Monarch-1 activation in murine macrophage cell line [j774 a.1] infected with Iranian strain of leishmania major

Leishmania major is an intracellular parasite transmitted through the bite of the female phlebotomine sand flies. Leishmania major is able to escape the host immune defense and survive within macrophages. Modulation of the NF-kappaB [Nuclear Factor-Kappa B] activation and suppression of the pro-inflammatory cytokines by L. major are the main evasion mechanisms that remain to be explored. This study aims to examine the expression level of the Monarch-1 in L. major-infected macrophages, as a negative regulator of the NF-kappaB activation. Murine macrophage cell line [J774 A.1] was infected by metacyclic form of Leishmania promastigotes at macrophage/parasite ratio of 1:10. After harvesting infected cells at different times, total RNA was extracted and converted to cDNA. Semi-quantitative RT-PCR was performed for Monarch-1 by specific primers. Hypoxanthine Phospho-Ribosyl Transferase [HPRT] was used as an internal control to adjust the amount of mRNA in each sample. Semiquantitive analysis of Monarch-1 mRNA expression level showed a significant expression increase within 6 to 30 hours after L. major infection of macrophages when compared to the control macrophages. Monarch-1 expression level reveals a significant increase in the early phase of macrophage infection with L. major, which in turn may suppress IL-12 production in Leishmania infected macrophages and deeply influence the relationship between host and parasite.

Molecular identification of leishmania species using samples obtained from negative stained smears

Cutaneous Leishmaniasis [CL] is a parasitic skin disease. Diagnosis primarily is based on clinical signs and microscopic observation of parasite on direct stained smears or tissue sections. Sensitivity of direct smear is not as high as molecular methods. The aim of this study was to identify and characterize Leishmania species among the negative direct smears obtained from skin ulcers suspected to CL by PCR method. Among 81 patients with suspicious skin lesions to CL referred to the Parasitology lab, negative Giemsa stained smears were collected. DNA extraction performed by scraping stained smears, then PCR was performed. Among the DNA extracted from smears, L. tropica was isolated from 9 [11.1%] of the smears and L.major was not isolated from any samples. Direct microscopy on stained smears for diagnosis of leishmaniasis is not enough accurate. PCR is recommended for clinically suspected lesions with negative result of direct smear.

Mycological microscopic and culture examination of 400 bronchoalveolar lavage [BAL] samples

The frequency of invasive opportunistic mycoses has increased significantly over the past decades especially in immunocompromised patients. Invasive aspergillosis [IA] has become a major cause of morbidity and mortality among these patients. As bronchoalveolar lavage [BAL] fluid samples are generally useful specimens in the diagnosis of invasive pulmonary aspergillosis [IPA], this study was designed to evaluate the incidence of fungal elements in at-risk patients by direct microscopy and culture of BAL samples. In a 16-month period, 400 BAL samples were obtained from several groups of different patients with pulmonary and respiratory disorders and examined by using both direct microscopy and culture. Of the 400 samples, 16 [4%] were positive direct examination with branching septate hyphae and 46 [11.5%] were positive culture: 25 [54%] Aspergillus flavus, 6 [13%] A. fumigatus, 5 [10.9%] A. niger, 1 [2.2%] A. terreus, 3 [6.5%] Penicillium spp. and 6 [13%] mixed A. flavus/A. niger. A. flavus was the most common cause of Aspergillus infection or colonization. Bone marrow transplant [BMT] recipients were the most susceptible group to fungal infection and/or colonization. Among Aspergillus species, A. flavus was the most common isolate in both infections and colonization in Iran. More studies are needed to clarify the epidemiological aspect of aspergillosis in Iran

Whatman paper [FTA Cards] for storing and transferring Leishmania DNA for PCR examination

Diagnosis of cutaneous leishmaniasis [CL] is often made based on clinical manifestation. Correct diagnosis and identification of the parasite are crucial for choosing the effective treatment and for epidemiological studies. On the other hand, determination of Leishmania species is necessary for designing appropriate control programs. Diagnosis by PCR is becoming a [gold standard]. For PCR preparation, storage and shipments of specimens are necessary. In this study, Whatman filter paper [FTA Card] was used to store and transfer samples for Leishmania identification using PCR. Among the patients who had CL lesion and referred to Parasitology Laboratory of Emam Reza Hospital, Mashhad, Iran, 44 consented cases with positive results in their direct smear were selected. An informed consent form and a questionnaire were completed and three different types of samples [direct smear, NNN culture, and spot on FTA card] were collected. DNA extraction and PCR were carried out on three different samples from each patient. PCR results using Whatman paper samples revealed a significant difference [P<0.0001] compared to the culture method but no significant difference was seen between PCR results using samples stored on Whatman paper and direct smears. The use of FTA cards is simple, rapid, and cost-effective, and can be readily employed for large-scale population screening, especially for regions where the specimens are to be transported from distant places to the laboratory

Flow cytometry susceptibility testing for conventional antifungal drugs and comparison with the NCCLS broth macrodilution test

During the last decade, the incidence of fungal infection has been increased in many countries. Because of the advent of resistant to antifungal agents, determination of an efficient strategic plan for treatment of fungal disease is an important issue in clinical mycology. Many methods have been introduced and developed for determination of invitro susceptibility tests. During the recent years, flow cytometry has developed to solving the problem and many papers have documented the usefulness of this technique. As the first step, the invitro susceptibility of standard PTCC [Persian Type of Culture Collection] strain and some clinical isolates of Candida consisting of Candida albicans, C. dubliniensis, C. glabrata, C. kefyer and C. parapsilosis were evaluated by macrodilution broth method according to NCCLS [National Committee for Clinical Laboratory Standards] guidelines and flow cytometry susceptibility test. The data indicated that macro dilution broth methods and flow cytometry have the same results in determination of MIC [Minimum Inhibitory Concentration] for amphotericin B, clotrimazole, fluconazole, ketoconazole and miconazole in C. albicans PTCC 5027 as well as clinical Candida isolates, such as C. albicans, C. dubliniensis, C. glabrata C. kefyr, and C. parapsilosis. Comparing the results obtained by macrodilution broth and flow cytometry methods revealed that flow cytometry was faster. It is suggested that flow cytometry susceptibility test can be used as a powerful tool for determination of MIC and administration of the best antifungal drug in treatment of patients with Candida infections

[ case report of alveolar echinococcosis]

Echinococcus multilocularis is the cause of alveolar hydatic cysts in intermediate host, found in the liver and lungs. Alveolar hydatid diseas is the most lethal of helminthic diseases, with radical surgery still being the only curative therapy. The aim of this study was to report a splenic alveolar hydatic cyst. The patient was a 62 years old man, from a rural region of Ghochan, Khorasan Province, Iran, who was referred to surgeon, complaining left upper quadran abdominal pain. In CT scan 2 cysts were seen in the right lobe of liver and a septal cyst in spleen. After operation, evacuating and unroofing of the liver cysts, splenectomy was performed. Diagnosis was based on pathologic and histologic examinations. In macroscopic examination spleen was massive and cystic, composed of multiple irregular cysts with infiltrative appearance. The inner part of cysts contained brown necrotic and gelatinous material, at microscopic examination there was a thin acellular laminated layer but the germinal layer and protoscolex were not seen. After a week the patient recovered and left the hospital with good general condition. In conclusion, the presence of this case showed that life cycle of echinococcus multilocularis can be completed in this province. Medical managers and practitioners must notice this disease as a differential diagnosis for malignant of liver and spleen, as well as for prevention and control of the disease itself

[Psoriasis and infestation with Malassezia]

Psoriasis is a chronic proliferative condition of the skin. Its accurate pathogenesis has not been known yet but interactions between genes and environmental factors have been implicated in its initiation. Although several reports have associated this lipophilic yeast with development of skin lesions in Psoriasis, the definite role of this lipophilic yeast in psoriasis is still undetermined. Our aim was to investigate the correlation between Malassezia and psoriasis. In this six-month's case-control study, a sample was obtained from cutaneous lesions of 50 psoriatic patients and 50 healthy volunteers as control group for evaluation of Malassezia infestation. Data were collected by use of questionnaires and analysed by SPSS software and using statistical tests of X[2] and Mann-whitney. There is no difference between Malassezia infestation in scalp lesions of psoriatic and healthy persons [P=0.86]. Malassezia infestation in psoriatic patients with scalp involvement was more than those without scalp involvement, but this relationship was not statistically significiant [P=0.069]. There is an inverse significant relationship between scalp infestation with Malassezia and chronicity of psoriasis [P=0.04]. Infestation with Malassezia in body skin of psoriatic patients was less than normal persons [P< 0.000]. It seems that Malassezia has an initiating role in inducing immune mechanism involved in pathogenesis of scalp psoriasis by Malassezia, but with chronicity and formation of dry and hyperkeratotic plaques, the environment will be inappropriate for Malassezia, so Malassezia infestation decreases with chronicity of disease

[Another look at the cases of Mucormycosis at Emam Reza hospital]

Mucormycosis or Zygomycosis is a rare opportunistic and fulminated fungal infection with high mortality. It occurs most often in diabetic and immuno-compromised patients, including those with hematological malignancy. There are several forms of disease that rhinocerebral is the most common form. Early diagnosis plays an important role in prognosis of the disease. Since, previous papers were mainly focused on pathologic aspects and case reports, we decided to review the cases on the basis of etiologic agents, predisposing factors, Clinical signs on admission and the role of early diagnosis in golden time on prognosis. This is a retrospective and cross sectional study of 18 cases of approved Mucormycosis that diagnosed in Imam Reza hospital during 12 years during 1312-1384. In addition to those, four recent cases studied prospectively. The study population was 15 men and 7 women. The cases were from 1 to 80 years old. Diabetes mellitus was the most common Predisposing factor [54.4%]. Most of the cases were rhinocerebral form. Mortality rate was more than 75%. Prognosis was good in less than 25% of the cases who received early diagnosis and treatment. Early diagnosis and correct Treatment [surgical and medical intervention] are critical because they could improve the prognosis for survival

[ study on the vectors of the cutaneous leishmaniasis in the northern part of Mashhad, Iran]

Phlebotomine sand flies are the vectors of different forms of leishmaniasis in Iran. Study on their ecology is essential for suggestion of leishmaniasis control programs. Following annually new case reports of cutaneous leishmaniasis, at Hemat Abad an investigation on the fauna and monthly activating phlebotomines was carried out during Mar 2003. Nov. 2003 in this region. Phlebotomine sand-flies were collected every 10 days from outdoors and indoors with sticky traps, during Mar. 2003-Nov. 2003. In this study 521 sand-flies were collected from outdoors and indoors [2 species of phlebotomus genus and 2 species of sergentomyia genus], as follows, P. sergenti [74.7%], P. papatasi [1Ll %], S. sintoni [13.3%], S. sombarica [0.9%]. The sand flies active season begins from April till October in this region. P. sergenti was the predominant phlebotomines species in our investigation. Based on the results, P. sergenti is the probable vector of Cutaneous leishmaniasis in this region and P. Papatasi can transmit disease from rodents to humans

Treatment of cutaneous leishmaniasis in murine model by alcoholic extract of Berberis vulgaris

In order to evaluate the effect of Berberis vulgaris extract on the experimental ulcers of cutaneous leishmaniasis [CL] on Balb/c mice, a study was undertaken over a 12 months period. Forty Balb/c mice were divided into 2 main groups A and B. Each main group in turn was divided into 5 sub groups of 4 mice and each sub group were inoculated subcutaneously by 0.1ml liquid phase culture containing promastigotes of Leishmania major. After 2-3 weeks, nodules and ulcers appeared on 37 of 40 inoculated mice. Ethanol extract of the stem and leaves as well as roots of Berberis vulgaris in different concentrations, were used topically on CL lesions of 4 sub groups A and B, respectively. Ethanol alone was used on the lesions of control mice. The surface area of lesions were measured before and 1-2 weeks after treatment. Direct Geimsa stained smear prepared 20 days after treatment. The results showed that after 2 weeks, a statistically significant decrease of ulcer size of treated mice observed, while in the control group the lesion growth continued. The examinations showed that using higher concentration of the extract caused more decrease in surface area of CL lesions on day 15 and negative direct smear on day 20. Alcoholic extract of B.vulgaris root was more effective than leaves and stem extract. Alcoholic extract of B vulgaris might be further used in animal model