Cell Journal (Yakhteh)
Volume:2 Issue: 4, 2000

Lishmania is a protozoan parasite that cause visceral and cutaneous disease in man. This parasite shows two forms in its life cycle, amastigote in reticulo-endothelial cell of mammalian host and, promastigote in culture media and sand fly vector. Like other kinetoplastide, leishmania has an extra chromosomal DNA with unique topological and characteristic that called kDNA. The organism is responsible of different clinical manifestation in man, therefore, identification of different species and strains was always demanded. Identification and characterization of leishmania depend on geographical disteribution, clinical manifestation, monoclonal antibody, serological methods, isoenzyme pattern and others. In last decade molecular methods was developed for these aims. DNA is stable through its life cycle and kDNA organized in two different molecules, minicircles has 0.5-2.9 kb with heterogeneity in sequences and variable in number, and maxicircles with 20-50 kb size that codes for enzymes and cofactors of mitochonderial genes. KDNA minicircle of leishmania major was extracted and DNA library was constructed by BamHI enzyme in pBluescript plasmid. After transformation, recombinant plasmid was screened by a-complementation test on X-gal and IPTG - treated agar plates. Two clone of leishmania major kDNA in BamHI site of pBluescript plasmid were identifed. Ribiperobe syntesized by run off synthesis method pBluescript plasmid. To define probe specificity. They were examined with inserted DNA and total kDNA through dot blot and southern blot hybrydization by IRI-m2 RNA probe. IRI-m2 was hybridized with L. major with high specificity and lesser specifity with L. infantum. This sequence is suitable for PCR primer design and use for differentional of L. major and L. donovani by L. tropica.

Cd59 is a complement control protein (CCP) wich exhibits its inhibitory functions by prevention of complement attack complex (MAX) formation. The aim of this normozoospermia, olig-astheno and azoospermia. The semen of 57 individual were grouped to normozoospermia (8), asthenozoospermia (21), oligo-astenozoospermia (16), and azoospermia (12) according to WHO ceriteria. After seperation of seminal plasma, the CD59 conccentration was measured by the sandwich ELAISA method, using two monoclonal antibody againts CD59. One as capture and the other for detection of CD59 protein. The mean concentration of CD59 within the Normozoospermia, asthenzoospermia, oligo-asthenozoospermia, and azoomospermia was: 33.53 ±4.26, 28.14±6.59, 20.32±7.43, reverse linear correlation between sperm density and CD59 concentration (r=-0.58, P=0.000). This is the first report, which shows differences in CD59 concentration between normal and abnormal semen samples. In this study we obtained a significant difference in the CD59 mean concentration of the normal group with both azoospermia and oligo-asthenozoospermia groups. As well as a negative correlation between CD59 concentration and sperm density and a positive correlation between CD59 concentration and sperm viability.

To evaluate the quick Cryopreservation technique on parameters of motility, morphology, and vitality of spermatozoa from normozoospermic ejaculates. In this prospective study, a total of 37 normal ejaculates were evaluated according to WHO criteria, then each sample was divided into two aliquets and sample was cryopreserved with quick freezing method. Sperm parameters of count, motility, morphology and vitality were measured following thawing. In addition, the correlation between Motility survival rate and sperm parameters was studied following freezing. The results showed that the rates of count, normal morphology, progressive motility, and percentage of live spermatozoa were significantly reduced following thawing (P<0.001). The mean of sperm progressive survival rate was 35.62±21.72% which was in direct relation with parameters of progressive motility, morphology, and vitality before the freezing (P<0.05). This was not correlated with non- progressive motility and sperm count (P>0.05). In spite of reduction in sperm parameters following freezing, a vast number of spermatozoa preserve their vitality. This condition is important for infertility treatment programs. In addition, normal parameters of progressive motility, vitality, and morphology play a vital role in molility survival rate of spermatozoa.

The osteopetrosis is a common disease in man and animals which is characterized by calcium accumulation in bones. The purpose of this study is to eveluate the osteoblast ultrastructure in osteoptrotic Op/Op mouse. In this study 5 Op/Op (experimental) and 5 control mouse were killed by overdose of choloroform. Their femur bones were taken, decalcified by EDTA, fiexed in glutaraldehyde and osmium tetroxide, embeded in EPON, sectioned by diamond knife, stained by lead citrate and uranil acetate and photographed by transmission electron microscope (Hitachi 800, Japan). The ultrastructural observation of osteoblast cells revealed a decrease in the mitochonderia and endoplasmic reticulum in of Op/Op mouse, but the cytoplasmic vacuoles and free ribosomes were high. The nuclear envelope in Op/Op group was irregular and higher chromatin density. Also, the osteoid in Op/Op group was thickened. The activity of osteoblast was decreased due to the inactivity of osteoclast as a result of lack macrophage colony stimulating factor.

Retinoic acid (RA), one of the synthetic vitamin A derivative which has been frequently used as a drug to treat face acne, is know to have specific side effects on the testis, liver, spleen and etc. Therefore, we have tried to find a compound which would decrease its side effects. Eugenol is a compound extracted from Clove tree. It has anti-anaphylactic, anti-inflammation, antiseptic properties and inhibits lipid peroxidation by acting as a chain breaking antioxidant. In trying to find an antagonistic angent for RA, we have used eugenol. In this research, we have chosen 6 weeks old male mice of NMRI strain weighting 26?2 grams. The mice were divided into five experimental groups. The first group were fed only with normal diet (as control)1, but the other four groups were fed with normal diet plus olive oil (as Solvent) 2, (Retionic acid)3, (Eugenol) 4 and (Eugenol+Retinoic acid) 5. We administered 100 mg/kg of eugenol and 30 mg/kg of retionic acid for eight days. Following dissection of mice and collection blood various parameters including cholesterol. Triglyceride, LDH and ALP were analyzed. Results obtained from these experiment indicated that RA increased the amount of cholesterol and triglyceride by 23.3% (P<0.001) and 13.4% (P<0.01) respectively compared with the control group. Similarly, the amount of LDH and ALP activity enzymes occuring in blood plasma increased by 25.8% & 42.4% (P<0.001) respectively compared with solvent group (Control). Dietary uptake of eugenol has decreased the amount of cholesterol and triglyceride by 15.9% (P<0.001) and 12.3% (P<0.01) respectively. Mice were adminstered with dietary eugenol have demonestered a marked decreased in LDH and ALP activities by 13.7% (P<0.01) and 16.4% (P<0.05) respectively. It was obsereved that the cytotoxicity of retionic acid had been decreased markedly by eugenol. Therefore, the antioxidant role of eugenol can decrease the side effects of retionic acid. It can be concluded that eugenol can be used to alleviate side effect of RA.

The possible effect of postnatal chemically induced hypothyroidism on NADPH- diaphorase containing neurons in different cortical areas of rat neonates. After parturition, dams divided in two groups. The control group receiving drinking water, and experimental group receiving 0.1% PTU in their drinking water. In different postnatal days (P4, P7, P10, P15, P23, P30), the pups of both groups were examined for weight and growth indices. For NADPH-d histochemical analysis, the pups of each group were anaesthetized and perfused via heart with fixative solution, then their brains were removed, weighted and sectioned in the coronal plane (80? m) with a vibratome. The sections were processed for NADPH-d histochemical staining. From P7, the mean body and brain weights of treated pups were significantly lower than their matched controls. The number and distribution pattern of ND+ neuron changes in different areas. In the first postnatal days, neurons are usually located in deeper layers (V,VI) and gradually increase in superficial layers (II, III). With exeption to P15, in PTU treated group a significantly increase in ND + neurons population is observed in cingulate, Forelimb/Hindlimb and parietal cortical ereas. PTU-treated pups show a shorter dendritic length and less cortical vascularization which is more severe in P23 and P30. We can conclude that the changes in the pattern of ND + neurons is correlated with developmental processes and cereblar lamination. Rapid and significant increase in ND+ nenronal population in hypothyroid group shows that No-containing neurons are highly sensitive to hypothyroidism and possibly play an important role in induction of hypothyroidism effects on cerebral development and function.

Epilepsy is one of the most common neurological disorders. In spite of the fact that a great development has been made regarding therapy against epilepsy, little is known, however, about the precise mechanism of the epilepsy. Pentylenetetrazol (PTZ) is one of the most often used epileptogenic agents. Hence the main purpose of the present work was to determine the effect of PTZ as a convulsant drug on the bioelectrical activity and neuronal excitability of D5 neurone in Helix aspersa. Experiments were carried out on D5 neuronal soma membrane located in the left parietal ganglion of snail, Helix aspersa. The properties of the action potentials were analyzed using two electrodes current clamp technique. The results showed that the injection of 1 nA depolarizing current in the presence of PTZ caused an increase in the frequency and the duration of action potentials by about 11.7% and 14% respectively. It also led to a 32.3% decrease in the amplitude of afterhyperpolarization and the amplitude of action potential by about 2-3mV. Furthermore when PTZ (25 mM) was applied extracellularly, the resting membrane potential of the D5 cell became hyperpolarized by about 4 mV. These findings suggest that PTZ induces changes in bioelectrical properties which effect on neuronal signaling.

Pentylenetetrazol (PTZ) is one of the most often used epileptogenic agents. Several effects of this epileptogenic drug have been described, but the mechanism of the epileptogenic action of PTZ at the cellular level is still unclear. In this study the effect of PTZ, on the ionic currents of D5 neuronal soma membrane in Helixaspersa, using intracellular recording technique was investigated. Experiments were carried out on D5 neuronal soma membrane located in the left parietal ganglion of the snail, Helix aspersa. The properties of the inward and outward cationic currents induced by PTZ (25 mM) were analyzed using two electrodes voltage clamp technique. The extracellular application of PTZ (25 mM), caused an increase in spontaneous firing activity and with 4 to 7 minutes after adding PTZ, led to a dramatic changes in action potential shape of D5 neuronal cells. Under voltage clamp condition, the peak amplitude of inward current in the presence of PTZ reduced 16.5 % within 7 min. The current-voltage relationship of inward current also shifted by 23.18 % in a hyperpolarizing direction. Furthermore, the peak amplitude of outward currents showed a reduction of about 6.75%. The results of this study show that PTZ by effecting on the properties of bioelectrical activity causes an increase in the excitability of D5 cells. In addition, the effect of PTZ on inward and outward currents accounts for an induction of epileptic activity.