Iraqi Journal of Biotechnology

High blood pressure is defined as a systolic blood pressure of 140 mm Hg or more, or a
diastolic blood pressure of 90 mm Hg or more. Blood pressure is the force exerted by the circulation of
blood on the walls of the body's arteries, which are the main blood vessels in the body. High blood
pressure is classified into normal, first stage, second stage, or third stage. Risk factors include lifestyle
factors, environmental factors, and genetics. There are two types of high blood pressure, essential
hypertension and non-essential hypertension. Blood samples were collected, representing 50 samples
from patients with high blood pressure, in addition to 50 healthy samples. This study was conducted with
the aim of studying the relationship between genetic variation of some genes and environmental factors
associated with essential blood pressure in Iraqi patients. The studies were conducted in the laboratories
of the Institute of Genetic Engineering - University of Baghdad, as well as in the research laboratories of
the Ministry of Science and Technology. Total genomic DNA was extracted using a special kit from fresh
unfrozen blood samples, and then normal polymerase chain reaction and PCR-RFLP were used to detect
mutations in Cluster of differentiation 36 rs1761667 G>A gene using primers and specialized restriction
enzyme is HhaI. The results of the GG wild-type Cluster of differentiation 36 gene showed a band (161+
264 bp), while the heterozygous GA genotypes showed (161, 264, and 425 bp), respectively, and for the
mutant AA genotypes (425 bp). Nitrogenous base sequence analysis (Sequencing) was conducted for both
infected and healthy samples. The results of the study showed that the incidence of hypertension in the
age group (20-65) years was higher in males than in females, reaching (60% and 40%), respectively. The
study showed that stress, smoking and exposure to pollution all have a clear and dangerous effect on the
percentage of people with high blood pressure. The results also showed that mutations between patients
and apparently healthy people in the Cluster of differentiation 36 gene may cause an increase in blood
pressure for the wild-type GG is (56%) versus (44%) a significant level (p<0.05), while for the
heterozygous GA it is (20%) versus (24%) ) found not significant, either for the homozygous mutant AA
is (24%) versus (32%) with a level (p<0.05). These results indicate that genetic variation (G>A) may be a
risk factor (GG)) for high blood pressure.

Soil contaminated by petroleum hydrocarbons (PHCs) is a global severe environmental
problem, especially if we consider that Iraq has been subjected to several wars and the widespread use of
generators to provide electrical energy. In addition to oil extraction sites, gas manufacturing sites,
refineries, and ammunition waste sites. Therefore, the need to know the best ways to treat these polluted
soils has been increased. For this purpose, this study was conducted to isolate bacteria and fungi from
contaminated and non-polluting soils that synergism with plant for remediation of oil-contaminated soil.
The study was carried out in the plastic shade scale, and seeds were cultivated in plastic pots filled with
12 kg of soil. The experiment was laid out in Completely Randomized Block Design (CRBD) with three
replications. Each replication comprised the following treatments randomly: AL-Daura power station soil
planted with maize, AL-Daura oil refinery soil planted with maize, AL-Daura oil refinery soil unplanted
(abiotic stress), AL-Daura oil refinery soil sterilized unplanted (treated with AgNO3 (biotic stress)), in
addition to control soil planted with maize. The total number of bacteria and fungi and hydrocarbon
(HCs)-degrading microorganisms were evaluated during phytoremediation by cultured in a proper media
agar. The rhizospheric bacterial and fungal count of the planted soil increased at the end of the
experiment. Totally, the study obtained 11 pure bacterial isolates and 7 pure fungal isolates that could
grow on Bushnell Hass (BH) agar medium containing crude oil. Current study suggested that the used of
maize plants played an important role in stimulation the microorganisms. They were increased the
number of microorganisms. Also, the petroleum degraders microbial increased in the oil-contaminated
soil and the non-contaminated soil. The microbial degraders are found in the entire treatments.

Invasive ductal carcinoma (IDC) is a common breast cancer especially in women. Several factors
may involve in establishment of the breast cancer in general. Variant viruses have been investigated in for
their role in the establishment of breast cancer, among them is Epstein Barr virus which has been reported in
the patients diagnosed with invasive ductal carcinoma. The study aimed to detect the presence of Epstein–Barr
virus latent membrane protein 1 (LMP1) in a group of Iraqi women diagnosed with IDC and to correlate
expression patterns with the age of the participants and with the grade and stage of cancer. A total of 30
formalin-fixed paraffin-embedded tissue of IDC biopsy samples and 29 formalin-fixed paraffin-embedded
tissues of fibroadenoma as controls were collected to detect the presence of EBV LMP1 using
immunohistochemistry. LMP1 expression was found in 76.67% of cases and in 58.62% of the control
specimens. The expression pattern of LMP1 was determined to be statistically significant (P=0.0037) when
correlated with age, grade and stage. As well as immunohistochemistry was found to be a sensitive technique
in the detection of EBV. Conclusions: The study reveals that the LMP1 was over expressed in our population
cohort.

Male infertility in Iraq, particularly in the previous three decades diagnosed remarkably at a
young age, due to a variety of variables, the most important are war and devastating civil wars, stress,
lifestyle, smoking, dietary habits, and genetic susceptibility. The methylene tetrahydrofolate reductase
(MTHFR) gene has received a lot of interest, mainly because the MTHFR C677T variant was found to be a
risk factor for male infertility, especially in Severe Oligoasthenozoospermia patients. The study aimed to
determine the prevalence of this crucial gene mutation across the country and the estimation of total
endogenic human 8-hydroxyl-2'-deoxyguanosine concentrations as a specific marker of oxidative injury to
sperm DNA. Semen analysis was done for 100 men. The analysis was accounted as recommended by WHO
criteria. All DNA samples successfully extracted using the organic method. The genetic variation of the SNP
MTHFR gene 677 C > T (A222V, rs1801133) was discovered using real-time polymerase chain reaction (RT
PCR) to detect different genotypes. Enzyme-linked immunosorbent assay was performed for measurements
Hydroxydeoxyguanosine (8-OHdG) in semen samples, which is considered as a biomarker of cellular
oxidative stress. The MTHFR C677T gene polymorphisms were found to be a risk factor for male factor
infertility in the Iraqi population in this investigation. Furthermore, this study found a considerably greater
amount of 8-OHdG in sperms from all infertile groups when compared to controls, implying that oxidative
DNA damage has a major impact on sperm characteristics including concentration and progressive motility.

Several studies refer to higher levels of oxidative stress are associated with infertility. The
purpose of this study to know the genotype frequency of GSTM1 and GSTT1 genes in infertility patients
and assest the relationship between the genes and some physiological parameters in infertility patients.
Genotype (GSTM1 and GSTT1) genes were determined by multiplex polymerase chain reaction (PCR).
The result showed a significant increase in serum malondialdehyde (MDA), in unexplained Infertile
(oligozoospermia and asthenozoospermia) groups compared fertile (Control) groups,. While,the result
showed a significant decrease in serum glutathione (GSH) , in unexplained Infertile (oligozoospermia and
asthenozoospermia) groups compared fertile(Control) groups, the percentage of genetic deletion in
oligozoospermia samples as follows : The deletion of GSTM1 ,GSTT1 and null genotype (deletion of
GSTM1 and GSTT1) are most common in infertility patients (oligozoospermia , asthenozoospermia) when
compared to control group.. It could be concluded there use of measuring GSH and MDA activity to
assess infertility patients as the first line deface. These parameters of the marker in infertility patients.

A total of 200 samples of wheat were collected during the period from the beginning of
October 2019 until the end of January 2020 from government and private grain stores and local markets
in Karbala Governorate to determine the percentage of infection with Aspergillus flavus. Molecular
diagnosis of A. flavus was performed using polymerase chain reaction (PCR) technology using the
AFO gene. The results showed 138 samples of the total 200 samples were infected with different fungi.
62 samples of them belong to the Species A .flavus. Of the 62 samples, 23 were strong production, 15
were medium production, and 24 were weak producer of aflatoxin, depending on the Ammonia vapor
test.

The prevalence of gastritis is still elevated all over the world. It is the leading cause of many
diseases like atrophic gastritis, peptic ulcer, and adenocarcinoma of the stomach. The current study aimed
to assess the risk factors in a sample of Iraqi patients was complaining from gastric symptoms and
gastritis. The cross-sectional study was conducted from December 2019 to April 2020. Sixty samples
were collected from both patients and apparently healthy group. The demographic characteristics for each
group include: age, sex, body mass index, diet habits, smoking history, habitual drinking and drug
history. Serums were collected from both groups and anti-Helicobacter pylori IgG were done using
Enzyme Linked ImmunoSorbent Assay (ELISA). The results revealed that the percentage of those within
the age group 30 -50 years old were higher in patients’ group. Smoking, spicy food, abuse of Non-Steroid
Anti-Inflammatory Drugs and H. pylori infection were increased risk of gastritis in a sample of Iraqi
patients.

This study included collecting (120) clinical specimen from different sources, (43) isolates
successfully were diagnosed as P. aeruginosa, representing 35.8% of total isolates. The test for the
antibiotics susceptibility of the bacteria to (10) types of antibiotics showed a clear variation in their
resistance to antibiotics, as all isolates showed resistance (100%) against to cephalexin, (90 %) against to
Gentamicin and 82% against to Ceftazidime,14 isolate out of 43 isolate had MDR. While the results of
the phenotypic detection of pyocin production to resistance isolate (14 isolate) showed the presence of 12
isolates (85.7%), The third screening stage molecular detection of the pyocin S5 gene, which is one of
responsible for the production of Pyocin S5, It was shown that 6 isolates (50%) possess this gene, while 6
isolates (50%), they did not possess this gene.

Lipases are enzymes that have numerous applications in many industries like leather, soaps and
detergents, pharmaceuticals, biofuel, food, textile, etc. Many organisms like plants, fungi, bacteria are known to
produce lipase. The present study was aimed at isolating lipase producing microorganisms from different water
samples which are rich in lipid content like oil mills. As oil mill samples are rich in lipid and fatty acid content it
makes a very good source to find microorganisms capable of degrading lipids by producing lipase enzymes. In
the present study the isolates were identified by morphological, biochemical and molecular characterization.
Purification results of lipase exposed prominent specific activity of 248.4 U/mg, purification fold of 75.50, and
47.7% yield. The purified lipase demonstrated outstanding activity and stability in a temperature range of 25-45
°C and pH (5–10), revealing optimal activity at 35 °C and pH 7. The molecular weight of the enzyme was
estimated to be 63 kDa. Compared to control, the lipase activity was promoted in the presence of calcium
chloride.

A total of 135 soil samples were collected during the period between December 201) to
January 2020. Twenty of them were identified as S. marcescens according to 16S rRNA gene sequence
analysis. The promising characterized and identified isolate was selected for further screening and
evaluation of wheat growth in pots experiment. S marcescens isolates were applied alone and in
combination with varying levels of salt (0, 100, 200, and 300 mM NaCl) to determine its tolerance to
salinity and its role to mitigate the effect of salt on wheat growth, activities of enzymatic antioxidants, and
oxidative damage. In vitro tests showed that this bacterium was able to fix nitrogen, solubilize
phosphorus, production of siderophores, and synthesize indole acetic acid. The presence of the gene for
ACC deaminase production was confirmed using the DegACC primer designed to amplify the accdS
(ACC deaminase) gene. The AH-20 isolate showed growth at high salt (NaCl) concentration of up to
11%, indicating its potential to survive and interact with plants growing in the saline soil. The inoculation
by S. marcescens significantly promoted the growth of wheat plants under salt stress (100–300 mM), and
generated changes in antioxidative enzyme activities (Superoxide dismutase, Peroxidase, and Catalase)
under different salt levels, reducing salinity-induced oxidative damage to the plants.

Klebsiella pneumoniae is one of the main pathogens which cause wound infections.
Biofilm-producers of these bacteria have a high level of resistance to antibiotics and this leads to
complications for the treatment of several infections. The study tested the effect of
ethylenediaminetetraacetic acid (EDTA) on the biofilm formation by multidrug-resistant (MDR),
strong biofilm producer K. pneumoniae isolates from Baghdad hospitals, Iraq. The Minimum Inhibitory
Concentrations (MICs) by Microtiter Plate Assay with resazurin dye and the ability for in vitro biofilm
formation by Microtiter plate assay using crystal violet were detected in MDR K. pneumoniae isolates
in the presence of eight concentrations of EDTA (4 to 512 µg/ml). Out of 45 K. pneumoniae isolates,
35 (77.7 %) were Multi-Drug Resistant (MDR) and 25 (55.5%) were strong biofilm producers. It was
found that all isolates of K. pneumoniae (100 %) were resistant to Ampicillin and Cephalexin, while
these isolates exhibited a low-level resistance against Tigecycline, Meropenem and Imipenem. The
results of Minimum Inhibitory Concentrations revealed that the effect of EDTA on the growth of K.
pneumoniae isolates was recorded at concentrations (32-512 μg/ml). The highest antibiofilm activity by
EDTA was demonstrated at the subinhibitory concentration (256 μg/ml) with biofilm eradication
percent (94.28%), while at very low concentrations (8 μg/ml), it was found an obvious eradication
effect on biofilm (82.11 %). The study suggests that EDTA plays an important role in the early stage of
biofilm formation with a clear effect on the growth of MDR K. pneumoniae.

Psoriasis is a chronic inflammatory disease characterized by a number of immune response
abnormalities. Cytokines are small, biologically active proteins that regulate cell growth, function, and
differentiation and aid in the direction of the immune response and inflammation in psoriasis. This study
was conducted to evaluate serum IL-17A, IL-18, and IL-22, levels by the enzyme-linked immunosorbent
assay (ELISA) for detection the importance and effects these cytokines in psoriasis. Seventy-five
patients with psoriasis (38 females and 37 males) were recruited from the Al Kindi General Teaching
Hospital during the period between the 1st of December 2019 until December 2020, with mean age
(35.30 ±8.64) years old, and 75 apparently healthy volunteers (40 females and 35 males), with mean age
(33.40 ±6.25) years old, with no symptoms or history of psoriasis or other allergic or skin disorders
(control group), In this study 3 ml sera samples from each individual of both groups were collected. The
results demonstrated significant elevation of Serum IL-17A, IL-18, and IL-22 in the patient's group
compared to healthy controls (P≤0.01) in patients with psoriasis compared to healthy control. These data
support the view that serum IL-17A, IL-18, and IL-22 are involved in the pathogenesis of psoriasis,
possibly by induction and maintenance of psoriatic lesions. It recommends the use of cytokine (IL-17A,
IL-18, and IL-22) as a useful follow-up marker for monitoring psoriatic patients and optimizing
therapeutic strategies.