and fruits. One of these species is Aspergillus niger. The purpose of this current investigation was to investigate the impact of the crude Alkaloid, Flavonoid, and Terpenoid chemicals derived from Saussurea costus' roots on Aspergillus species Purified from Rice seeds obtained from various markets in Hillah, Iraq.

Methods: In vitro, antifungal activity against Aspergillus species was achieved using the food poisoning method. Three concentrations 5, 10, and 15mg/ml of each crude compound were prepared and compared to a positive control represented by Carbendazim500g/l and a negative control represented by 10% dimethyl sulfoxide.

Results: An objective of the current investigation was to manage the Aspergillus species growth identified in Rice seeds by utilizing secondary compounds derived from the roots of Saussurea costus. The study's data indicated that the extracts represented by active compounds such as Alkaloids, Flavonoids, and Terpenoids obtained from Saussurea costus roots exhibited a significant decrease in the of Aspergillus species growth, particularly at a concentration of 100mg/ml, in contrast to the negative control. This effect was equivalent to that of the positive control, Carbendazim 500g/l.

Conclusion: Finally, Saussurea costus roots have shown the most efficacy in controlling Aspergillus species, particularly through the presence of Alkaloid, Flavonoid, and Terpenoid chemicals at a concentration of 100mg/ml. 

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First report of Tomato leaf curl New Delhi virus (a bipartite begomovirus) from round gourd plant in Swat region of Pakistan: its phylogenetic and biogeographic analysis
Chand Bibi, Raham Sher Khan, Murad Ali Rahat, Asadullah, Muhammad Zahoor, Hassan Sher, Khadim Hussain, Fazal Akbar, Mohammed Ali Al-Saleh, pages 398-405
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 Background: The family Geminiviridae contains plant viruses commonly referred to as geminiviruses, which cause diseases in numerous important plants. Geminiviridae is one of the most important family of plant-infecting viruses that has been divided into fourteen recognized genera.  Amongst these, the genus Begomovirus is the largest and economically most important one, viruses of the genus are further categorized into bipartite (two genomic components called DNA-A and DNA-B) and monopartite (single genomic component or DNA-A) begomoviruses.

Method: The genomic DNA was extracted from symptomatic plant samples and both the DNA-(B and A) components of a bipartite begomovirus were amplified via rolling circular amplification (RCA) and PCR. The amplified DNA-A and DNA-B were cloned and sequenced by Primer walking, Sanger sequencing, and Illumina sequencing methods. The phylogenetic and biogeographic analyses of both components were performed by various bioinformatics tools.

Results: DNA-A (accession number MW722701) and DNA-B (accession number MW722782) of begomoviruses that were sequenced shared >95% sequence similarity with the bipartite begomovirus known as Tomato Leaf Curl New Delhi virus (TLCuNDV). Phylogenetic and biogeographic analyses proposed several possible ancestors, and multiple dispersal and vicariance events may be involved in the evolution of this virus. 

Conclusions: This is the first time a bipartite begomovirus has been found to infect a plant in the study area (District Swat, Pakistan). The existence of ToLCNDV may pose a major danger to other important crops in the region. 

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Effects of Acetaminophen and Vitamin C Combination on Clinical Vital Signs and Behavioral Traits of Heat-Stressed Dairy Buffalo
Wisam K. Salih Sangor, Adnan Jabbar J. Al-Kanaan, Nameer A. Khudhair, pages 406-413
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 Background: Heat stress (HS) presents significant challenges to water buffalo breeding, particularly in harsh environments like southern Iraq. HS adversely impacts physiological and behavioral aspects in dairy buffaloes, leading to economic losses in productivity and welfare. Interventions such as acetaminophen and vitamin C supplementation have been suggested to alleviate HS impacts due to their anti-stress and anti-inflammatory characteristics.

Methods: This study was conducted in Al-Chibayish Marshes southern Iraq and aimed to evaluate the effects of acetaminophen and vitamin C combination (AVC) on heat-stressed dairy buffalo. Sixteen lactating buffaloes were divided into four treatments. One treatment received regular water (control), while other treatments were given water mixed with AVC at a dose of 500g/1000 liters for 3, 5, and 7 consecutive days per week, respectively, over 30 days. Environmental (temperature, humidity, and THI) and physiological data (body temperature, respiration rate, and pulse rate) were recorded daily, while feed and water consumption were recorded weekly.

Results: Buffaloes experienced significant HS, with temperatures reaching up to 53.2°C and corresponding THI values ranging from 88.05 to 99.31. AVC administration for 5 consecutive days led to increased feed intake by 1.15 kg daily per treatment and decreased water consumption by 16.43 liters daily per treatment compared to the control. Additionally, rectal temperature, respiration rate, and heart rate showed progressive improvement during the second and third weeks, with the most notable improvement observed in the fourth week of the study. These improvements in vital signs and behavioral traits indicate the ability of AVC to improve thermoregulation and reduce heat stress impact.

Conclusion: The investigation highlights the potential of AVC supplementation showing a promise in alleviating HS effects on dairy buffaloes by positively influencing physiological and behavioral parameters. Further research is needed to elucidate the underlying mechanisms and optimize supplementation strategies for enhancing buffalo welfare and productivity under heat stress. 

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The effect of inflammatory cytokines on occurrence of retained placenta in cattle
Nawaf Nooraldeen Dhaher, Manar Sabah Eesa, Hayder A. H. Al-Mutar, Maythem Abdulealah Ismaeel, pages 414-418
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 Background: In simple terms, retained placenta is a common issue after parturition in cattle that can affect their reproductive ability. In pregnancies with fetal growth restriction (FGR), the placenta has low anti-inflammatory cytokines and high pro-inflammatory cytokines. By looking at variations in cytokine levels in the blood, we can diagnose the condition. The focus of a recent study was to examine the role of some interleukins  (Interleukins -10, Interleukins -6, Interleukins -1β), C-reactive protein (CRP) and Tumor Necrosis Factor (TNF)-α, in retained placenta occurrence in cattle.

Methods: The study involved 40 cows, aged 3-8 years in the Salah-Din province. Each animal suffered from retained placenta also eleven healthy cows served as the control group in a period of January to August 2022. The blood serum was analyzed using enzyme immunoassay techniques (ELIZA) to measure the levels of IL-10, IL-6, IL-1β, CRP and TNF-α.

Results: Serum levels of IL-10, IL-6, IL-1β, CRP and TNF-α showed a significant increase in cows experiencing retained fetal membranes compared to healthy cows (2.31±0.11 vs 1.41 ±0.07) (10.48±0.24 vs 5.40±0.19), (13.6±2.1 vs 4±0.9), (0.9±0.02 vs 0.32±0.04) and (60.1 ± 12.79 vs 29.5 ± 16.58) respectively.

Conclusion: from the present study we conclude that IL10, IL6, IL1β and TNFα play an essential component in retained placenta incidence, and  the estimation levels of these parameters in serum may be considered good indicator for occurrence in cattle. RT-PCR revealed increased expression of SDHA genes in the maternal compartment of the placenta.  

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Association between vitamin D levels and insulin resistance in patients with type 2 diabetes mellitus
Muntaha Abdulmajeed Arif, Manal Kamal Rasheed, Ammar Adil Ismaeel, pages 419-424
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 Background: Insulin resistance (IR) is a major contributor to type 2 diabetes mellitus (T2DM) and is associated with various metabolic disorders. Vitamin D deficiency (VDD) has been linked to an increased risk of T2DM, but the relationship between vitamin D status and IR in T2DM patients remains unclear. This study explores this association in a male population with T2DM in Baghdad, Iraq.

Methods: A case-control study was conducted on 190 male participants: 40 healthy controls and 150 patients categorized into different groups; 40 diabetic patients, 55 with coronary artery disease (CAD) and T2DM (CAD+DM), and 55 with only CAD. Physicians diagnosed them, and they were evaluated by physical and full medical history. All the sample set parameters were measured in the fasting state for each group. The biochemical tests included body mass index (BMI), vitamin D levels, insulin levels, fasting tests glucose (FBG), and HbA1c were measured. Insulin resistance was calculated using the HOMA-IR model.

Results: Vitamin D levels were significantly lower in all diabetic groups (DM, CAD+DM) compared to the control group (p<0.001). All diabetic patients had insulin resistance. BMI was significantly higher in the CAD and CAD+DM groups compared to controls (p<0.00001, p<0.004). FBG, HbA1c, insulin levels, and HOMA-IR were all significantly higher in diabetic patients compared to controls (p<0.001 for all). Furthermore, increased HbA1c, fasting insulin, and homeostatic model assessment for insulin resistance (HOMA-IR) were associated with CAD progression in T2DM patients (p < 0.001).

Conclusion: This study suggests that vitamin D deficiency may contribute to insulin resistance in male T2DM patients in Baghdad, Iraq. Maintaining optimal vitamin D levels may be beneficial in diabetes management in this population. Incorporating vitamin D screening into routine check-ups for T2DM patients could aid in early detection and prevention of insulin resistance-associated complications. 

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Changes of Serum Midkine as a Dynamic Prognostic Factor in Detection for Thyroid Cancer and Comparison with Fine-Needle Aspiration
Hadel Kareem Al-Rubaiawi, Raid J. Mohamed, Sajid H. Alhelfy,  Marwah S. Yones, pages 425-429
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 Background: Midkine plays a significant role in the pathophysiology of malignant and other diseases since it is involved in a number of physiological processes, including development, reproduction, and repair. Different types of cells express this protein. Thus, in healthy conditions, significant MDK expression is observed in lymphocytes, the epidermis, and the bronchial epithelium. The sample obtained during fine needle aspiration can help in the diagnosis of or rule out disorders like cancer. Blood is the most practical and least likely bodily fluid to pollute while diagnosing illnesses. Finding successful prognostic markers in patients with well-defined thyroid cancer was the goal of the current investigation.

Methods: Midkine level was measured in serum samples by using Elisa technique for 80 patients (40 patient was diagnosed with papillary thyroid carcinoma and another 40 with benign thyroid goiter), Age (25-55) from Iraqis people.

Results: The current study revealed highly significant difference the median serum level of medkine in the TC patients was much higher than that of BTT patients with a highly significant difference The sensitivity and specificity of the test at cut off value of medkine=129 pg/ml were 90% for both. While FNA result, the sensitivity and specificity of FNAC was 80.77% and 89.66% respectively.

Conclusions: The thyroid cancer group's serum MDK result was noticeably higher than that of the healthy control groups. proposed that the midkine might serve as a biochemical marker for thyroid cancer early identification and diagnosis. 

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Acquiring insights through a sequence-based approach to the critical Zika virus MTase domain
Banan Atwah, Saad Alghamdi, Nizar H. Saeedi, Abdulrahman Alzahrani, Rashed Mohammed Alghamdi, Asif Hussain Akber, Mohammed Yahya Al Qahtani, Atiah Abkar Yahya Mujarribi, Saeed Saleh Al Qahtani, Ahmed Mohammed Faqihi, Mohammad Azhar Kamal, pages 430-437
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 Background: ZIKV is one of the re-emerging arboviruses (viruses carried by arthropods), which is spread through the Aedes mosquito. It is an RNA virus with only one strand that is appropriate to the family Flaviviridae's  Flavivirus (genus) & has been linked to other Flaviviruses such as the West Nile virus, chikungunya virus, & dengue (DENV) virus. The envelope, precursor membrane, and capsid are three structural proteins, and seven nonstructural proteins are also encoded by the Zika virus genome.

Methods: We conducted an in-silico analysis of the Zika virus' MTase domain protein for this publication. We predicted that methylation would play a significant role in the available Prosite, Pfam, and InterProScan tools to aid in locating the MTase domain. Along with alignment, amino acid composition, charged amino acids, atomic level studies, & molecular weight, we also make predictions for these variables, including theoretical Pi.

Results: We also examine the MTase domain's simulated structure (alpha helix, beta sheet, turn) and its specifics, including secondary structure. We also pinpoint the locations where proteins, DNA, and RNA bind. Potential phosphorylation sites can be found on the Ser, Thr, and Tyr residues in the MTase domain.

Conclusion: These outcomes imply a complicated interaction between different phosphorylation modifications that modulates the activity of the MTase domain. To fully appreciate the auxiliary and practical perspectives and to clarify the varied roles of PTM in the MTase domain will be a primary goal of future study. 

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Whole genome sequence of Lactiplantibacillus plantarum strain HA9 isolated from conventional Iraqi cheese: First report
Hasanain Qasim Mezaal Al-kaabi, Rahman Laibi Chelab, pages 182-187
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 Background: Lactiplantibacillus plantarum is a renowned probiotic known for its ability to enhance health when ingested in sufficient quantities. While L. plantarum typically exhibits unique characteristics, variations in strains and environments can occur. Therefore, it is crucial to conduct genetic identification of the strain before incorporating it into products or administering it for health benefits.

Methods: After being isolated from classic artisanal milk cheeses, the bacterial DNA extraction kit from GENEAID, Korea, was used, and strain HA9's genomic DNA was isolated. Then, the highly conserved section of ribosomal RNA was amplified with universal primers. The results were confirmed by a blast analysis of the NCBI database. The genome of HA9 was sequenced using the Illumina and PacBio Sequel II technologies (HN00194138; Psomagen/USA). The Prokaryotic Dynamic Programming Gene-Finding Algorithm software was applied to forecast coding genes. The Pathosystems Resource Integration Center constructed the L. plantarum genome.

Result: According to the findings, the genome of the L. plantarum HA9 strain has a length of 3,321,986 bp and a GC percentage of 44.40. This assembled genome contained a total of 1,420 coding proteins and 82 contigs, as discovered by the Comprehensive Genome Analysis. The isolate had been submitted to the NCBI and received the HA9 genome sequence (accession ID: JAUTDL000000000.1).

Conclusion Microbes' whole-genome sequencing plays a crucial role in clarifying their probiotic properties and enables accurate identification down to the strain level. The authors argue that further research should be done on probiotics' whole genomes, with a special emphasis on second-generation strains. 

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Bacteriophage Endolysins Antibacterial Effect against S. mutans Isolated from Dental Caries: An In-Vitro Study
Huda Mohammad Hasan Ali, Adeeb Ahmed kadhim Al-Zubaidy, Ahmed Sahib Abdulamir, pages 444-448
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 Background: Many bacterial infections including dental caries that caused by Streptococcus mutans (S. mutans) were difficult to be treated due to the unavailability of effective antibiotics. Therefore, it requires to focus the attention on endolysin as an antibacterial agent. This study was designed to evaluate the In-vitro  effectiveness of specific bacteriophage endolysins against S. mutans isolated from dental caries.

Methods: Twenty-five isolates of S. mutans were collected from patients with dental caries attended to Al-Imamain Al-Kadhimain, Medical City Hospital, and AL-Ameen Health Center, during the period from December 2021 to May 2022. The S. mutans isolates were identified by culturing them on their specific mitis salivarius sucrose bacitracin tellurite media (MS-SBT agar), morphological characteristics, gram staining, biochemical tests and VITEK 2 compact system. Seven of these S. mutans isolates were used for the in-vitro study.

Results: The bacteriophage endolysin enzyme extracted from the corresponding S. mutans isolate was effective as In-vitro  antibacterial agent with minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) ranging between (13.63 – 17.83) µg/ml and (17.04-22.28) µg/ml respectively.

Conclusion: The endolysin was effective as antibacterial agent against multi-drug resistant (MDR) S. mutans upon growing and testing on culture media. 

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In Silico Identification of Novel HDAC2 Inhibitors for Reinstating Synaptic wiring in Autism Spectrum Disorder
Samir Abdulkarim Alharbi, pages 449-454
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 Background: Autism spectrum disorder (ASD) is a neurodevelopmental disorder with an increasing incidence rate. For the treatment of ASD, several pharmaceutical approaches, dietary supplements, and behavioral therapy have been proposed, but a definitive cure remains elusive. Histone deacetylases (HDACs) are critical epigenetic regulators whose molecular and pharmacological roles are being studied extensively in medically significant ailments such as neuropsychiatric disorders, neurodegenerative diseases, and cancer on a global scale. HDAC2's specific expression pattern in CNS makes it an appealing therapeutic target for neurological illnesses such as ASD.

Methods: The study used PyRx software (version 0.8) to screen a library of 401 alkaloid compounds against HDAC2. In addition, the DS software was used to predict the physicochemical and DMET properties of the compound library.

Results: Valaciclovir hydrochloride, Dihydrocapsaicin, Guanosine, Santacruzamate A, and 2'-Deoxyguanosine monohydrate compounds exhibited strong interactions with HDAC2. These compounds had a higher binding energy than the control compound, and they have promising drug-like properties and ADMET characteristics.

Conclusion: These identified compounds could be used as HDAC2 inhibitors for the management of ASD, however, experimental research is needed to optimize them as HDAC2 inhibitors. 

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Assessing the Diagnostic Precision of Chosen Equational Scores in Detecting Steatosis through Elastography
Sajid Alhaidari, B Charfeddine, Talal Hadi, pages 455-461
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 Background: Several non-interventional imaging studies, scoring systems, and biochemical markers are widely used nowadays to urgently diagnose stage and manage cases of non-alcoholic fatty liver disease (NAFLD) to avoid potential complications. The current study aimed to compare the sensitivity, specificity, and positive predictive values (PPR) of some widely used NAFLD-induced steatosis scoring systems.

Methods: Fifty patients confirmed with fatty liver by ultrasonography were recruited in this study. All patients underwent steatosis staging using FibroScan. For all patients, aspartate transaminase (AST), alanine transaminase (ALT), platelet count, and body mass index were measured for the calculation of aspartate transaminase to platelet ratio index (APRI), hepatic steatosis index (HSI), and AST/ALT equations. Sensitivity, specificity, and PPR for the three scoring systems were calculated compared to FibroScan scoring results as the standard validating system where S0 and S1 scores were considered negative while S2 and S3 scores were considered positive for hepatic steatosis.

Results: Sensitivity results were 45, 87.8, and 45.9%; specificity results were 30, 22.2, and 69.2%; and PPR results were 72, 83.7, and 82.9% for APRI, HSI, and AST/ALT ratio scoring equation, respectively.

Conclusion: Among the three steatosis scoring systems studied in the present study, the HSI was the most sensitive scoring system, and the AST/ALT ratio was the most specific scoring system compared to the FibroScan as the standard steatosis scoring system. 

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Study the Teratogenic Effect of Antimony on The Histological Changes of the Liver in the Adult Rabbit (Oryctolagus cuniculus)
Ibtisam Khalaf Abd Ali, Saba Dawod Salman, Thekra Atta Ibrahim, Mohammed Nsaif Abbas, pages  462-469
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 Background: Heavy elements such as antimony greatly affect the environment and living organisms. Antimony is discharged into the environment by mining and industries that use it as pesticides and flame retardants. This activity can lead to environmental pollution, water and soil contamination. Antimony can also accumulate in living organisms and cause negative health effects, such as damage to the respiratory system and skin, and growth abnormalities of animals and plants.

Methods: The primary objective of this investigation was to explore the teratogenic impact of the antimony heavy metal on histological structure of the liver in adult rabbits (Oryctolagus cuniculus). The study included 21 adult white rabbits divided into several groups: the first one is the control group injected with physiological saline (0.09% NaCl), the other group injected with 20 mg/kg antimony, and the last injected with 30 mg/kg antimony over a 30-day period. Following this, postmortem procedures were conducted to extract and fix the liver organ, and tissue sections were prepared.

Result: The results revealed significant histological changes, including distortion and rupture in Glisson’s Capsule, leading to the formation of a sub-capsular space due to its separation from hepatocytes. Additionally, alterations in the radial organization of hepatocytes and pyknosis in the nuclei were observed, characterized by a dark color and reduced size. Karyolysis, where nuclei completely disappeared, and hydropic degeneration in hepatocytes with swollen appearance and dark nuclei due to fluid accumulation were noted. Moreover, an increase in Kupffer cells and blood congestion in the central vein, resulting in dilation compared to the control group, were observed.

Conclusion: Overall, the treatment with antimony at 20 and 30 mg/g doses for 30 days show profound teratogenic effects on the histological structure of the liver in adult rabbits. These effects are represented by the destruction of various parts of liver, in addition to changes in arrangement, and distortion and rupture of the cells. Furthermore, an increase in Kupffer cells and blood congestion were also recorded. 

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Simultaneous detection of phylogroups and ESBL genes in E. coli using Multiplex PCR
Bushra B. Patoli, Atif A. Patoli, Iram Nabi Begum, Zarafshan Majeed, pages 470-476
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 Background: Escherichia coli (E. coli) strains harbor various allelic versions of beta lactamase genes and their identification using conventional phenotypic tests is a tedious and time consuming task. In the present study, multiplex PCR is performed for the simulatanoues detection of E. coli phylogroups and extended-spectrum β-lactamase- (ESBL) genes.

Methods: A total of 128 E. coli isolates from urine samples were screened for antibiotic resistance and expression of ESBL activity using phenotypic and genotypic methods. Uniplex and multiplex PCRs were used to detect E. coli phylogroup detrminants and blaCTX-M-15blaOXA-1 and TEM1 genes. Chi Square test of independence was employed for evaluating significant levels at P value < 0.05. 

Results: Phylogroup B2 was detected as the predominant group (36%) followed by group D (30%), A (25%) and B1 (9%). The highest resistance was seen against nalidixic acid (100%) and the lowest against amoxicillin-clavulanic acid (55%). Significant P values were observed for resistance against cefotaxime and cefepime in the phylogroup B2 while resistance against cefoxitin, sulfamethoxazole and fosfomycin was significantly associated with group D. Combination disc diffusion test (CDDT) showed ESBL activity in 42% E. coli isolates. A significant association of blaCTX-M-15 gene was observed for phylogroup B2 (P = 0.007). Moreover, a combination genotype of blaCTX-M-15 and TEM1 was also found statistically prevalent in phylogroup B2 (P = 0.006).

Conclusion: The study highlights the alarming rise in antibiotic resistance and delineates B2 a predominant phylogoup with a high prevalence of blaCTX-M-15 and TEM1 genes in urinary E. coli isolates. 

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Healing of Wounds by use of Spider threads in comparison with healing by Tetracycline ointment, an Experimental Analytical Study
Najla Siddig Nasir, Ranya Hafez Abdelrahman, Monzer Abass Fadul, Fatima Fadul Al, Mohamed Osman Elamin Bushara, Omer Siddig Nasir, Hatim A. Natto, Hatim M. Badri, Wahaj A. Khan, Ahmed A. Osman, Nazik Mubarek Hassan, Muath Aldomini, pages 477-481
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 Background: Spiders are distributed worldwide except in Arctic and Antarctica. More than 45,700 species of spiders and 114 families have been identified. Spiders produce silk in order to capture their prey. reported that people of the Carpathian Mountains used spider webs from Atypus spiders as bandages. Experiments showed that silk from spider house is bio-degradable, non-antigenic and non-inflammatory. These are ideal properties for healing wounds. We aimed to identify the effect of spider threads on healing wounds and injuries compared to healing of wounds by application of tetracycline ointment.

Methods: This is an experimental analytical study in which we investigated ten participants tested in three groups. Group A participants were treated with tetracycline and group B were treated with spider threads. wooden rulers used to collect spider silk threads along with sterilizer, tissue, paper, bandages, and Tetracycline ointment. Silk was collected from four types of spiders: Cellar Spider, Salticidae Spider, Venatoria Hetropoda, and Galeodes Arabs.  These spiders are the mostly distributed in Sudan. Silk was collected by using wooden rulers from cracks and corners of walls at Alzaiem Alazhari University and then placed on clean, soft tissue paper.

Results: revealed that spider threads heal surface injuries without leaving scars in 90% of participants. The healing process with spider threads lasts between 8 -24 hours. The comparison between three tests showed that the percentage of wound healing was highest by 70% by using spider threads in comparison with tetracycline and self-healing.

Conclusion: It was observed that healing of injuries by using spider threads was 70% faster than healing by using tetracycline ointment. Self-healing was observed to be faster than healing by use of tetracycline ointment. 

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Enhancement of oil biodegradation by using the biosurfactant produced from local Bacillus subtilis isolate
Melad Khalaf Mohammed, Saad Hussein Khudhair, Ahmed Darweesh Jabbar, pages 482-487
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 Background: Oil contamination poses a significant threat to the global environment and has attracted widespread attention in recent years. Given its significance, further exploration of biodegradation options for this contamination has never been more crucial than it is today. Therefore, the objective of the study was to improve the biodegradation of oil by utilizing local bacterial isolates alongside a biosurfactant produced by a Bacillus subtilis isolate. Twenty-two bacterial isolates were collected from four samples of hydrocarbon-contaminated soil. These isolates underwent screening to assess their effectiveness in degrading crude oil using two distinct methods.

Methods: Soli samples were collected from two contaminated sites with oil pollutants in Baghdad city. Biodegradation ability was tested on liquid Bushnell Haas medium (BH), pH 7 supplemented with 1% of crude oil and then screened primarily using the 2,6-dichlorophenol indophenol method to determine the ability of isolates to degrade the crude oil. All isolates were identified morphologically and biochemically. Biosurfactants was extracted from Bacillus subtilis previously isolated.

Results: Isolate SCS1 has appeared the highest ability reaching 52.6% compared with other isolates, also, the results of secondary screening confirmed that isolate SCS1 has given the best biodegradation reaching 53.8%. The isolate SCS1 was identified as Pseudomonas aeruginosa, this isolate was used to study the effect of biosurfactant on crude oil biodegradation. Results exhibited a high biodegradation efficiency reaching 85.1% in culture broths supplemented with 100 mg/100. It was noted that the use of 150 and 200 mg of biosurfactant led to a decrease in the biodegradation of crude oil.

Conclusion: The use of biosurfactant led to an increase in the degradation rate by up to 61.7% of crude oil by Pseudomonas aeruginosa when using 25-100 mg of crude biosurfactant that produced by local isolate Bacillus subtilis. 

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Structure-Based Virtual Screening of Antiviral Compounds Targeting the Norovirus RdRp Protein
Leena S. Alqahtani, Ahmad Salah Alkathiri, Abdulrahman Alzahrani, Rashed Mohammed Alghamdi, Waad Abdulrahmman Alamri, Mohammad Azhar Kamal, Ahmed Hamdan Aloufi, Ali Saeed Alamri, Qamre Alam, pages 233-236
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 Background: Human noroviruses (NV) are the primary etiological organisms causing acute gastroenteritis around the world, causing severe morbidity and imposing a significant economic burden. The RNA-dependent RNA polymerase (RdRp) is essential for viral replication and could be a promising target for anti-NV therapeutics. Despite the discovery of a few NV RdRp inhibitors, the majority of these pharmaceuticals have demonstrated limited efficacy in inhibiting viral replication in cellular models.

Methods: In this study, computational screening of antiviral compounds was conducted targeting the NV RdRp protein. The assessment was based on binding poses and the key residues of RdRp involved in interactions with compounds.

Results: The compounds namely, Ribavirin, BMS806, Dihydromyricetin, R7935788, and LY2784544 were found to bind the RdRp protein with high affinity. Notably, these compounds displayed significantly lower binding affinities compared to the positive control, PPNDS. In addition, these compounds exhibited many RdRp protein binding residues that were also present in the PPNDS.

Conclusion: The results presented here suggest that these compounds have the potential to be used as inhibitors of NV RdRp in the development of antiviral medications. Nevertheless, due to the computational nature of this study, it is imperative to do experimental validation. 

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Geospatial Dynamics of SARS-CoV-2 Variants during the Fifth Wave of COVID-19 in Punjab, Pakistan
Muhammad Mubashar Beig, Muhammad Asad Ali, Hasnain Javed, Tahir Yaqub, Asim Khalid Mehmood, Muhammad Nawaz, Sohail Raza, Muhammad Adnan Ashraf, Muhammad Abu Bakr Shabbir, Nadia Mukhtar, Muhammad Imran Najeeb, Waqar Aziz, Adnan Mehmood, Hamza, Awais Masud, Rida Basharat, Aftab Ahmad Anjum, pages 493-499
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 Background: The study was conducted in the various districts of Punjab, Pakistan, to ascertain the incidence and epidemiology of SARS-CoV-2 variants circulating in the population during the fifth wave of COVID-19.

Method: A total of 9603 nasopharyngeal swab samples of suspected patients were collected from the different districts of Punjab from December 2021 to April 2022. In the BSL-3 facility, an auto-extractor (Uni-medica) was used to extract the RNA genome. Viral detection and quantification were performed using real-time reverse transcriptase PCR. Multiplex PCR was used to target different mutations of the spike protein in order to identify SARS-CoV-2 variants.

Results: 711 samples were found to be positive from a total of 9603. The Omicron variant of concern (VOC) was the predominant lineage of SARS-CoV-2 circulating at the time of sampling. Ninety-one percent proportion of COVID-19 was caused by the Omicron, followed by the wild variant (3.80%) and Delta (68.11%). Men were found to have a greater Omicron prevalence (47.96%) than women (42.05%). Furthermore, compared to older individuals (32.07%) and younger kids (10.55%), adults had the higher percentage of Omicron (47.39%).

Conclusion: This study brought attention to the Omicron variant's circulation in Punjab Province, Pakistan, during the COVID-19 fifth wave. 

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Assessment of Knowledge and Attitude of First Aid Management of Epistaxis among the General Population in Najran Region: A Cross-Sectional Study
Ali Abdullah Alshehri, Ibrahim Nasser Al Sulaiman, Shuruq Talea Asiri, Sarah Saeed Aldughar, Nisreen Oudah Alqarni, Mohamed Osama Nour, pages 500-507
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 Background: Nasal bleeding, epistaxis, is a common emergency. Few Saudi studies exist on epistaxis awareness and first aid management. We aimed to assess knowledge and attitude of adults in Najran, Saudi Arabia towards epistaxis first aid management, as well as their association with sociodemographic factors.

Methods: A cross-sectional web-based survey was carried out involving general adults of Najran, Saudi Arabia, using a snowball sampling method. A pretested questionnaire was utilized for data collection. Knowledge and attitude scores were categorized as good/positive or poor/negative.  Both descriptive and inferential statistics were carried out with odds ratio (OR), 95% confidence intervals (CIs), and logistic regression analysis to investigate factors associated with knowledge and attitude towards epistaxis first aid management using SPSS Software.

Results: The study included 518 participants with 58.9% experienced nosebleeds. Common causes were hypertension (24.5%) and nose injuries (23.7%). Factors as nasal dryness, blood thinner use, and tumors/nasal polyps were also identified. Around 60.2% believed first aid was necessary and 59.5% had good knowledge about epistaxis first aid management. Participants with university/higher education, employed in health sector, and with previous experience with nosebleed were more knowledgeable, more likely to have positive attitudes, and showed significantly higher ORs for knowledge and attitude scores compared to others.

Conclusion: The study showed fair knowledge and attitude of Najran adults towards epistaxis first aid management with sociodemographic variables showing varying associations. Educational initiatives are needed especially for those with lower education, non-health sectors, and not experienced nosebleeds, to improve public first aid knowledge and attitudes. 

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Identification of Novel STAT3 Dimerization Inhibitor Through Structure-Based Virtual Screening for Cancer Management
Akram Ahmed Aloqbi, pages 508-512
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 Background: Signal transducer and activator of transcription 3 (STAT3) is a transcription factor that controls cell proliferation, differentiation, angiogenesis, and immunological responses. In many human malignancies, abnormal STAT3 activation promotes tumor growth via oncogenic gene expression, resulting in tumor malignancy. Many drugs with clinically authorized analogues that are used as STAT3 inhibitors for cancer therapy have several drawbacks in terms of stability and toxicity.

Methods: This study used PyRx 0.8 tool to screen the Traditional Chinese Medicine (TCM) database of about 32,364 commercially available natural compounds in order to identify new STAT3 inhibitors. Physicochemical and ADME properties of selected compounds were estimated using Datawarrior and SwissADME.

Result: The top 20 compounds were initially chosen based on their strong binding affinities with STAT3. Lipinski and Vaber tools were used to filter the top 20 compounds, yielding the top 6 compounds. The compounds ZINC85542844, ZINC4098720, ZINC85543599, ZINC85593523, ZINC85593528, and ZINC85593537 were passed through these filters. These compounds were found to interact with active site STAT3 residues and have several amino acid interactions in common with the control compound (STX0119).

Conclusion: This study suggests that the compounds ZINC85542844, ZINC4098720, ZINC85543599, ZINC85593523, ZINC85593528, and ZINC85593537 could be used as a lead for the development of novel STAT3 inhibitors. However, further experimental validation is required to optimized them as STAT3 inhibitors. 

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Alleviation of heat stress by ACC Deaminase and Exopolysaccharide Producing Plant-Growth-Promoting Rhizobacteria in Solanum melongena L.
Sami Hannachi, Karim Naghmouchi, Angelo Signore, pages 513-524
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 Background: The current study was carried out to assess the alternative role of high temperature tolerant plant growth promoting rhizobacteria (PGPR) in alleviation of heat stress adverse effect in eggplant.

Methods: PGPR was assessed for PGP potentialities and investigated for1‐aminocyclopropane‐1‐carboxylate (ACC deaminase), and exopolysaccharide under normal and high temperature environment. The impact of PGPR application on plant physiology and biochemistry was investigated under normal and high temperature environments and plant growth regulators were evaluated using High Performance Liquid Chromatography (HPLC).

Results: A significant impediment for morphological and physiological parameters of eggplant cultivars was observed during heat stress in absence of PGPR inoculation. The results showed that ACC‐deaminase produced by PGPR boosted up significantly the extracellular polymeric substances (EPS) accumulation, conversion of ACC into α‐ketobutyrate and ammonia, and reduce consequently the impact of high temperature on eggplant development.

Conclusion: PGPR inoculation showed an alternative strategy to improve eggplant growth and development under high temperature condition in order to preserve agriculture sustainability and healthy crops. 

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Iron-Based Nanoparticles Synthesis, Characterization, and Antimicrobial Effectiveness
Hisham Faiz Jaha, Yasir Anwar, Saleh M. Al-Maaqar, Tahseen Kamal, Sher Bahadar Khan, pages 525-532
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 Background: Antimicrobial resistance (AMR) has emerged as a significant and pressing public health concern, posing serious challenges to effectively preventing and treating persistent diseases. Developing new antibiotics with different mechanisms of action is crucial to effectively address challenges in treating infections. A lot of work has already been done on mono-metallic nanoparticles to address the issue.

Methods: This study aimed to synthesize multi-metallic iron, silver, and chitosan-embedded nanoparticles using a green approach. Iron, silver, chitosan nanoparticles, and a composite of iron–silver–chitosan was also synthesized. The synthesized nanoparticles and composites were characterized through X-ray diffraction (XRD), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), and Fourier Transform Infrared Spectroscopy (FTIR) to evaluate their structural parameters. Their antimicrobial efficacy was investigated through MIC (minimum inhibitory concentration), MBC (minimum bactericidal concentration), and well-disk diffusion assays against Pseudomonas aeruginosa, Acinetobacter baumanniiStaphylococcus aureus,  Staphylococcus epidermidis and Candida albicans.

Results: The size of the Cu-NPs, Cu-Ag NPs, and Cu-Ag-CS NPs were found to be in the range of 32-40 nm size with a spherical shape. The nanocomposites' MIC and MBC were calculated to be 125 μg/mL and 500 μg/mL, respectively. The nanocomposites exhibited a range of clear inhibition zones, with a minimum diameter of 12 ± 0.5 mm and a maximum diameter of 22 ± 0.5 mm. 

Conclusion: The iron–silver–chitosan nanocomposite has been shown to have significant antimicrobial effects in the laboratory environment compared to other nanoparticles hence can be applied as potential biomedical/biological candidates in future. 

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Developing a Nano Platform for Bovine Brucellosis Diagnostic Product
Gulmira Tulepova, Edil Makhashov, Loukia Ekateriniadou, Nurzhan Nurkhojayev, Sagypash Sadiev, pages 533-538
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 Background: Brucellosis is one of the most common zoonoses caused by Brucella species. Brucella is an intracellular pathogen that causes abortion in domestic animals and brucellosis in humans. The disease has no specific pathognomonic signs; hence a prompt and accurate diagnosis of the disease plays a very critical role in preventing brucella transmission and treating the disease.

Methods: Research was carried out at the Department of Veterinary Science of Kazakh National Agrarian Research University and in the Molecular Biology laboratory of the Institute for Plant Biology and Biotechnology under the Committee of Science of the Ministry of Education and Science of the Republic of Kazakhstan. Studies were carried out on a killed culture of the vaccine strain Brucella abortus RB19, genomic DNA of B. abortus RB19, Grapevine virus A, plasmid vectors for DNA cloning, a fluorescent nano marker, and Brucella membrane proteins.

Results: The research demonstrated that brucellosis remained one of the most common zoonotic diseases worldwide. This disease not only resulted in significant economic losses in agriculture but also posed a serious threat to public health.

Conclusions: The study successfully established prokaryotic expression vectors for efficient protein production, validated through sequencing and Western blotting, and proposed a promising nano-platform for plant-based synthesis of brucellosis proteins, with potential implications for diagnostic kit development and vaccine research in agriculture. 

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