Iraqi Journal of Biotechnology

Resistance of the acute myeloid leukemia (AML) to chemotherapy is a major clinical
problem resulting in poor patients’ prognosis. One of the known multidrug resistance (MDR) mechanisms
in AML is the overexpression of efflux pumps belonging to the superfamily of ATP- Binding Cassette
(ABC) transporters such as ABCB1 and ABCG2. For this purpose, the present study investigated the gene
expression of these genes using real-time PCR. Blood samples of AML Iraqi patients were collected at
diagnosis and after induction therapy to investigate the correlation of ABC genes expression and response
to chemotherapy. Results reveled low expression of ABCB1 and ABCG2 at diagnosis when compared
with those after treatment and healthy subjects suggesting that the gene expression of these genes could
be considered as a marker of AML regression.

Staphylococcus aureus is associated with multi drug resistance infections and high levels of
illness and the efflux pump has vital role in multi drug resistance for antimicrobial agent in S. aureus. (one
hundred and fifteen) clinical specimens (burns, blood, Ear, Nasal, Urinary tract infection and wounds) were
collected from patients attending different hospitals in Baghdad. S. aureus has been identified using
biochemical,Vitek-2 system Furthermore, molecular methods. Molecular method was depended on The
polymerase chain reaction (PCR) assay was applied to determine the major specific genes, 16S rRNA and
medA gene were used to diagnos Staphylococcus genus and med A gene to diagnos S. aureus species. Out of
115 isolates, 55 were identified by Vitek-2 system the result about 20 S. aureus isolate Furthermore
identified S. aureus by PCR Just to make sure. all S. aureus isolates were tested toward the different class of
clinically important antibiotics by using vitek 2 system. The results of resistance were as following:
Penicillin G (10) unit (100%) Oxacillin (99%), Imipenem (3%). while the resistance to chloramphenicol,
(27%), Azithromycin (04%), Erythromycin was (33%), Tetracycline (72%) Doxycycline is (7%) ,
Ceftazidime (99%), Ciprofloxacin (72%). The aim of this study is to investigate the prevalence of number of
chromosomal efflux pumps genes). The results indicated the presence of 16s rRNA gene in all isolates (100
%) (from 20 isolates) The med A gene has been recorded the second highest prevalence present in 99
(92.71%) isolates., these results showed the role of 16S rRNA for molecular detection Staphylococcus genus
and using medA gene in S. aureus bacteria considered one of the most important features in the description of
Staphylococcus aureus isolates.

Staphylococcus aureus is an opportunistic pathogen. It is most common in skin and soft tissue.
This microbe can cause more diseases as a burn inflammation and tonsillitis through the production of
virulence factors. In this study, one hundred and fifty (051) samples were collected from patients with
inflammation from different clinical sources and different age groups during the period from December 2020
to the end of April 2021.from main hospitals in Baghdad. All isolates were diagnosed based on microscopic
examinations and morphological characteristics using Mannitol salt agar, Blood agar in addition to
biochemical tests.The diagnosis was confirmed through Vitek system and API staph kit. Antibiotic
susceptibility testing was performed on all identified isolates, and the findings revealed that 12 isolates
VRSA.DNA was extracted from bacterial isolates and sepreated by electrophoresis to measure DNA purity
and integrity. Molecular diagnosis of bacteria was done using a polymerase chain reaction based on the
16SrRNA gene and the results were (100%) as S. aureus. In addition the individual interactions polymerized
were used to determine the vancomycin resistance genes. It was found that all isolates were vancomycin
resistance through the presence of the van A gene, while the results of the presence of genes van B were
(66.6%).

Increasing the rate of antibiotic-resistant pathogens causing serious infections and death has
become a global health threat. However, the synthesis of antibiotics and new drugs is expensive and
undertaken through multi phases and clinical trials. Therefore, repurposing old drugs to treat antibiotic
resistant bacteria is a good alternative way. Escherichia coli (E.coli) is a common pathogen of urinary tract
infection, and its capability to form bifilms further contributes to its virulence and antibiotic resistance, posing
a big crisis to global public health. Uropathogenic E.coli were isolated then tested by Vitek2 and disk
diffusion method to know which antibiotics are resistant to E.coli. Then, real-time PCR, conventional PCR,
and genes sequences tools were used to identify the strain of isolates E.coli. To do repurposing antibacterial
drugs or to find a new compound, we performed a virtual screening on Pharrmit provides an online,
interactive environment for the virtual screening of large compound databases using pharmacophores,
molecular shape, and energy minimization to predict a compound that has biological activity against the
uropathogenic E. coli. A total of 14678 small molecules from the Drug Bank database were subjected to
screening via Pharmit. Nine potential hits were obtained (100009383 - 1531009). Molecular docking can be
used to evaluate the affinity of the new compounds to predict the binding affinity to FimH receptor.

An experiment was conducted in a greenhouse, located in the College of agriculture, Wasit
University, Iraq. Tomato was planted and grown in winter season (November 2020 - April, 2021). The
experiment aimed to study the effect of Trichoderma and nitrogen fertilizer on the growth and yield of
tomato. Treatments comprised no application treatment as a control (T1), 100 kg N / h-1 application ( T2),
160 kg N / h-1 application (T3), 200 kg N / h-1 application (T4), 0 kg N / h-1 application with 100 g of
Trichoderma (T5), 100 kg N / h-1 application with 100 g of Trichoderma (T6), 160 kg N / h-1 application with
100 g of Trichoderma (T7) and, 200 kg N / h-1 application with 100 g of Trichoderma (T8). A randomized
completely block design (RCBD) was used with three replicates. Treatment (T6) showed a significant
superiority .in plant height (cm), leaf area (dm2. Plant-1), leaves number (leaf. Plant-1), leaf chlorophyll content
(SPAD), Plant dry weight (g), fruit wei

Sumac (Rhus coriaria L.) is one of the popular spices used in different medical purposes,
different studies show that sumac fruits, leaves and seeds contained different types of flavonoids, so the
present work was aimed to investigate quantitatively and qualitatively the polyphenolic components of
sumac fruits using HPLC and different chemical methods using different standards and modified
chemical methods as Ciocalteu phenol reagent and other reagents to do this project of evaluate
quantitatively the total phenolic contents TFC and total flavonoids contents TFC in different Sumac
extracted solutions using different organic solvents (Ethanol, Methanol and Ethyl Acetate) of different
aqueous dilutions, while the total flavonoids evaluated qualitatively using HPLC were about 8 types of
flavonoids have been identified in different aqueous methanolic concentrations (50, 75 and 95%v/v) and
the higher methanolic concentration extracts shows higher extracted quantities of polyphenolic
compounds and flavonoids in compare with the other aqueous diluted methanolic extraction solvents.

The present study including collection of 100 serum samples and from totally100 patients
include 28 males and 72 females clinically diagnosed with type 2 diabetes mellitus from binging of July
to end of August 2016, who they attended to Al-Zafaraniyah Hospital. The estimation of fasting blood
sugar (FBS) concentrations in type 2 diabetes mellitus patients, have indicated that the FBS
concentrations could be divided into three groups; the first group contains 85 males and females diabetic
patients with FBS concentrations range from 120-200 mg/dl and the second group contains 14 males and
females diabetic patients with FBS concentrations range from 200-300 mg/dl, whereas the third group
contains one male diabetic patients with FBS concentration more than 300 mg/dl in both of males and
females diabetic patients. The determination of ABO blood groups and Rh in type 2 diabetes mellitus
patients, have indicated that the blood group A was the most dominant blood group comparing to other
blood groups followed by blood groups B, O and finally AB. The results showed that to sum up 39 out of
100 blood samples were determined for blood group A, 28 of blood samples were determined for blood
group B, 24 of blood samples were determined for blood group O and 9 of blood samples were
determined for blood group AB from both of males and females patients clinically diagnosed with type 2
diabetes mellitus.

Male infertility is a complicated problem with a strong genetic and epigenetic foundation.
Imprinted genes survive fertilization and retain their parent-specific germline patterns, while aberrant
methylation imprints can be passed directly from the father's sperm to the developing embryo. Semen
samples were collected from patients (number=100) (50 infertile oligospermia men and 50 fertile
normospermia men) for epigenetic study (methylation pattern) after DNA isolation from seminal fluid
samples using a kit for extraction DNA and converted to methylation pattern, then analyzed methylated
DNA was used quantitative real-time polymerase chain reaction by high-resolution melting technique
for H19, SNRPN, and LINE-1 genes. Also, semen samples were used for the molecular study (gene
expression) after RNA was extracted using a kit of extraction and converted to complementary DNA, then
used quantitative reverse transcription real-time polymerase chain reaction for H19, SNRPN, and LINE
1 genes to determine expression levels. The current study was present for DNA methylation was significant
in the infertile group compared with fertile group, the defects on gene promoters and the affect on gene
expression may serve as epigenomic biomarkers for the assessment of oligospermia infertility in men with
multiple sperm defects.

Urinary tract infections (UTIs) are one of the main health problems caused by various
microorganisms including uropathogenic Escherichia coli (UPEC). UPEC strains are the most frequent
pathogens responsible for 85% and 50% of community and hospital acquired UTIs, respectively. UPEC
strains have specific virulence factors that can result in worsening of UTIs, including adhesion trait. In
this study, Fifty-four UPEC isolates were collected from patients with various clinical manifestations of
UTIs attending different hospitals in Baghdad/Iraq from November 2020 to March 2021. These isolates
were identified by culturing on differential media MacConkey agar and selective media Hardy chromo
UTI agar in addition to the blood agar, then confirmed the identification using Vitek 2 automated system.
The prevalence of fimbrial adhesion related gene (fimH) and non fimbrial adhesion related gene (tosA)
gene were screened among 45 UPEC isolats using polymerase chain reaction assay with specific primers.
The result reported the presence of studied genes was in high percentage (95.6%) for fimH coded gene
while was in moderate percentage (47%) for tosA coded gene. thus, fimH and tosA could be used as a
possible diagnostic marker in addition to indicate the level of UPEC pathogenicity.

The protein tyrosine phosphatase nonreceptor 22 (PTPN22) is associated with susceptibility
to group of autoimmune diseases. The functional polymorphism in PTPN22 at 1857 It is considered a
risk factor for vitiligo susceptibility in European; however, controversy exits in other populations.
Present study aimed to study The exon 14 region of the PTPN22,and determine whether the PTPN22
C1857T polymorphism confers susceptibility to vitiligo in Iraqi patients. Genomic DNA was extracted
and amplified using polymerase chain reaction and sanger sequencing method. The frequency of the CC
and CT genotypes were 97.30% (36) and 2.70% (1) in the generalized vitiligo patients, respectively, and
93.88% (46) and 6.12% (3) in the healthy controls, respectively. There was no statistically significant
difference between the generalized vitiligo patients and healthy controls.The homozygotes genotype (TT)
was absent in both the generalized vitiligo patients and healthy controls. there was no significant
difference between two groups. The frequencies of the polymorphic T allele were 1.28% and 3.06% in the
patient and the control groups. The genotype and allele frequency of the rs2476601 did not show any
statistical significance (p > 0.05) in its association with generalized vitiligo.

The increasing incidence of carbapenems-resistant K. pneumoniae has been considered a public
health problem especially among patients with urinary tract infections (UTIs) . Objective: The aim of this
study was to evaluate the frequency of carbapenemase genes in K. pneumoniae isolates from UTIs and their
role in the antibiotics resistance rate of these isolates. Material and methods: Eighty K. pneumoniae isolates
were obtained from 250 urine samples (November 2020 to march 2021) from Baghdad hospitals, Iraq.
Antimicrobial susceptibility was assessed by disc diffusion method. The carbapenemase genes blaOXA48,
blaVIM, blaIMP, blaKPC and blaNDM were investigated by polymerase chain reaction (PCR). Results: K.
pneumoniae isolates were detected in 32% of the tested samples in the patients with UTIs, and the prevalence
of the isolates were higher in females in comparison with males. The results showed high resistance rates to
Amikacin and Rifambicin (97.5%), to Cefotaxime, Ceftazidime and Piperacillin (92.5%), followed by
Ceftriaxone and Aztreonam (87.5%), Cefepime (85%), Imipenem (62.5%), Meropenem and Gentamicin
(50%). The most prevalent genes were blaIMP (37.5%), blaOXA48 (30%), and blaVIM (28.75%), followed
by blaKPC (20%), and blaNDM (11.25%). The most isolates which harbor the carbapenemase genes revealed
a high resistance to cephalosporins and carbapenems. Conclusion: The rate of multi-drug and carbapenems
resistance was high in the K. pneumoniae isolates from UTIs in Baghdad hospitals. Colistin could be the drug
of choice for the treatments of carbapenemase-producing K. pneumoniae. Our study provides a better
understanding of the carbapenemase genes distribution among the local isolates in UTIs.

β-thalassemia major is a genetic disorder of hemoglobin production that results in a diminished
rate of synthesis of one or more of the globin chains causing variable degrees of anemia. This study was
conducted on 90 β-thalassemia patients and 60 healthy as a control group. Blood samples were obtained
from Wasit Center for Hereditary Anemia through the period from August 2020 to January 2021. The
study was conducted to evaluate the serum immunological markers (TLR-3, TLR4) and (IL-7, IL37) in
Iraqi β-thalassemia patients. The study was approved by the council of the Institute of Genetic
Engineering and Biotechnology for the Postgraduate Studies / University of Baghdad. Human TLR-3,
TLR-4, IL-7, and IL-37 concentrations were estimated by ELISA. Results appeared that there was a
significant increase in the level of TLR-3 and TLR-4 in the serum of β-thalassemia patients as well as a
significant increase in the level of IL-7 in β-thalassemia patients (221.34±6.11 pg/ml) compared with the
control (171.96±4.77 pg/ml). Whereas a non-significant difference in the level of IL-37 was recorded in
patients and the control group. This study suggested the presence of an imbalanced immune condition
involving inflammation and immunosuppression in thalassemia patients.