Iraqi Journal of Biotechnology

Molecular studies were carried out to estimate genetic variation and genetic distance in
selected genotypes (Furat, 4H, 7H) and two local cultivars (Iraq, Orok) wheat. DNA was extracted from
plant leaves of these genotypes and cultivars at potting stage. The PCR reaction method was used to
estimate the genetic variation. Six primers were used in this study. Data of polymorphic fragment were
analyzed to determine the genetic relationship between the genotypes and cultivars, and also genetic
distance was estimated. Bands differed between cultivars in number and molecular weight which
appeared in these genotypes and cultivars using six primers exhibition genetic variation. Results of the
dendrogram showed that the selected genotypes and cultivars are divided into three groups according to
the genetic variation between them. Results also indicated that the local cultivars and selected genotypes
differed in their genetic distance.

Diarrhea the defined as the condition of having extra stools as normal for that than individual
or as passage of watery stool for more than three times in 24 hours. Giardia lambllia is gastrointestinal
parasite that causes giardiasis, one of the causes of diarrhea in humans .The parasite is distributed
globally, and children are more at risk of infection than adults. This study aimed to determine the
prevalence of G. lamblia in stool samples of diarrheic patients. This study is carried out in Al-Rifea
district in Al-Rifai general Hospital / Thi –Qar Province which included collection of stool samples from
diarrheic patients at a period extended from October / 2017 – January / 2018. 603 stool samples were
taken from patients with different ages to for examined by microscopic examination and PCR technique.
The results showed the percentage of positive samples G. lamblia by microscopic examination was
(8.1%) and negative samples was(91.9%). The highest infected patients found (8.2%) in males and
lowest infected patients found (7.0%) in females. The highest infected patients found (11.8%) in Rural
area and lowest infected patients found (7.2%) in Urban area. According to age group the highest
infected patients was(10.0%) in age group less than (1-10 years) and lowest infected patients found
(4.5%) in age group (21-30) years. while results of PCR from (96) sample were positive in 55 samples
with percentage (57.3)% and 41 negative samples with percentage of (42.7%). The highest infected
patients found (55.5%) in males and lowest infected patients found (59.5%) in females. The highest
infected patients found (64.4%) in rural area and lowest infected patients found (50.9%) in Urban area.
Age group (1-10years) was the highest infected patients which was (81.2%) and lowest infected patients
which was (31-40) years.

Breast cancer is a complex disease encompassing multiple tumor entities, each characterized
by distinct morphology, behavior and clinical implications. Hormone receptor status and HER2 status are
of critical interest in determining the prognosis of breast cancer patients. Their status is routinely assessed
by immunohistochemistry (IHC). The aim of this study is to clarify the differences in the expression of
the established prognostic and predictive markers ER, PR and HER2 with breast cancer by IHC. This
study was conducted at AL-SADER Hospital in AL-Najaf and Medical city in Baghdad, Iraq, from July
2018 to April 2019. It included fresh tissue of 48 patient women with breast cancer.
Immunohistochemical staining with antibodies for Estrogen Receptor (ER), Progesterone Receptor (PR)
and Human Epidermal Growth factor 2 (Her 2) was performed and then breast cancers were classified
into four molecular subtypes: Luminal A (ER/PR +, HER2-), Luminal B (ER/PR +, HER2+), Triple
Negative Breast cancer (Basal like) (ER/PR -, HER2-) and HER 2 (ER/PR -, HER2+). Clinical
parameters were compared using chi-square test. In the current study ER receptors was 72.91% (35/48),
PR positive receptors 56.25 % (27/48) and that 11 out of 48 malignant cases were represent as 22.91 %
were positive for her2/neu expression. Corresponding for the hormones statues, the molecular subtype
was: Luminal A group 56.25% (27/48), Luminal B subtype which was 16.66% (8/48), then a group with
triple negative ( ER-, PR- and HER2- ) was 18.75% (9/48) cases that represent Basal like group, while the
other groups appeared least frequency as a following: HER +, ER- and PR- were (4) cases only which
were referred to HER2 subtype group. There is certain variation among the molecular subtype of breast
cancer with prevalence of certain types among different regional population.

The present study is an attempt for detection of Candida albicans, Candida glabrata,
Candida tropicalis and Candida krusei by conventional methods and multiplex methods as Vitik 2 system
for these Candida species in vaginal swabs that collected from female patients clinically diagnosed with
vulvovaginitis to find out the more specific, and sputum swab collected from male and female patients,
sensitive and rapid method for diagnosis of this yeast. During the period from early December 2018 to the
end May 2019, out of 110 samples a total of 110 vaginal swabs have been collected from female patients
and sputum swab collected from male and female patients attending hospitals in Baghdad city (Al-Karkh
and AL-Samarai Hospitals) and in Tikrit (General Tikrit Hospital) aged from 1-71 years. To sum up 72
out of 110 were detected for Candida species, To sum up 36 out of 72 sputum swabs were detected for C.
albicans, 16 out of 72 sputum swabs were detected for C. glabrata, 7 out of 72 sputum swabs were
detected for C. tropicalis, 10 out of 72 sputum swabs were detected for C. krusi and 3 out of 72 sputum
swabs were detected for negative result.To sum up 38 out of 38 vaginal swabs were detected for C.
albicans. A number of female from summation account 72 patient while summation of male 38 patient
depending on the morphologic characteristics of these Candida species on the culture media including
Sabouraud′s dextrose agar and CHROMagar, as well as the biochemical tests including automated
biochemical tests such as VITEK 2 system. Then we performed a pharmacological sensitivity check for
the following drugs using the Vitk2, Candida albicans which showed sensitivity to Voriconazole with
Minimum inhibition concentration MIC ( 0.12), Flucytosine ( 1), Amphotericin B ( 0.5), Fluconazole (1),
Micafungin (0.06),Caspofungin (0.25).Fluconazole ≤ 1.While, the results to the candida spp. as follows
Fluconazole ≤ 1,voreconazole ≤ 0.12, caspofungin ≤ 0.25,Micafungin ≤ 0.06,Amphotricin B 1and
flucytocine ≤ 1, which sensitive to all antifungals. CHROM agar Candida medium was shown to be
extremely helpful in routine clinical mycology service, facilitating the detection of mixed cultures of
yeasts and allowing directly identification of C. albicans as well as rapid pre summative identification of
the yeast.

Tomato yellow leaf curl virus (TYLCV) is one on the top ten supreme pathogens threatening
tomato (Solanum lycopersicum L.) production worldwide. While TYLCV is a serious disease in tomato
production areas of Iraq under field and plastic tunnels conditions, little is known about genomic features
of these viruses from Iraq. Total DNA was extracted from infected tomatoes were collected from two
growing areas of Iraq. A partial sequence of the TYLCV was carried out, and phylogenetic study was
conducted to understand the genetic diversity of the TYLCV in these regions. Based on the DNA
sequence and phylogenetic analyses, the two collected virus strains were confirmed as TYLCV strains.
The two viral isolates that have been characterized in this study were shared 98.91% and 98.38 %
nucleotide sequence identity with a previously characterized Iraqi and Lebanese TYLCV isolates,
(JQ35499.1and EF051116.1) respectively. The result showed that Iraqi isolates shared highest sequence
identity (<98 %) with group of TYLCV complete genome sequencing from Iran, Jordan, Lebanon and
Israel include viral strains JQ231214.1, EF054893.1, AB116631.1, AY594174.1, AB110217.1,
AB116629.1, AB116630.1 and AB116631.1.

Oligozoospermia is a type of male infertility in which the number of sperms is under 15
million/ml, it is caused by different factors but it is mostly associated with idiopathic disorders. MLH1 is
a part of a genes system which codes for proteins responsible mainly for correcting post replicative
DNA errors.rs1800734 is a single nucleotide polymorphism lies in the MLH1 CpG island at -93 from the
translation start site while rs4647269 is an intron variant that lies in intron 9 . ). Blood and semen samples
were collected from Baghdad Specialist Fertility Centre through the period from 1st of August 2018 until
the end of February 2019 and the study was carried out in the Laboratories of Institute of Genetic
Engineering and Biotechnology for Postgraduate Studies - University of Baghdad .Genotyping of MLH1
gene for SNPs rs1800743 and rs4647269 was done using Taqman genotyping assay by RT-PCR.
Hormonal analysis was performed by using fluorescence immunoassay (FIA) on AFIAS autoanalyzer
.The study showed that There were no significant differences between healthy and oligozoospermia
patients in any of the genotypes of the SNP rs1800734, hence the SNP have no connection to the
infertility incidence. As for rs4647269, The results showed that the CC genotype of rs4647269 was
significantly higher ( p<0.05) in healthy patient (15(30%) versus 9 (18%) in oligozoospermia patient
contrary to TT genotype that was significantly higher in oligozoospermia patients15 (30%) versus
7(14%).The heterozygous genotype CT had no significance between the two groups.Seminal fluid
analysis showed a significant( p<0.05) decrease in sperm count (9.65 ± 0.89) ,motility(24.34 ± 2.072)
,activity8.06 ± 1.15 and normal sperms(4.60 ± 0.66) in the oligozoospermia patients .In oligozoospermia
patients, mutant genotype TT of the SNP rs4647269showed a significance increase (P<0.05), compared to
the wild genotype CC. Effect of SNP rs4647269 on FSH appear in the wild and heterozygous genotypes
as an increase in oligozoospermia patients compared to the healthy ones.In rs4647269 there was a
significant increase (P<0.05) in prolactin in both the wild and the mutant genotypes of the
oligozoospermia patients compared to the control, only the heterozygous resulted in a significant (P<0.05)
decrease in the oligozoospermia patients.only the wild genotype CC showed a significant decrease
(P<0.05) in testosterone concentration of oligozoospermia patients.the results of rs1800734 showed that
the only genotype that showed a significant difference in LH levels between oligozoospermia patients
and control is GA genotype (P<0.05). FSH results were significantly higher (P<0.01) in oligozoospermia
patients of the genotype GG than control of the same genotype.. The results show a significant increase
(P<0.01) in PRL levels that accompanied the GG genotype in oligozoospermia patient. While the mutant
genotype AA resulted in a significant decrease (P<0.05) in oligozoospermia patients. Only GG genotypes
(the wild genotypes) showed a significant decrease (P<0.05) in testosterone concentration of
oligozoospermia patients . This study have observed 4 possible haplotypes, GC hablotype was the most
common in both groups (patients and controls) and it was significantly higher( p<0.05) in control. Also
the GT hablotype was higher( p<0.05) in oligozoospermia patients than healthy controls. 5 of 8 possible
haplotype combinations showed no statistically significant differences between oligozoospermia patients
and control groups.This study concluded that 1. Both rs1800734 and rs4647269 SNPs of MLH1 gene
have no role in the incidence of oligozoospermia in Iraqi men.

Cancer is the major health issue around the world, in spite of the development in the diagnosis
and therapy methods. The discovery of new anticancer drugs from plants become the new trend in the
scientific research. The phytochemicals, consider selective in their action and specially not to be harmful for
normal cell like other cancer treatments. In the current study, leaves of Bauhinia variegata were collected
from Baghdad University garden in spring of 2019 and Glucokinin was isolated and purified from the
methanol extract of B. variegata by using column chromatography. The cytotoxicity effects of Glucokinin
were investigated via MTT assay against cancer cell lines HepG2, A549 and WRL normal cell line at 24, 48
and 72h. by using different concentrations. The result showed that, the highest cytotoxicity of Glucokinin at
400 µg/ml was 46.53% (std. ±5.8) at 72h. against A549 cell line. And the half maximal inhibitory
concentration (IC50) of Glucokinin that inhibit the A549 cancer cell line was 114.5 µg/ml at 72h. The
greatest cytotoxicity effect against HepG2 was 71.75% (std.±6.5) at 72h. The (IC50) of Glucokinin that
inhibit the proliferation of HepG2 cancer cell line was 51.24 µg/ml at 72h. Additionally, Glucokinin showed
high IC50 against WRL normal cell line. In conclusion, these finding show the probability of using
Glucokinin as anticancer agent.