[ effect of oxamate on LDH-C4 activity of sperm in rats]

LDH-C4 is one of the lactate dehydrogenase isoenzymes found in mature testis and spermatozoa of many species. The Physiological function of these isoenzymes indicates its role in creating energy for sperm motility and survival. In this research, the effects of oxamate as specific inhibitors of the LDH-C4 of rat were studied in vivo. A total of 20 adult rats were divided into four equal groups. One group as the control group received saline only, and different amounts of oxamate were injected into other three groups [600, 300, 150mg/kg] daily for 45 days intraperitoneally. The rats were then killed with chloroform and the caudal part of epididym was separated. By making several cuts in caudal part of the epididymis, the sperms were isolated and put in T6 medium+5mg/ml[-1] BSA. Later, the sperms were incubated under 37§C and 5% CO2 for one hour. LDH-C4 enzyme was extracted using the Erwin Goldberg and the protein amounts were measured by Lowry's method. Relative purification was done in two stages including ammonium sulfate precipitation and column chromatography by DEAE-Sephadex-A50. All stages of extraction, the amount of total protein, LDH enzyme activity in the oxamate-exposed groups, and specified recipient, were then compared with the control group. In this study, the total enzyme LDH-C4 activity in the control group was 11.8 +/- 0.3 and the oxamate recipient groups [150, 300 and 600mg/kg] were 8.3 +/- 0.3, 6.9 +/- 0.2 and 3.2 +/- 0.1 IU, respectively. The concentration of 600mg/kg oxamate was inhibited about 63% enzyme activity compared to control group. This study showed that the oxamate may reduce in vivo enzyme activity through LDH-C4 with increasing concentration and this effect is proportional. Therefore, with the effect of the competitive inhibitors of oxamate on LDH-C4, this substrate can be used as a contraceptive for males

Mini-Exon genotyping of leishmania species in khuzestan province, southwest Iran

Leishmaniasis is a protozoan disease cause by Leishmania genus. Anthroponotic and zoonotic cutaneous leishmaniasis are endemic in Iran. The aim of this study was to identify the causative agent of cutaneous leishmaniasis by mini-exon gene in five regions of Khuzestan Province, southwest of Iran. From 2007 to 2008 in this cross-sectional study, cutaneous samples were collected from patients referred to Health Centers and Hospitals of the Khuzestan Province for cutaneous leishmaniasis diagnosis and cultured in Novy-MacNeal-Nicolle [NNN] and RPMI 1640. The propagated promastigotes were harvested and Leishmania species of cutaneous leishmaniasis were identified by RFLP and DNA sequencing of the PCR generated fragments. L. major and L. tropica were the causative agents of cutaneous leishmaniasis by predominantly of L. major species. The alignment of the mini-exon sequencing isolates with reported sequencing of L. major and L. tropica revealed 92%-99% identity. Our study showed that mini-exon PCR-RFLP was useful method to identify the causative species of cutaneous leishmaniasis

Induction of apoptosis by miltefosine in Iranian strain of Leishmania infantum promastigotes

Miltefosine is a new drug of choice for the treatment of visceral leishmaniasis. Numerous experimental studies have shown miltefosine is effective on Leishmania donovani, however, effectiveness of miltefosine in treatment of L infantum is not fully understood. The aims of the present study were to evaluate cytotoxic effects of miltefosine on Iranian strain of L infantum, and to determine its 50% inhibitory concentration [IC50] as well as lethal dose. Anti-L. infantum activity of miltefosine was studied by treatment of cultured promastigotes with various concentration of miltefosine. MTT assay was used to determine L infantum viability and the results were expressed as IC50. Annexin-V FLUOS staining was performed to study apoptotic properties of this drug by using FACS flow cytometry. Miltefosine led to dose-dependent death of L. infantum with features compatible with apoptosis including cell shrinkage, DNA laddering, and externalization of phosphatidylserine with preservation of integrity of plasma membrane. The 100% effect was achieved at 22 micro M and IC50 after 48 hours of incubation was 7 micro M. Miltefosine exerts cytotoxic effect on Iranian strain of L. infantum via an apoptosis-related mechanism

[Study the effect of oxamate on rat sperm motility in vivo]

LDH- C4 is an isoenzyme of lactate dehydrogenase that is found in mature testes and spermatozoa of species with internal fertilization. Its physiological function appears related to metabolic processes that provide energy for motility and survival of spermatozoa. Oxamate is a new selective competitive inhibitor of sperm LDH- C4 with pyruvate as substrate. In the present experimental study on male rat, the effectiveness of oxamate was evaluated as a novel approach to the development of a male contraceptive. In this study, 20 adult rats were divided into 4 groups, the first used as control and the remaining three used as experimental groups. Experimental groups received different concentration, of oxamate [150, 300, 600 mg/kg/,ip] for 45 days. Control animals received normal saline solution. The sperms from the cauda division of epididymidis were collected by placing minced cauda in culture medium [T6] for one hour at 37°C in a 5% CO[2] atmosphere. Sperm motility was evaluated utilizing Makler chamber and compared with the control group. Statistical analysis was performed by the student t- test and one - way ANOVA. Progressive sperm motility in control and treated their groups were%60.3 +/- 2.8],%50 +/- 2.4],%41.5 +/- 1.9],%19 +/- 2.2] respectively. We conclude that oxamate in vivo can reduce sperm motility significantly and this reduction was concentration-dependent. The results of this work show that sperm motility can be reduced by concentration- dependent effect of oxamate under in vivo conditions

Determination of antibodies [IgG, IgM] against toxoplasma gondii in patients with cancer

The aim of this study was determination of antibodies [IgG, IgM] against Toxoplasma in malignant patients in order to refer the patients on time to the physician for treatment. This study was carried out on 252 malignant patients and 252 healthy normal subjects [as control] obtained from Shafa Hospital and Medical Diagnostic Laboratory [Iran-Zamin], in Ahwaz city. Patient's information was recorded in a questionnaire before sampling. Serum samples of patients were examined for IgG and IgM antibodies by ELISA technique using Trinity kits. The results of this study revealed the presence of Toxoplasma antibodies in 114 [45.2%] cases of patients who were positive for Toxoplasma IgG antibodies, and 26 [10.3%] cases were confirmed to be positive for Toxoplasma IgM antibodies and also 17 [6.7%] of cases had both IgG and IgM antibodies against Toxoplasma gondii. In control group 92 [36.5%] cases and 15 [6%] cases revealed seropositive for IgG and IgM antibodies, respectively. There were no significant differences between sex, close contact with cat, living region, chemotherapy, and seropositivity rate of toxoplasmosis in patients. Comparing the age groups, the highest seropositive rate showed in the age of 51 years or higher, and their rates had tendency to increase with age in both groups. No seropositivity significant relationship was found between patients and control group. According to the prevalence of positive cases in these patients, it is necessary to examine the patients for toxoplasmosis before, during and after chemotherapy