ali malekasgar

چکیده
در NO نقش دارد . علیرغم اثرات مفید (NO) آنزیمی است که در تولید نیتریک اکسید (NOS) زمینه و هدف : نیتریک اکسید سنتاز
و نیتروزیلاسیون پروتئین ها، داراي اثرات سمی (RNS) سیستم هاي مختلف بدن، تولید بیش از حد آن به خاطر تولید گونه هاي نیتروژن فعال
بر مهار فعالیت نیتریک (L-NMMA) و ان جی- مونو متیل آرژینین (ADMA) است. این مطالعه به منظور تعیین اثر دي متیل آرژینین نامتقارن
اکسید سنتاز انجام شد.
طی مراحل هموژنیزاسیون، رسوب با سولفات آمونیوم اشباع و کروماتوگرافی NOS روش بررسی : در این مطالعه آزمایشگاهی آنزیم
2 از 500 گرم بافت تازه کلیه گوسفند جدا گردید. طی مراحل جداسازي میزان ',5'-ADP-agarose و DEAE-32 Cellulose روي ستون هاي
در غلظت L-NMMA و ADMA به روش گریس اندازه گیري شد و اثرات مهار کنندگ ی NOS پروتئین به روش برادفورد و فعالیت آنزیم
تعیین گردید. NOS 25 میکرومولار بر فعالیت
0 درصد به دست آمد. وزن ملکولی آنزیم / 0 واحد بر میلی گرم پروتئین و 9 / به ترتیب 6 NOS یافته ها : فعالیت ویژه و بازده خالص سازي
در غلظت هاي 25 میکرو مولار موجب کاهش فعالیت آنزیم به ترتیب L-NMMA و ADMA . 54 حاصل شد KD ، SDS-PAGE با استفاده از
5 میکرومول ، / به ترتیب 32 L-NMMA و ADMA در غیاب و در حضور NOS آنزیم Km 61 درصد گردید. میزان / در حدود 76 درصد و 2
در غیاب و یا در حضور این ترکیبات NOS آنزیم Vmax به دست آمد و در (P<0/ 14 میکرومول ( 001 / و 29 (P<0/ 31/25 میکرومول ( 001
تغییري ایجاد نشد.
داراي L-NMMA در مقایسه با ADMA بوده و NOS داراي اثر بازدارندگی رقابتی بر فعالیت آنزیم L-NMMA و ADMA : نتیجه گیري
قدرت بازدارندگی بیشتري است.
کلید واژه ها : نیتریک اکسید سنتاز ، دي متیل آرژینین نامتقارن ، ان جی- مونو متیل آرژینین ، کلیه
_ __________________________

Abstract
Lipoxygenase (LOX) catalyzes irreversible transfer of oxygen molecule to Arachidonic and Linoleic acid to produce 13 Hydroproxy Octadecadienoic acid. Recent studies showed that the involvement of Lipoxygenase products, leukotrienes, in inflammations and Lipoxygenase pathways acts as mediators of early inflammatory events in atherosclerosis. The aim of the present study is purification and characterization of Lipoxigenase from Human placental.For this aim, the human placental Lipoxigenase was extracted and purified by normal butanol, acetone, ammonium sulphate (30-80%), and gel permeation chromatography on Sephadex G-150. After purification and characterization of LOX, the in vitro inhibitory effect of KCN, NaN3 and some selected bivalent ions such as Co2+, Ni2+, Cu2+, and Zn2+ were checked on the activity of purified LOX. Results showed that specific activity was 123.16 u/mg proteins and the yield of purification was 21.84 percent. Also, it was found that Co2+, Ni2+, KCN, and NaN3 at concentration of 20 mM had inhibitory effect on LOX activity and their inhibitory was 72.4, 58.2, 56.5 and 42.3% respectively. However, Cu2+ stimulated the lipoxygenase activity at the same concentration whereas Zn2+ has no significant effect on LOX activity. With respect to increase of LOX activity in the patient with cardiovascular diseases, Alzheimer disease, cancer, chronic obstructive pulmonary disease (COPD), artherogenesis, and also airway inflammation diseases, suggesting that LOX inhibition may have beneficial effects as a potential target to limit the severity of related symptoms of these diseases and therefore these inhibitors could be considered as an agent for decreasing the enzyme activity in association with the disease.
Keywords : Bivalent ions, Chemical Inhibitors, Chromatography, Humane placenta, LOX activity, LOX isolation.

Abstract
Purpose The present study is a case–control analysis of a SNP
(rs28368082) in exon 7 of the SPO11 gene and its possible
association with male infertility in three provinces of Iran.We
also searched for genetic differences among populations.
Methods Using Polymerase Chain Reaction-Restriction Fragment
Length Polymorphism (PCR-RFLP) analysis, we genotyped
113 infertile men and 50 fertile controls. Then, samples
consisting SNP, as determined by PCR-RFLP, were genotyped
by sequencing. The differences in genotype distributions
between cases and fertile controls were examined using
Chi-squared analysis. The genetic difference between individuals
with mutated nucleotide was investigated by phylogenetic
trees. Genetic difference among populations (provinces)
was analyzed through ANOVA test, and homogeneity was
investigated using STRUCTURE and K-means clustering
analysis.
Results According to the statistical analysis, the SNP was
significantly associated with male infertility in all populations
except oligozoospermic cases of the Center region. The phylogenetic
trees showed partial genetic variation among the
individuals, although ANOVA test showed no significant
genetic difference between populations (provinces) for both
azoospermic, and oligozoospermic cases. Eventually, we affirmed
that individuals in the inclusive populations had genetic
difference, but it was not statistically significant for dividing
underlying populations to separate groups, so each population
was homogenous.
Conclusion Our study indicates that the mentioned polymorphism
in SPO11 gene may be linked to the susceptibility
of azoospermia and oligozoospermia male infertility
in three provinces of Iran. Further studies are required
to support obtained results. It finally should be
noted that the possible association between a particular
SNP and a specific disease completely depends on the
underlying population.
Keywords SNP .Male Infertility .SPO11gene .Azoospermia .
Oligozoospermia

Purpose A case-control population study was performed to investigatie the
association of the SNPs rs: (12676) and (rs: 9001), (rs: 200569248) and (rs:
78371839) located on CHDH gene with azoospermiamale infertility in Iranian
population.
Methods Two separated population of Center and North of Iran (In Royan Institute)
were collected. DNA was extracted from each samples through salting-out method.
a 345bp segment of the gene was amplified using PCR. All samples were
genotyped using direct sequencing. Chi-squared test and Odds Ratio were recruited
to check the association of the SNPs with azoospermia, and to investigate the
genetic difference in populations AMOVA was performed.
Result There was no significant genetic difference between our populations, which
meant they were almost genetically similar, so we merged data from both
populations in one single group. We also find a positive association between SNPs
(rs: 12676) and (rs: 9001) and azoospermia infertility but no association was found
in SNPs (rs: 200569248) and (rs: 78371839) with azoospermia male infertility.
Conclusion Positive association probably reveals an important role of this gene in
male infertility. Further studies are needed to support the obtained result. It is
important to know if such an association plays a key role in male infertility so
probably with a Betaine rich diet treatment we can overcome to the CHDH enzyme
defects.

Abstract The majority of cases of ALL demonstrate an abnormal karyotype, either in chromosome number or structural changes. Abnormal chromosome number in childhood acute lymphoblastic leukemia defines distinct biological subgroups with a different response to treatment. The tubes are cultured with three different protocols to save time if one protocol failed. Cultures are then harvested, and cells are fixed and chromosome spreads are prepared. Of 25 patient studied, one patient had psudodiploid karyotype, three patients had tetra-ploid karyotype, four patients had low hypo-diploid karyotype, four patients had high hyper-diploid karyotype, five patients had low hyper-diploid karyotype and eight patients had normal karyotype. Some other factors like, Age, Sex, Consanguinity, Hemoglobin, WBC count, and Type of the leukemia cell also have been evaluated. We found excess number of patients having hypodiploid karyotype but still response to treatment protocols were satisfactory. By comparison between 4 different cultures methods, we find direct method to be more efficient for ploidy analysis. Keywords ALL, AML, Karyotype, Ploidy, Leukemia

Abstract
Background and purpose: Nitric oxide (NO) is produced by nitric oxide synthase (NOS) from
L-arginine and has an important role in a variety of physiological and pathological processes in biological
systems. In this study, kinetic activities of isolated NOS were evaluated from sheep kidney.
Materials and methods: In this reasearch homogenization, ammonium sulfate precipitation and
column chromatography on DEAE-32 Cellulose and 2', 5'-ADP-agarose of 500 grams of sheep kidney
were used to purify isolated nitric oxide. In all steps of purification process the amount of protein and its
activity was assayed using Bradford and Griess reactions.
Results: The molecular weight of sheep kidney on SDS-PAGE electrophoresis was 54 KD.
Specific activity was 0.6 units/mg protein and recovery of purification was 0.9% (relative purity was
20 times more). Optimum temperature and pH were 30°C and 7.4 and incubation of purified enzyme at
thermal interval of 10 to 30°C for 15 minute showed a stable enzyme activity. At 75 μM concentration of
L-arginine substrate, the highest level of NOS activity was seen. The Vmax and Km of enzyme were
250 nmol/min/mg protein and 5.32 M, respectively.
Conclusion: NOS enzyme from sheep kidney was purified with relative purity of 20 times and
their optimum temperature, pH, suitable substrate concentration, Vmax and Km were 30°C, 7.4, 75 μM,
250 nmol/min/mg proteins and 5.32 μmol, respectively.
Keywords: Nitric oxide synthase (NOS), kinetic activity, sheep, kidney

Abstract
Purpose The present study is a case–control analysis of a SNP
(rs28368082) in exon 7 of the SPO11 gene and its possible
association with male infertility in three provinces of Iran.We
also searched for genetic differences among populations.
Methods Using Polymerase Chain Reaction-Restriction Fragment
Length Polymorphism (PCR-RFLP) analysis, we genotyped
113 infertile men and 50 fertile controls. Then, samples
consisting SNP, as determined by PCR-RFLP, were genotyped
by sequencing. The differences in genotype distributions
between cases and fertile controls were examined using
Chi-squared analysis. The genetic difference between individuals
with mutated nucleotide was investigated by phylogenetic
trees. Genetic difference among populations (provinces)
was analyzed through ANOVA test, and homogeneity was
investigated using STRUCTURE and K-means clustering
analysis.
Results According to the statistical analysis, the SNP was
significantly associated with male infertility in all populations
except oligozoospermic cases of the Center region. The phylogenetic
trees showed partial genetic variation among the
individuals, although ANOVA test showed no significant
genetic difference between populations (provinces) for both
azoospermic, and oligozoospermic cases. Eventually, we affirmed
that individuals in the inclusive populations had genetic
difference, but it was not statistically significant for dividing
underlying populations to separate groups, so each population
was homogenous.
Conclusion Our study indicates that the mentioned polymorphism
in SPO11 gene may be linked to the susceptibility
of azoospermia and oligozoospermia male infertility
in three provinces of Iran. Further studies are required
to support obtained results. It finally should be
noted that the possible association between a particular
SNP and a specific disease completely depends on the
underlying population.
Keywords SNP .Male Infertility .SPO11gene .Azoospermia .
Oligozoospermia

Abstract
BACKGROUND:
It has been hypothesized that microdeletions of Yq may account for a significant proportion of men with infertility. Three
nonoverlapping regions, referred to as “azoospermia factors” (AZFa, b, c from proximal to distal Yq) have been defined as
spermatogenesis loci and deletions in these regions have been shown to be pathogenically involved in male infertility associated with
azoospermia or severe oligospermia.
AIMS:
Evaluation the frequency of Y chromosome microdeletions in Iranian population.
MATERIALS AND METHODS:
Fifty infertile men were selected. Semen analysis was done and on the basis of the mean sperm count, all patients were categorized
into azoospermia and oligozoospermia, groups. Blood samples were obtained for DNA extraction and chromosomal analysis. GenomicDNA was extracted from blood lymphocytes and amplified by sequence tagged sites-polymerase chain reaction (STS-PCR) method to
determine the presence of microdeletions in AZF locus. A total of 34 STS primers including two controls were selected to identify
microdeletions of Y chromosome on each subject.
RESULTS AND CONCLUSION:
26/50 cases (52%) showed deletion of at least one of the STS Marker. Totally 41 microdeletions was observed. A total of 17 cases (34%)
had deletion in one STS. Four oligospermia cases (8%) had deletion in 2 STS site. Three azoospermia cases (6%) had again deletion in
2 STS site, but in different STSs. One case had three deletions in three STS site and finally one individual had seven deletions in AZF
locus. The overall frequency of Y chromosome microdeletions observed in the present study was found to be 26/50 (52%). Comparison
of our data with the result of other investigators world wide shows that the incidence of Yq microdeletions in Iranian population is
much higher than international frequency. Our data agree with other studies regarding microdeletions of AZFc, but for microdeletions
of AZFa (14.6%) our results is much higher and differ significantly with many studies.
Keywords: Azoospermia, infertility, oligospermia, Y-chromosome microdeletions

Objective: The colony from inner cell mass of embryo has potential to proliferation unlimitedly in vitro and remain undifferentiated. These colonies can maintain their normal karyotype in eupleuidy in a cell population. In order to prepare the embryonic stem (ES) cells, we cultured the blastocyst and evaluated the morphology of inner cell mass during the outgrowth, disaggregation, expantion and production of the cell colonies. The karyptype of these cells was studied in the early passage.
Methods: For this purpose, 20 mitomycin C-treated intact blastocyst, taken from 15-day rat embryos, were transferred to a layer of embryonic fibroblasts. The outgrowth of inner cell mass was mechanically disaggregated and the cells were passaged every 2-3 days in DMEM 1000 IU/ml containing LIF and 20% FBS. The cells were sub-cultured for up to nine passages. The colony of the cells during the 7th and 9th passages were processed for karyotyping by Giemsa staining.
Results: The results showed that in the expantion stage the coloney appeared as a flat monolayer mass with strike boundaries and nondistinguished cytoplasm with a few nuclei. In coloney formation stage, the undifferentiated colonies of cell change their morphology to a round multilayer colony with clearly defined boundaries. Chromosomal status showed normal karyotype in 86% of cells at the 7th passage and in 67% of cells at 9th passage. The anupleuidy of the chromosomes was mostly observed as monosomy and metacentric.
Conclusion: The overall findings from this study demonstrated that the ES undifferentiated colonies could be distinguish by morphological criteria, passage acceptability and division diplication rate. Furthermore, detection of ES cell colonieo during the expantion stage is the first signs of success in isolation of ES cell coloneies.

Keyword: INNER CELL MASS, EMBRYONIC STEM CELL, KARYOTYPE, MOUSE

ABSTRACT
Background and Objective: Pregnancy termination and recurrent abortion are one of the
common complications during pregnancy and in patients with a bad obstetric history.
Materials and Methods: In this study, a total of 154 individuals including 75 couples and
four single women from different communities and with various incomes were investigated for
chromosomal abnormalities using blood culture and chromosomal banding technique.
Results: Chromosomal analysis of these patients revealed three abnormal karyotypes
(3.8%) in three women and two abnormal karyotypes in conceptions. Two of these couples had
consanguineous marriage and the remaining women included one isochromosome for X [46, x,I
(xq)], two translocations [45, xx, t (15:21)] and [46, xx, t (7:14)], one trisomy ‘21’ (47, xx, +21), and a
ring chromosome (46, xx, r(X). In addition, 27 conceptions had been reported for these five couples.
These included 23 abortions with 18 of them within first trimester (78.26%) and four of them had
abortions within second trimester (21.74%), one had a normal child, three had abnormal children,
and one with stillbirth.
Conclusion: It was found out that abnormal karyotype is present in 3.8% of patients with a bad
obstetric history. There was also a close relationship between number of deliveries and abortions
and this relation was statistically significant (p<0.01). In addition, consanguinity was also related
with number of abnormal children (p<0.05). There was also a significant relationship between
consanguinity and first trimester abortions (p<0.05). Therefore, in couples with more than three
abortions, especially within first trimester, chromosomal evaluation can have a diagnostic value.
Key words: Obstetric history, Karyotype, Chromosomal abmormalities, Abortion

ABSTRACT
The relation of ABO blood groups to disease is well established. In 1943, Levine had identified ABO incompatibility
as a cause of early abortions and stillbirths. From this time onwards numerous workers produced
data suggesting, mainly on the grounds of a deficiency of A children, and an excess of abortions, in the
families of O women married to A men, that the A fetuses produced by such mating were especially liable to
be aborted. Seventy-nine couples from Pune city (India), suffering from repeated abortion have been investigated
for the ABO blood groups system to find out the frequency of ABO blood group phenotypes and ABO
incompatibility as a cause of abortion. In husband group maximum number of individuals had blood group A.
In wife group, blood group B showed the highest number of individuals. In mixed group 154 patients have been
tested and out of these, again blood group A, shows the highest number of individuals. Blood group “A”
and “AB” was significantly higher in individual and mixed groups as compared with normal groups in this study.
ABO blood group of husband/wife mating was also determined, the analysis of husband/wife joint “ABO” blood
group distribution in these couples, shows an excess of joint “A/B” blood groups in couples as compared
with expected proportions assuming random mating. This study came to a conclusion that there is a clear increase
in number of individuals for blood group A and AB in-patients with repeated abortion, and this factor may
need to pay more attention in future investigations. It is possible that incompatibility of the antigens present
in red blood cell membrane of husband/wife may play some role in abortion.
Key Words: Blood group, Abortion, Incompatibility