Volume 11, Issue 3

Advancements in Life Sciences, volume 11, issue 3 – August 2024cover
ISSN 2310-5380

IN THIS ISSUE

This 3rd issue of the 11th volume has featured 26 multidisciplinary articles including 4 review articles and 22 research papers of authors from 13 countries covering aspects from different stems of life science. Enjoy reading!

 

 

 Review Articles


Prevention of ear‐looped face mask‐induced pressure injury on ears: A Technical report with review of literature

Fareedi Mukram Ali, Abdullah Saeed Wasli, Abbas Hasan Hobani, Samira Hussain Al Faraj, Essa Hamoud Mashiakhy, Ali Mohammed Alkhayrat, Reem Abdullah Khawaji, Reem Abdullah Khawaji, pages 539-543
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 In modern times, protective masks with flexible loops are commonly used to prevent the spread of microorganisms during surgical procedure the constant contact of the stretching bands caused by wearing a mask for several hours’ daily causes pressure on this cartilage, leading to painful and erythematous lesions on the skin behind the auricles. Patients and the public now wear masks for extended periods of time. Over time, wearing this mask frequently can cause damage to the ear. Consequently, people regularly switch out their masks to alleviate pressure, increasing their chances of getting infected. Doctors and other healthcare professionals should be aware that continued use may cause damage to the ears and auricles. This article addresses a novel method to alleviate stress on the ear loops of surgical masks and examines the typical ear injuries or pressure injuries on the ear mentioned in literature regarding prolonged mask usage and ways to prevent them. 

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Microbial forensics: are we going on right path for management of crops biosecurity in Pakistan?
Ishrat Aziz, Saadat Ali, Zia-ur-Rehman, Sitwat Aman, Ahmed Ali Shahid, pages 544-548
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 In any country biological terrorism outbreaks on crops could reduce the quality and quantity of agricultural products, which in turn decreases the consumer’s confidence, nutrition loss, resulting in worse impacts on the nation’s economy, international souks and is subsequently harmful for the environment. Awareness for a crop bioterror incident begins with the start of the threat symptoms, which must lead to the progress of a plan to thwart. However, most of Pakistani scientists and crop producers have not focused on the option of deliberative plant pathogen introduction but instead they use their expertise for the prevention of natural introductions and the improvement of cost-effective strategies for disease management.  It is also an interesting fact that under the mask of genetically modified feeds, high yield varieties and hybrids; we are also importing bio terrorism. Are we going on the right path?  So, it is time that we should review the global microbial knowledge and should apply it according to our needs. In ongoing topics of present article, we will review the microbial forensics status globally in comparison with Pakistan. We will also suggest how to fill these gaps. 

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Commands of Synthetic Biology to Modernize and Re-design the Biological Systems
Hina Ilyas, Aimen Afzal, Zaheer Abbas, Sabahat Noor, Irfan Ullah, Raja Sheraz Rafique, Zeeshan Abbas, Muhammad Umer Bin Muhammad Ishaq, pages 549-557

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 The scope of synthetic biology continues to expand and has encompassed a huge number of biological features. Its scope starts from scratch, enabling the de novo synthesis of biological systems. It has re-designed the biological systems and empowered the production of synthetic genes, RNA, DNA and proteins by undertaking the control of pathways involved in genetic regulation. It has increased the production of nano-scale RNA architectures and synthetic biological circuits which either have therapeutic or other productive uses. Furthermore, advancements in synthetic biology have enabled the generation of diversity through methods such as epPCR and site-directed mutagenesis, allowing for the creation of complex genetic variations. Additionally, synthetic biology intersects with computer engineering to design functional biological devices and circuits, utilizing computational analysis to guide the design process. Moreover, ethical and regulatory considerations are paramount in synthetic biology, with careful examination required to address dual-use concerns, environmental impacts, and issues of social justice and equitable access to benefits. As synthetic biology continues to advance, it presents opportunities to address pressing challenges in fields ranging from medicine and agriculture to environmental conservation and beyond. Thus, the fusion of synthetic biology with other scientific disciplines holds promise for transformative innovation and societal benefit. The present discussion enlightened the core of generating complex biological systems and has given a brief overview on the fusion of synthetic biology with other fields of science. 

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Effect of Supportive Care on Quality of Life for People Living with Human Immunodeficiency Virus: A Systematic Review
Muh. Yusuf Tahir, Muh. Zukri Malik, I Kade Wijaya, Kusrini S Kadar, Risna Halim Mubin, Firdaus Hamid, pages 558-564

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 The scope of synthetic biology continues to expand and has encompassed a huge number of biological features. Its scope starts from scratch, enabling the de novo synthesis of biological systems. It has re-designed the biological systems and empowered the production of synthetic genes, RNA, DNA and proteins by undertaking the control of pathways involved in genetic regulation. It has increased the production of nano-scale RNA architectures and synthetic biological circuits which either have therapeutic or other productive uses. Furthermore, advancements in synthetic biology have enabled the generation of diversity through methods such as epPCR and site-directed mutagenesis, allowing for the creation of complex genetic variations. Additionally, synthetic biology intersects with computer engineering to design functional biological devices and circuits, utilizing computational analysis to guide the design process. Moreover, ethical and regulatory considerations are paramount in synthetic biology, with careful examination required to address dual-use concerns, environmental impacts, and issues of social justice and equitable access to benefits. As synthetic biology continues to advance, it presents opportunities to address pressing challenges in fields ranging from medicine and agriculture to environmental conservation and beyond. Thus, the fusion of synthetic biology with other scientific disciplines holds promise for transformative innovation and societal benefit. The present discussion enlightened the core of generating complex biological systems and has given a brief overview on the fusion of synthetic biology with other fields of science. 

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Full Length Research Articles


Effect of Supportive Care on Quality of Life for People Living with Human Immunodeficiency Virus: A Systematic Review
Muh. Yusuf Tahir, Muh. Zukri Malik, I Kade Wijaya, Kusrini S Kadar, Risna Halim Mubin, Firdaus Hamid, pages 565-571

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 Background: There are many weed species that affect yield and quality in date palm orchards. Cutandia memphitica (Spreng.) Benth. is one of the weeds that is common in date palm orchards in Iraq. However, there is not enough information about C. memphitica in the literature. In order to provide information about the effect of key environmental factors (temperature, light, pH, and salt) on the seed biology of this weed species, this study was conducted.

Methods: Within 2020, seeds were collected from agricultural regions in southern Iraq, and trials were done at Erciyes University Faculty of Agriculture labs in Kayseri Türkiye. All experiments were carried out with 30 seeds in the sterile 90 mm (90×15) petri dishes containing two layers of sterile filter paper.

Results: The results of this investigation revealed that the seeds of this species germinated by 72.49% in three days when incubated at 25°C, and by 96.66% in 14 days when incubated at 15°C. The results also indicated that even during the longest incubation time examined, which was 28 days, the seeds of this species never germinated at 30 degrees or above. They can germinate during this period if kept at 5°C. The study also revealed that 90% of the seeds of this species germinated in complete darkness, whereas only 11.66% germinated in light. However, light has a significant impact on studied growth parameters (plant height and seedling fresh and dry weight). The results also demonstrate that the pH levels examined, which varied from 4 to 10, had no effect on the seed germination and growth parameters of seedling.

Conclusion: The results showed that the seeds of this species germinated at the highest salinity levels (250 mM NaCl) by 10.83%. Meanwhile, the best germination was recorded at 25 mM salinity level of 89.16%, but without significant differences with salinity level at 50 mM and control. Also, the growth indicators were reduced significantly at the salinity level of 250 mM compared to the control. 

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Metal Sulfates and Microbial Food Treatment Alleviate the Oxidative Damage Caused by PEG-Induced Osmotic Stress in Cotton Plants
Semra Almas, Tijen Demiral Sert, pages 572-579

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 Background: Drought stress restricts cotton production, which causes a considerable loss of cotton yields worldwide. Plant activators improve crop production and ameliorate abiotic stresses. Thus, the effects of plant activators on the genotype of cotton (Gossypium hirsutum L.) Candia, which is resistant to drought, were investigated.

Methods: Plants were treated with the plant activators, and then osmotic stress was initiated by 30% PEG 6000 treatment for 13 days. Antioxidant enzyme activities such as superoxide dismutase (SOD), ascorbate peroxidase (APX), and peroxidase (POX) were measured along with growth, relative water content (RWC), osmotic potential, photosynthetic pigments, ion leakage, proline accumulation, malondialdehyde (MDA) content, and reactive oxygen species (ROS).

Results: Osmotic stress reduced shoot fresh weight (FW) and osmotic potential but increased root length of Candia. Activator treatment alleviated these effects. Photosynthetic pigment contents and cell membrane permeability were not affected by osmotic stress and activator treatments; proline accumulation and lipid peroxidation levels were enhanced by osmotic stress, but activators alleviated these increments. Activity bands for the isozymes Mn-SOD1, SOD2, and SOD3, as well as the isozyme Fe-SOD, were found in Candia. Osmotic stress elevated APX activity in Candia leaves, while activator treatment decreased it. POX activity was not affected by osmotic stress, but activator treatment decreased POX activity. Osmotic stress-induced accumulation of ROS was lessened by activator treatment.

Conclusion: Plant activator treatment reduced the amount of oxidative damage (ROS buildup and lipid peroxidation) caused by drought stress in the Candia genotype. 

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Clinical study of common bacterial, fungal and parasitic skin diseases in cats
Hassanin H. N. Alautaish, Hussein Ali Naji, Zainab Abudal Hussein Saud, Baneen Ghanim Ghalib, pages 580-584

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 Background: The skin is a big, metabolically active organ that has a significant need for other nutrients and protein in its body. The study pointed high significant infection with fungal, scabies, alopecia, bacterial and wounds in the white color cats more than the others because it is represented the better color choice for the families then the orange and grey colors while the black color is the less in number of domestic cats. The aim of present study was investigated of some skin disease in cats in Basrah, Iraq.

Methods: The present study includes two hindered animals of both sexes with different ages, colors, weights and sizes were studied clinically in Basrah veterinary hospital and the private veterinary clinics in Basrah. The study extended from October 2022 to March 2023 during the winter season. Clinical signs were reported carefully, and the initial diagnosis of skin diseases was confirmed by apparent lesions on the skin, the skin samples of the cases were collected for laboratory diagnosis in veterinary medicine college ̸ Basrah University and Basrah veterinary hospital laboratories to confirmed diagnosis.

Results: Clinical signs were reported carefully and the initial diagnosis of skin diseases. The two hindered sample cases were divided according to cases in Basrah veterinary hospital and Basrah private clinic: 60 Cases fungal infection, 40 cases scabies, 10 case alopecia, 50 cases of bacterial infection and 40 cases of wound. 

Conclusion: The present study shows the fungal diseases were common causes for skin diseases in Cats follow by Bacterial, Parasitic and Alopecia respectively. 

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Construction of subunit-based DNA vaccines against tuberculosis using five different Mycobacterium specific genes
Muhammad Mohsin Zaman, Mirza Imran Shahzad, Aeman Jilani, Areeba Yousaf, Ansah Bashir, Saher Riaz, Gildardo Rivera, pages 585-592

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 Background: Tuberculosis remains the major infectious and contagious disease with respect to morbidity and mortality around the globe. The main etiological agent of TB is Mycobacterium tuberculosis (M. tb). The cure and/or control of this disease is getting difficult day by day especially due to emergence of drug resistant strains, HIV co-infection and unavailability of good vaccines. BCG is the only available and permitted vaccine against TB, which has variable efficacy only limited to childhood. Therefore, new, well defined, antigen specific and state of the art Molecular Biology based DNA vaccines are required.

Methods: The current study is designed to develop subunit-based DNA vaccines against tuberculosis by using five different Mycobacterium specific genes, namely Rv1908c/KatG, Rv0350/DnaK, Rv0440/GroEL2, Rv0934/PstS and Rv3418c/GroES. All the selected genes were amplified through PCR and cloned into mammalian expression pVAX1 vector.

Results: All the transformed constructs were confirmed through restriction digestion, colony PCR and sequence analysis. These constructs will be used for various in Vivo immunization studies and also for the challenge studies against TB in future experiments.

Conclusion: DNA vaccines, alone or in combination with BCG, have enough potential to be a good therapeutic tool for TB and reduce the treatment time in future. 

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Soluble endoglin as a diagnosis glycoprotein in preeclampsia
Shaima Sh. Al-Kilan, Ekhlas Abdallah Hassan, pages 593-599

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 Background: Angiogenic factor imbalance, such as that caused by soluble endoglin (sEng), is a feature of preeclampsia. However, the connection between sEng and clinical and laboratory indicators, and the severity of preeclampsia is not entirely understood.

Methods:  Ninety subjects were incorporated in this study, 30 were healthy pregnant with mean age (35.6) years. (32) mild  Preeclampsia case with mean age (32.65) years preeclampsia, and (30) sever PE with mean age (32.65) wherever all women were more than 24 weeks of pregnancy. Mean blood pressure and level of proteinuria were used as indicators of the severity of the disease. ELISA was used to measure the levels of sEng in the serum. Serum lipid profile was measured by enzymatic methods. The qualitative dip-stick technique (CYBOWTM DFI Co Ltd, Republic of Korea) was used to assess the amount of urine protein.

Results : A substantial difference between the PE group's serum sEng concentration and that of the healthy subjects (p 0.001) could be seen. Serum sEng concentrations of patients with moderate PE and those with severe PE differed significantly. Additionally, there were strong positive relationships between the serum sEng concentration and the SBP and proteinuria. serum sEng levels, biochemical indicators, and other factors, however, did not significantly correlate. The diagnostic accuracy (86.2) in distinguishing mild PE from the healthy patient group was good thanks to the excellent area under the curve (AUC = 0.827, p < 0.0001).  

Conclusion: Levels of  sEng  in sera of PE patients were elevated. sEng was significantly correlated with PE patients. 

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Clinical, hematological and some biochemical alterations of Rotavirus group A in newborn buffalo calves
Asma Salim Tarik, Muhsen RK, pages 600-606

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 Background: Infectious diarrhea of newborns calves is one of the biggest health problems in beef and dairy industries in Basrah, Iraq, Group A rotavirus considers major cause of gastroenteritis in newborn calve  of buffalo in the world, leading to serious economic losses specifically in developing countries. The infection has short incubation period, non-viremic, and profuse diarrhea.

Methods: Buffalo calves with diarrhea were carefully examined and a sample of their excrement was collected for this particular study. The calves ranged in age from one to thirty days and were equally split between males and females. We found out whether the calves were girls or boys. The calves' ages were considered throughout the sorting process; they originated from different regions of Iraq's Basrah governorate. Moreover, they were further separated into four separate age groups. Blood samples were collected from calves infected with rotavirus so that the objectives of this research could be fulfilled. Biochemical and hematological studies were the intended purposes of the blood sample collection. Finding out how these samples fared in comparison to the control samples in the group was the next stage. Laboratory diagnostics used real-time polymerase chain reaction (PCR), while field diagnostics used a rotavirus antigen fast test kit. Both techniques were used in tandem.

Results: With the use of a field-based rotavirus antigen fast detection kit and real-time polymerase chain reaction, 19 out of 30 samples tested positive for rotavirus antigen. Nineteen out of one hundred samples came back positive. Along with it, the highest infection rates were recorded in January (30.7%) and among children aged five to fourteen (27.5%). The afflicted males and females did not vary significantly from one another statistically. The clinical signs of infected calves were also documented. Symptoms included anorexia with or without a loss of suckling ability, lose or somewhat solid stools, mucous, and a milky or light-yellow color. Some of the other symptoms included signs of dehydration, although to varying degrees. Nothing stood out while comparing the patients' vital indicators. In the infected group, sodium levels dropped significantly, and monocyte and lymphocyte production skyrocketed. If we look at potassium levels, we see that there was no statistically significant difference between the infected and control groups. Infected calves had substantially greater levels of red blood cell count, panel C viral load, hemoglobin B, and total lipid content (TLC) when compared with healthy calves.

Conclusion: The study record and suggests the role of rotavirus group A as a serious cause of diarrhea in newborns buffalo calves in Basrah governorate, Iraq. 

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Extraction and Partial Purification of Glucoamylase from Rhizopus oryzae by Solid State Fermentation using Agro-Industrial Residues
Qays Majeed Issa, pages 607-612

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 Background: Filamentous fungi are used to produce many industrial enzymes because these fungi have a great ability to produce such enzymes, especially when using solid substrates that are usually low in cost. The widespread use of this enzyme has made it very important to work on improving its production to achieve the maximum possible benefit.

Methods: Five fungal isolates were obtained from soil and decaying fruits and vegetables. All isolates were identified as Rhizopus spp. and cultured on Potato Dextrose Agar (PDA) plates escorted by starch 1%. The appearance of clearance zones around the fungal colonies represents the ability of fungi to produce Glucoamylase. Different solid substrates, nitrogen sources, and temperatures were used to improve enzyme production.

Results: Wheat bran gave the highest enzyme production with specific activity 6.25 U/mg, and yeast extract was the potent inducer for enzyme production with specific activity 9.28 U/mg. The optimal temperature for enzyme production was 30°C with specific activity10.27U/mg. Maximum specific activity 18.40U/mg was recorded at 55% saturation with ammonium sulfate precipitation. The result showed increased specific activity 24.84U/mg with dialysis. Partial purification of the enzyme revealed that pH5 was optimum for enzyme-specific activity 9.2U/ml; higher enzyme-specific activity was found with MnSO4 (11.83U/ml) as the best enzyme inducer.

Conclusion: This investigation attempted to examine R. oryzae as an effective maker of GA. Innovative work into the nearby creation of GA for industrial use utilizing local resources has demonstrated financial effectiveness. It requires improvement into consistency as needed by worldwide associations dealing with industrial enzymes.  

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Exploring Gene Polymorphisms Associated with Otitis Media through Variable Number Tandem Repeats (VNTR) Region
Safa Amer Khalil, Kawther M. A. Hasan, pages 613-618

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 Background: Otitis is an inflammation and infection of the inner lining of the middle ear. The problem known as suppurative otitis media if there are droplets and holes in the tympanic membrane due to this inflammation and infection of the mucous membrane. The present study focused on specific genes suspected to be associated with otitis media in 60 subjects clinically diagnosed with OM The study focuses on two genes, MUC5B and IL-1RN, whose genes mutations by variable number of tandem repeat (VNTR) regions

Methods: Variable numbers of tandem repeats (VNTR) regions were used to genotype 60 patients with otitis media and 30 healthy individuals for genotypic polymorphisms for the MUC5B and IL-1RN genes by number of tandem repeats (VNTR) regions on a variable basis Blood samples were collected from participants, and three milliliters were placed directly into EDTA tubes. Subsequently, PCR products were detected by agarose gel electrophoresis and visualized by ethidium bromide staining. The size of amplified DNA fragments was determined by comparison with a 100-bp DNA ladder molecule size marker

Results: About the same, the paper represents the association of gene polymorphisms with otitis media by genotyping a total of 60 patients as OM, and, on the other hand, 30 individuals were noted to be healthy. This was to determine genotypic polymorphisms for the two genes, that is, MUC5B and IL1RN, within the VNTR region. The results showed that the carriers of allele 2 in the genotypic position had a risk of nearly twofold getting otitis media.

Conclusion: Polymorphism of the MUC5B and IL-1RN genes can be responsible for the severity of the inflammatory process in the middle ear, complicated by otitis media or hearing loss. 

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Separation and partial purification of lecithin: cholesterol acyltransferase from serum of obese women with a study of the effect of oily and nano-extract of Castanea fruit in activating the enzyme
Intisar Ghanim Taha, Eman Salem Mahmoud, Shemaa Abass Ayob, pages 619-623

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 Background: The Chestnut plant is one of the important plants for treating many diseases. Therefore, the oil extract of the Chestnut plant was taken to know its effect on activating an enzyme of lecithin: cholesterol acyltransferase that decreases in obese patients, by isolating it from the blood and using nanotechnology methods.

Methods: The research included the isolation lecithin: acyltransferase cholesterol (LCAT) from the blood of 5 obese women, whose BMI was between 25-30 kg/m2, ages ranged from 40-45 years, and they were healthy without any diseases. The oil extract of Chestnut fruit was isolated. Fatty acids present in the oil extract were identified using capillary gas chromatography technique CGC, as the extract contained a percentage of unsaturated fatty acids and a lower percentage of saturated fatty acids. The enzyme was precipitated with ammonium sulfate, dialysis technique, and the gel filtration chromatography technique was used in purification. Finally, effect of the oil extract of the Chestnut fruit was studied at a concentration of (0-100) mg/ml and the nanoparticles-oil extract also on the activity of the LCAT (0-20) mg/ml. The nanoparticles of silver nitrate were prepared at a concentration of 1 mM with the oil extract of the Chestnut fruit and heated at a temperature of 60°C.

Result: The highest peak was obtained from the gel filtration chromatography, which was highly effective. The number of purification times reached 91.72, and the recovery rate was 283.33%, and the effectiveness was qualitative 457.69 U/mg. Oil nanoparticles extraction was confirmed when the solution was transformed by changing its colors from colorless, yellow, orange, and finally coffee in color. The results showed an increase in the activity of the LCAT when treated with the two extracts, also the increase in the activity had a higher effect on the activity of the enzyme.

Conclusion: We concluded during the study that the extract of the fruit of the oily nanoparticles was more effective in increasing the activity of an enzyme lecithin: cholesterol acyltransferase in healthy women with obesity.  

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Comparative Assessment of Gutta-Percha Removal Techniques for Endodontic Retreatment: Efficiency and Clinical Implications
Ashwaq Asiri, pages 624-628

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 Background: Endodontic retreatment involves the removal of gutta-percha and sealer from root canals to address treatment failures and persistent infections. This study aimed to systematically evaluate and compare the effectiveness of different gutta-percha removal techniques.

Methods: Sixty extracted human teeth with single canals were selected. The canals were initially treated, obturated with gutta-percha, and divided into six groups (n=10 each). Each group represented a distinct removal technique: rotary instrument-driven, Gates-Glidden drill-assisted, solvent-based dissolution, ultrasonic removal, heat-based softening, and a control group. Gutta-percha mass and canal wall cleanliness were assessed post-removal. Statistical analyses were performed using ANOVA and post hoc Tukey's HSD tests.

Results: The solvent-based dissolution technique exhibited the most efficient removal, achieving the lowest mean remaining gutta-percha mass (0.89 ± 0.15 mg) and the highest canal wall cleanliness (95% ± 3%). The ultrasonic technique also showed effective removal capabilities with a mean remaining mass of 1.56 ± 0.22 mg and a cleanliness of 89% ± 4%. The rotary technique resulted in a mean mass of 2.47 ± 0.30 mg and cleanliness of 84% ± 5%. Gates-Glidden drill-assisted and heat-based techniques showed moderate performance with remaining masses of 3.18 ± 0.25 mg and 2.10 ± 0.20 mg, respectively, and cleanliness percentages of 78% ± 6% and 81% ± 5%. The control group had the highest remaining mass (9.88 ± 0.42 mg) and the lowest cleanliness (10% ± 2%), confirming the efficacy of the removal techniques.

Conclusion: The solvent-based dissolution technique demonstrated superior efficacy in gutta-percha removal. Ultrasonic and rotary techniques were also effective choices. Clinicians should consider these findings when selecting gutta-percha removal methods for endodontic retreatment procedures. 

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Chronic sleep deprivation, noise and high fat diet markedly induce C-reactive protein level in mice model
Sama Attiyah, pages 629-633

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 Background: C-reactive protein (CRP) is a circulating sign of systemic chronic inflammation that your liver produces if your body is inflamed. Stressful events over a long period of time are linked to inflammatory disorders. Stress pathways are triggered by noise and sleep deprivation, as well as unhealthy habits like fatty meals. Several studies have connected higher CRP levels with plasma levels. The current study aims to explore the potential association’s relationship between chronic sleep deprivation, noise, high fat diet, and CRP disorders.

Methods: Three studies were carried out in our study to evaluate C-reactive protein levels in male Swiss albino mice groups after exposure to three different physical stressors for four weeks. The first study mice group were exposed to chronic sleep deprivation; second group was exposed to electrostatic speaker noise stressors; and the third group was fed a high fat diet. Quantitative estimation of C – reactive protein level in plasma using an ELISA kit.

Results: Significant changes in inflammatory CRP levels in the examined plasma were detected in the sleep deprivation group. CRP levels were found to be steadily increasing after two weeks, reaching a significant peak in the fourth week (p < 0.001). Moreover, in the fourth week, there were substantial increases in CRP plasma level with independent evidence, in the experimental noise group (p < 0.001). As well, the biomarker of inflammation level increased across the dietary high density fat diet, reaching the highest level at the end of the study (p < 0.05).

Conclusion: Our findings contribute to the body of evidence indicating a link between external stress and subclinical inflammatory markers. Furthermore, CRP concentrations are triggered by chronic sleep deprivation and noise exposure, and their concentrations rise with prolonged dietary fatty acid consumption. 

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Exploring Listeria monocytogenes in Ewe Milk: ssrA Gene-based Real-time PCR Identification, Phylogenetic Analysis, and Antibacterial Assessment of Magnesium Oxide Nanoparticles Synthesized with Myrtus communis Leaf Extract
Jenan Nadhim Sadeq, Balsam Miri Mizher, Alyaa Abdulhussein Alsaedi, Ola Hakim Khudhair, pages 634-640
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 Background: Listeria monocytogenes, a zoonotic pathogen affecting humans and animals, exhibits a global distribution, including Iraq. This study focused on the rapid identification and phylogenetic analysis of L. monocytogenes in freshly collected ewe milk samples from various farms in Al-Qadisiyah province, Iraq.

Methods: The study was conducted with care and precision, involving 150 milk samples. These samples were subjected to traditional bacterial isolation and identification using the enrichment culture method and biochemical tests, with the PCR technique confirming the results. The antibacterial activity of MgONPs was then assessed using the disc diffusion method, ensuring a comprehensive and reliable approach to the study.

Result: The results show that 150 ewe milk samples underwent real-time PCR (RT-PCR) targeting the ssrA gene, followed by partial 16S rRNA gene sequencing (PSGS) of purified conventional PCR products. Furthermore, the study entails the biosynthesis of magnesium oxide nanoparticles using Myrtus communis  leaf extract, followed by a comprehensive characterization utilizing UV-spectra, FTIR, SEM, and TEM techniques. The Agar well diffusion method assessed the antibacterial efficacy of these Biosynthesized nanoparticles against L. monocytogenes. The RT-PCR results revealed the presence of L. monocytogenes in 36 out of 150 samples (24%). Subsequent PCR analysis confirmed the presence of the pathogen in 30 out of these 36 positive samples (83.33%). Sequencing of two purified PCR products demonstrated 100% nucleotide identity with global isolates from Iraq and Turkey. Furthermore, the study demonstrated that L. monocytogenes exhibited substantial sensitivity (24.66 ± 0.3) to the biosynthesized magnesium oxide nanoparticles. These findings underscore the speed and precision of the RT-PCR method for detecting L. monocytogenes in fresh ewe milk samples.

Conclusion: This comprehensive investigation enhances our understanding of L. monocytogenes prevalence in ewe milk and highlights the potential of Myrtus communis -derived nanoparticles for combating this pathogen. 

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Evaluation of Micro-RNA Expression Profiling Level as Biomarkers for Diagnosis and Gene Sequencing in Patients Suffering from Breast Cancer
Rasha H. Al-Rikabi, Nagui H. Fares, Mahmmad A. AL Faham, Abdel Hady A. Abdel Wahab, pages 641-647
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 Background: Human bloodstream microRNAs (miRNAs) have emerged as a promising predictive and diagnostic biomarker for a range of cancers, including breast cancer. Our objective was to look into new miRNA biomarkers for diagnosis in the serum of patients with breast cancer and track the expression levels at different stages using miRNA profiling.

Methods: 53 breast cancer patients and 10 healthy controls had blood samples tested for three miRNAs. miRNAs were extracted from blood and evaluated using real-time quantitative polymerase chain reaction. After extracting genomic DNA, miRNA primer-produced PCR products were sequenced to discover point mutations that may contribute to the illness.

Results: After examining miR195, miR200b, and miR331, breast cancer patients had a significantly lower miR195 level than healthy persons. In addition, miR200b expression levels were significantly lower in breast cancer patients than in healthy individuals. In advanced stages, miR331 expression was substantially higher than in healthy people.

Conclusion: The findings of our study demonstrated a significant association between the expression of miRNAs and the prognosis of breast cancer. Additional research is necessary to study better the correlation between these circulating miRNAs and the stages of breast cancer. 

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Evaluation of BACTEC™ FX and BacT/Alert™ Automated Blood Culture Systems for Detection of Clinically Relevant Bacterial and Yeast Species
Muhammad Absar, Ibraheem Alshahrani, Fahd Nashmy, Mosa Salami, Ali Hadadi, Aous AlMarzooq, Alhussain AlAsiri, Abdulaziz AlRashidi, Mohammed AlFaifi, pages 648-654
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 Background: Bloodstream infections (BSI) are a significant health concern, necessitating effective diagnostic tools. This study evaluates the performance of the BD Bactec FX system in comparison to the established BacT/Alert system, examining mean time till detection (TTD) across various blood culture bottles.

Methods: The study involves 148 blood culture bottles, inoculated with representative bacterial ATCC strains (n= 15) and 04 yeast isolates. Parallel testing is conducted by seeding 10–30 colony-forming units (CFU) in duplicate in both BD Bactec FX and BacT/Alert systems. TTD is assessed across aerobic, anaerobic, and pediatric bottles for diverse microbial species.

Results: All 148 bottles tested in parallel show positive signals in both systems. BD Bactec FX demonstrates significantly shorter TTD for adult-seeded cultures in aerobic and anaerobic bottles compared to BacT/Alert Similarly, pediatric bottles with BD Bactec FX exhibit a shorter TTD compared to BacT/Alert. The statistical significance of TTD, 95% confidence intervals (CI), and p-values is evident for Bactec FX aerobic, anaerobic, and pediatric bottles across tested organisms. Notable examples of faster TTD include; Bacteroides ovatus (16.6h by Bactec FX vs. 70.5h by Bact/Alert 3D), Stenotrophomonas maltophilia (33.7h vs. 72.5h), and Streptococcus pyogenes (8.1h vs. 11.9h).

Conclusions: In conclusion, the study’s findings demonstrate that BD Bactec FX surpasses BacT/Alert in prompt microbial detection, showcasing potential for early identification of bacteremia and fungemia. Faster TTD implies the potential to initiate timely antimicrobial treatment, thereby reducing patient morbidity and mortality. However, exceptions in certain microbial species highlight the need for comprehensive clinical validation to establish the broad applicability of these findings. 

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Serum 25-Hydroxy Vitamin D and Uric Acid Levels as Predictors of Disease Severity and Functional Impairment in Women with Knee Osteoarthritis
Amal F. Gharib, Ahad A. Alsaiari, Renad A. Alshamrani, Norah M. Al-Qahtani, Abdulraheem Almalki, Ohud Alsalmi, Mazen Almehmadi, Norah Al harthi, Asmaa F. Hassan, Alaa Hamid Alsulimani, Enas I. Abdelhady, Ola Nafea, pages 655-662
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 Background: The most frequent type of arthritis is osteoarthritis; it typically develops slowly and is more frequently observed in older adults. Osteoarthritis mainly affects weight-bearing joints but other can also affected joints.  The purpose of the study was to explore the associations between serum of 25-hydroxy vitamin D (25(OH)D) and serum uric acid (SUA) with serum lipid profiles and inflammatory cytokines in women with KOA and to scrutinize the prognostic potential of serum 25(OH)D and SUA.

Methods: The study involved 50 elderly obese females with KOA and the control group of 50 healthy women. Assessment of KOA severity by Kellgren-Lawrence score was performed and functional disability was scored. Blood samples were analysed to assess 25(OH)D and inflammatory cytokines, SUA, lipid profile, and calcium.

Results: We found significant differences in most of the laboratory findings between KOA patients and controls. Decreased serum 25(OH)D and increased SUA were linked with more severe KOA. Serum 25(OH)D and SUA can differentiate between mild and moderate-to-severe KOA. Both were correlated with functional impairment.

Conclusions: We concluded robust associations between vitamin D deficiencies, high SUA levels with increased serum lipids and inflammatory cytokines along with increased functional impairment and disease severity in women with KOA. Serum 25(OH)D and SUA can serve as simple reliable prognostic tools in the prediction of KOA severity. 

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First Report on Clinical Feasibility of Dried Blood Spot Technique for Hemoglobin Estimation in Cholistani Cattle
Maryam Chaudhary, Umer Farooq, Musadiq Idris, Mushtaq Hussain Lashari, Saira Qasim, Muhammad Abrar Afzal, Musarrat Abbas Khan, Asad Ali, pages 663-668
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 Background: The dried blood spot (DBS) technique using filter papers has revolutionized the conventional blood sampling techniques through ease of blood collection, storage and transport. Various analytes (such as hormones, antigens, antibodies and hematochemical attributes) are being estimated through DBS globally. However, this technique has not yet been implied in Pakistan. This research work is the first of its kind regarding hemoglobin (Hb) estimation in Cholistani cattle (n=63) blood through DBS technique using filter paper.

Methods: Three methods of Hb estimation were implied in the present study viz. through veterinary hematology analyzer (HbA), and two indirect cyanmethemoglobin methods (HbIC and HbICX) using measured (20µL) and unmeasured blood drops on the filter paper, respectively.

Results: Results revealed that HbA and HbIC were non-significantly (P≥0.05) different from each other, being different from HbICX (P≤0.05). The HbICX gave overestimated values of Hb as compared to HbA and HbIC. The sensitivity, specificity, positive predictive value, and negative predictive value for HbIC were higher being 86.1%, 88.3%, 88.0%, and 86.0%, respectively as compared to the lower values of 45.0%, 12.0%, 12.0% and 45.0%, for HbICX. Bland and Altman test revealed a better level of agreement between HbA and HbIC. Around the mean difference line, there was no proportional bias in data distribution (Mean= -0.16, 95% CI= 0.34 to -0.67). Similarly, measures attained through Cronbach alpha and intraclass correlation coefficient between HbA and HbIC were higher being 0.703 and 0.825 values for single and average, respectively, as compared to the values of 0.200 and 0.333 between HbA and HbICX.

Conclusion: It is concluded that the indirect cyanmethemoglobin method for Hb estimation is reliable and accurate for cattle blood, if a measured quantity of blood drop is taken on a filter paper. We recommend this DBS technique for Hb estimation in cattle blood for resource-poor settings and for livestock herds being reared distantly from the laboratories. For future, it is recommended that DBS technique with various other modifications and for other hematochemical biomarkers may be validated for livestock blood. 

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Antidiabetic Activity and Pancreatic Histopathology Analysis of Averrhoa bilimbi L. Leaves Extract on Alloxan-Induced Diabetes on Wistar Mice
Tridiganita Intan Solikhah, Ragil Kusnandar Miftakhurrozaq, Aswin Rafif Khairullah, pages 669-673
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 Background: Matrix metalloproteinase-2 (MMP2) plays a role in breaking down the components of the extracellular matrix, which allows cancer cells to advance and invade. Therefore, the inhibition of MMP2 shows potential as a promising strategy for treating cancer.

Methods: This study employed computational screening to identify MMP2 inhibitors from a collection of 2,405 natural compounds. GLXC-26716, the co-crystal ligand of MMP2, served as the positive control. Virtual screening was performed using PyRx 8.0 software to find molecules that might inhibit the active site of MMP2.

Results: The virtual screening process has identified five potential candidates: ZINC000000001412, ZINC000001612328, ZINC000001614079, ZINC000000119988, and ZINC0000000047553. These candidates were selected based on their strong binding affinities and interactions with MMP2. These compounds, which adhere to Lipinski's Rule of Five and have significant physicochemical properties, show promise as MMP2 inhibitors.

Conclusion: The finding of this study indicates a preliminary investigation into an innovative approach for managing cancer that inhibits the invasion and dissemination of cancer cells.  

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Exploring Natural Compounds as Promising Matrix Metalloproteinase-2 inhibitors for Cancer Management: A Biocomputational Study
Akram Ahmed Aloqbi, pages 674-678
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 Background: Matrix metalloproteinase-2 (MMP2) plays a role in breaking down the components of the extracellular matrix, which allows cancer cells to advance and invade. Therefore, the inhibition of MMP2 shows potential as a promising strategy for treating cancer.

Methods: This study employed computational screening to identify MMP2 inhibitors from a collection of 2,405 natural compounds. GLXC-26716, the co-crystal ligand of MMP2, served as the positive control. Virtual screening was performed using PyRx 8.0 software to find molecules that might inhibit the active site of MMP2.

Results: The virtual screening process has identified five potential candidates: ZINC000000001412, ZINC000001612328, ZINC000001614079, ZINC000000119988, and ZINC0000000047553. These candidates were selected based on their strong binding affinities and interactions with MMP2. These compounds, which adhere to Lipinski's Rule of Five and have significant physicochemical properties, show promise as MMP2 inhibitors.

Conclusion: The finding of this study indicates a preliminary investigation into an innovative approach for managing cancer that inhibits the invasion and dissemination of cancer cells.  

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Identification of phytochemicals as potential inhibitors of Candida albicans SAP3: An in-silico approach
Zain Alhindi, pages 679-683
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 Background: Candida albicans is a major threat to human health, causing a variety of diseases. Candida albicans can overgrow and trigger infections like thrush, diaper rash, and vaginal yeast infections. Therefore, finding potential inhibitors of C. albicans from plant sources has become important, recently.

Methods: This study investigated the interaction of natural compounds (N = 450) from the ZINC database with the C. albicans SAP3 protein. PyRx tools were used to conduct molecular docking analyses, which included ligand and target receptor preparation, data analysis, and visualization.

Result: The binding energies between the natural compounds and SAP3 ranged from -8.8 to -7.8 kcal/mol, indicating that the interactions were predominantly strong. Among the compounds screened, ZINC000252509014, ZINC000252509722, ZINC000253389136, ZINC000251981944, and ZINC000252480871 were extensively explored in this study because they were found to be the best among others. Furthermore, these compounds showed favorable drug-like properties.

Conclusion: These findings lay the groundwork for future research aimed at developing potent SAP3 inhibitors to treat C. albicans infections. 

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Application of the PCR for diagnosis and monitoring of rotavirus in calves during treatment with the antiviral agent Triazavirin
Tatiana Reshetnikova, Vladimir Kuzmin, Tatyana Krylova, pages 684-688
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 Background: Rotavirus is a significant cause of morbidity and mortality in neonatal calves, leading to severe gastrointestinal and systemic illnesses. Current treatments primarily focus on symptomatic relief and prevention through vaccines, with limited use of antiviral agents. This article  considers the issue of experiments using the antiviral agent Triazavirin in the treatment of rotavirus infection in calves, with tracking the amount of virus during the polymerase chain reaction examination in fecal samples.

Methods: The experiment was conducted at a Mordovian Republican Veterinary Laboratory in the National Research Mordovian State University named after N.P. Ogarev (Republic of Mordovia, Russia), between February and March 2023. Two groups of calves, experimental and control, were assessed. The experimental group received Triazavirin orally at a dose of 500 mg per day for 7 days. Before and after the experiment, fecal samples were taken from experimental calves for the PCR study of the ribonucleic acid of group A rotaviruses.

Results: Treatment with Triazavirin resulted in a reduction of the rotavirus load by an average of 35.2%. The PCR analyses before and after treatment indicated a substantial decrease in viral RNA in the majority of treated calves, with variable individual responses.

Conclusions: Triazavirin demonstrates potential as an effective antiviral treatment for rotavirus in calves, reducing viral load and possibly improving clinical outcomes. The use of PCR provided a reliable method for monitoring viral levels, thereby facilitating effective management of this infectious disease. 

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Exploring the Sequence analysis of the Two-Component System Response Regulator OmpR in Multi-Drug Resistant (MDR) Acinetobacter baumannii
Israa Radwan Ali, Duaa Mohammed Abdulsatar, Ruqaia Sabbar Salman, Anfal Mohammed Khudhair, pages 689-695
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 Background: The study focuses on the clinical profiles, antibiotic susceptibility, and genetic characteristics of 25 A. baumannii isolates from patients with urinary tract infections (UTIs). Patient Cohort: Ages range from 18 to 70 years, with a male to female ratio of 1.375. Notable Finding: Some multidrug-resistant (MDR) isolates had commonalities with extensively drug-resistant (XDR) variations, showing the evolution of drug resistance.

Methods: Antibiotic Efficacy Analysis: Evaluation of antibiotic efficacy when the variable moved from lower (Bin 1) to higher (Bin 2) levels. Correlation Matrix: An analysis of antibiotic correlations to better identify potential cross-resistance and shared characteristics. Genetic Diversity Analysis: An examination of variants in the OmpR gene, including mutations and polymorphisms. Sequence analysis is used to identify point mutations in OmpR, with an emphasis on transitions such as adenine (A) to guanine (G).

Results: Significant improvement in antibiotic efficacy from Bin 1 to Bin 2. Correlation Findings: Antibiotics have complex interactions, which may indicate cross-resistance. Genetic diversity: Variations in the OmpR gene have implications for virulence and adaptability. Sequence Analysis: The majority of point mutations in OmpR were transitions, with A typically changing to G.

Conclusion: In Iraq, researchers have discovered the first evidence that clinical resistance in A. baumannii may be caused by structural alterations in the OmpR gene. A. baumannii isolates' genetic diversity at certain locations suggests possible implications on virulence and adaptation. 

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Use of Biological Agents to Control the Number of Halyomorpha halys Stål
Irina S. Agasyeva, Vladimir Ya. Ismailov, Maria Petrishcheva, Anton S. Nastasiy, pages 696-702
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 Background: Halyomorpha halys Stål, commonly known as the brown marmorated stink bug, poses a significant threat to various crops, necessitating repeated seasonal insecticide applications for control. However, these methods carry environmental and human health risks. Considering these concerns, there is a pressing need to explore alternative, eco-friendly approaches to managing H. halys populations. This study sets out to investigate innovative strategies that minimize reliance on harmful chemicals while still effectively controlling the stink bug menace. By delving into the realm of sustainable pest management, the aim is to develop methods that strike a balance between pest control efficacy and environmental stewardship. This study aims to explore eco-friendly approaches to managing H. halys populations.

Methods: The research involved monitoring the population dynamics of H. halys in the central zone of the Krasnodar Territory, Russia. It also evaluated the efficacy of local parasitic insects Pediobius cassidae and Anastatus bifasciatus in reducing pest infestations, conducted laboratory tests on the insecticidal properties of essential oils from plants such as wormwood, coriander, and Siberian fir, and performed field trials on biological control agents.

Results: Infestation rates by parasitic insects remained relatively low, ranging from 5-10%. Laboratory experiments demonstrated the potent insecticidal properties of essential oils derived from wormwood, coriander, and Siberian fir. Field trials further confirmed significant mortality rates of H. halys specimens when exposed to biological control agents.

Conclusions: The study underscores the potential efficacy of utilizing parasitic insects and plant-derived essential oils in managing H. halys populations. These findings offer practical insights for promoting sustainable agricultural practices in the central zone of the Krasnodar Territory. 

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