effect of garlic extract on expression of INF gamma and inos genes in macrophages infected with leishmania major

The study was aimed to show the effect of molecular mechanism of Aqueous Garlic Extract [AGE] on expression of IFN gamma and iNOS genes in Leishmania major. Leishmania major promastigotes [MRHO/IR/75/ER] were added to the in-vitro cultured J774 cell line, the cells were incubated fro 72 hours. Various concentrations of garlic extract [9.25, 18.5, 37, 74, 148 mg/ml] were added to the infected cells. MTT assay was applied for cellular proliferation. After 72 hours of incubation, supernatants were collected and total RNA was extracted from the infected cells. The express of IFN gamma and iNOS gene were studied by RT-PCR method. The colorimetric MTT assay after 3 days of incubation showed cytotoxic effect of garlic extract with an IC50 of 37 mg/ml. In addition, IFN gamma and iNOS genes expression by RT-PCR indicated that garlic extract lead to over expression of these genes in 1774 cell line infected with L. major. Garlic extract exerts cytotoxic effect on infected J774 cell line. In addition, the hypthesis that garlic can improve cellular immunity with raising the expression of IFN gamma and of iNOS genes confirmed

[ role of aqueous garlic extract [Allium sativum] on interleukin 10 and 12 in leishmania infected macrophages]

Leishmaniasis is a disease produced by several species of protozoa of the Leishmania genus. As a therapeutic agent, garlic [Allium sativum] is one of the most popular plants in traditional medicinal systems and is increasingly becoming important in herbal alternative therapy. The aim of the present study was to investigate the effects of aqueous garlic extract [AGE] on enhancing T-helper IL-10 and IL-12 cytokines in the culture via macrophages for engulfment and destruction of Leishmania. After proliferation of macrophages in culture and incubation with Leishmania during 72 hours, we added aqueaous garlic extract by several doses 9.25, 18.5, 37, 74 and 148 micro g/ml for 18, 24 and 48 hours. Trypan blue and MTT tests were carried out in order to test the safety of extracts and j744 cells treated with them, as well as cell viability. An enzyme-linked immunosorbant assay [ELISA] was performed on macrophages for interleukin IL-10 and IL-12. The results suggested that garlic extract may provide effective therapy against Leishmania. Our results showed that dose of 37 micro g/ml for 48 hours of garlic extract destructed promastigots by IL-12 secreted from macrophages. Based on the results of this study, IL-12 is crucial for defense against parasitic pathogens. IL-12 administration differentially affected immune response to invading Leishmania parasites

effect of aqueous garlic extract on interleukin-12 and 10 levels in Leishmania major [MRHO/IR/75/ER] infected macrophages

The aim of the present study was to investigate the immunomodulation effects of aqueous garlic extract [AGE] in the cultured macrophages infected by Leishmania major. After J774 macrophages proliferation in RPMI1640 and incubation with Leishmania for 72 hours, AGE was added in doses of 9.25, 18.5, 37, 74 and 148 mg/ml for 18, 24 and 48 hours and cell culture supernatants were harvested. The Leishmania infected J774 cells to assess the cell viability was examined using trypan blue and methylthiazol tetrazolium assay [MTT]. An enzyme-linked immunosorbent assay [ELISA] was performed on cell culture supernatants for measurement of interleukin IL-10 and IL-12. Dose of 37 mg/ml for 48 hours of garlic extract was the most potent dose for activation of amastigotes infected macrophages. In addition, AGE increased the level of IL-12 in Leishmania infected cell lines significantly. AGE treated cell is effective against parasitic pathogens, and AGE induced IL-12 differentially affected the immune response to invading Leishmania parasites

Clinical and imaging findings in 18 patients with eosinophilic pneumonia

Eosinophilic pneumonia is a rare cause of lung disease in adulthood. The relatively non-specific clinical presentations of this disease process make the diagnosis a unique challenge. Herein, we reported on clinical and radiological manifestations of this rare clinical entity in 18 patients. We retrospectively reviewed the medical records of 18 patients with acute eosinophilic pneumonia in Masih Daneshvari Medical Center. Diagnostic criteria were based on clinical, laboratory and imaging findings. The most clinical manifestations were cough and weight loss 15 [0.83] followed by dyspnea 13 [0.72]. The most frequent imaging findings were diffuse opacities [reticulo-nodular or alveolar infiltration] in 11 [0.65], consolidation in six [0.33] and ground-glass opacities in four [0.28] patients. Diffuse rather than peripheral air-space opacities in imaging of a patient presented with dyspnea, cough, sputum, constitutional symptoms and eosinophilia should make us to think about eosinophilic pneumonia as one of differential diagnoses

[Study of differentiation of hippocampal stem cells into rod photoreceptors in rat]

Vision is one of the most important organs in the body. Damage to this organ causes a severe disability in humans. In retinitis pigmentosa, the degeneration of photoreceptors causes blindness. So far, more than 100 mutations have been detected in photoreceptors, which they result in opsin malfunction. The aim of present study is to differentiate the hippocampal stem cells of rat to the rod photoreceptors. Stem cells of the hippocampus were obtained from rat embryos, 18 days of age [E18]. Pregnant female rats were killed and the head of their embryos were separated. Then, the embryos' hippocampus was removed according to Banker's method. Hippocampal cells were dissociated by Fish Bach's method. The cells were cultured in flasks [25cm[2]]. After 3 days, the cells were isolated by trypsin, counted using trypan blue and hemocytometer. Cell suspensions were prepared in two cell concentrations; high [2x10[5]cells/ micro l] and low [2x10[4] cells/ micro l] concentration, then, cultured using DMEM/F12 supplemented with fetal bovine serum 10% [FBS] in six wells plates. Prior to culture of the cells, the first and second row of plates were coated with poly L-lysine and inactivated astrocytes, respectively. Following incubation of the plates at 37°C for 4 days, different concentrations of All Transe Retinoic Acid [ATRA] and 9-CIS RA were added daily for 6 days, and finally immunocytochemistery was carried out using anti-opsin monoclonal antibody. The results of current study showed that the plates, which are respectively treated with ATRA and 9-CIS RA in a concentration of 500nM and 100nM for 6 days had the most differentiated cells. In addition, maximum differenced cells were observed with 100nM of 9-Cis RA. The differentiated cells were detected in wells, which were only coated with inactivated astrocytes in either a high or low concentration of cell suspension. These findings indicated that inactivated astrocytes as a feeder layer and extrinsic factors such as [ATRA] and 9-Cis RA increase differentiation of hippocampal stem cells into rod photoreceptors

Imaging in 100 patients of thoracic hydatid disease including unusual imaging appearances

To evaluate the chest radiography and CT scan characteristics of pulmonary hydatid disease [PHD]. One hundred patients [59 males and 41 females, age ranged from 9 to 80 years] with surgically proven pulmonary hydatid cysts were studied. We reviewed clinical and imaging findings including PA and LAT chest roentgenograms and conventional CT of the chest. Only 82 patients had CT scan in their files, but all had CXR. The radiological features [localization, diameter, architecture, density and other radiological signs and appearances] were determined. On CXR, 124 cysts were determined. In evaluation of 82 available CT scans, a total of 112 cysts were detected. No cysts was detected on 5 CT scans. No discrete cyst was detected on 10 CXRs: 4 patients. only consolidation; and 6 patients, only hydropneumothorax. The most frequent site of involvement was RLL [29.6%]. Fifteen hydatid cysts appeared as solid masses on CT. Fifty-seven cysts were ruptured cysts and 25 patients with ruptured cysts had hemoptysis [43.9%]. Thirty-eight percent of cysts had thin walls and 62% had thick walls. Sixty-four cysts were round in shape [55.7%]. Single cysts were seen in 63 patients while multiple cysts were seen in 37. Median CT density of the cysts was 24 Hounsfeild Units [HU] [-18 to 84]. There were 16 giant cysts [diameter >/= 10 cm] on CT. Mean maximum and minimum dimensions of cysts were 5 cm and 4 cm on CT and 6.8 cm and 5.7 cm on CXR, respectively. On CT and CXR, "water lily sign" was seen in 18 and 22 patients, 'air-fluid level" in 12 and 17 patients, and "crescent sign" in 11 and 5 of patients, respectively. Inverse crescent sign and calcification were not observed on CXRs, but each was reported on 4 CT scans. On CT' 90% of cysts were smooth, 74 cysts were uniloculated and 9 were multiloculated. Nineteen percent of cysts were infected. Other imaging findings included mediastinal shift, atelectasis, infiltration. pericystic lung reaction, chest wall involvement, and rib destruction. CXR is helpful with diagnosis of intact cysts but fails to define entire morphology of complicated cysts. CT imaging recognizes certain details not visible on radiography. In endemic regions like Iran, atypical imaging presentations of complicated pulmonary hydatid disease, such as solid masses, should be considered in differential diagnosis of pulmonary lesions

Effects of retinoic acid and astrocyte as a feeder layer in differentiation of hippocampal stem cell

Adult stem cells are pluripotent cells conventionally isolated from some part of body by different methods. From developmental stand point, murine neural stem cells represent an accessible and important system for studies of basic stem cell property such as self- renewal and multipotency. In this study hippocampal stem cells obtained from embryonic day 18 [E18]. Pregnant female rats were killed, embryos heads separated and then hippocampus isolated by the method of Banker, then their cells dissociated by the methods of Fishbach, and plated in flask 25cm, after 3 days cells separated by tripsin, counted with trepan blue and hemocytometer, divided into two density [high 200000] and [low 20000 cells]. Before transplantation of cells, six well plates coated with poly L lysin and inactivated astrocyte. Then isolated cells transplanted into 6 well plate for 4 days with medium DMEM/F12 supplemented with FBS10%. After 4 days different doses of ARTA and RA cis-9 added per well for 6 days, and then immunocytochemistery were done. After 6 days of treatment with above factors, doses of 100nM RA cis-9 and 500 nM ATRA have the more staining cells with monoclonal antibody. But in 100 nM RA cis-9, we saw maximum differentiated cells. All of differentiation were done on wells with inactivated astrocyte layer in high and low density. Inactivated astrocyte as a feeder layer and extrinsic factors such as All Transe Retinoic Acid [ATRA] and RA cis-9 can cause differentiation in hippocampal stem cells into photoreceptor like cell

[Effects of retinoic acid and astrocyte as a feeder layer in differentiation of hippocampal stem cell]

Background: Adult stem cells are pluripotent cells conventionally isolated from some part of body by different methods. From developmental stand point, murine neural stem cells represent an accessible and important system for studies of basic stem cell property such as self- renewal and multipotency

Materials and Methods: In this study hipocampal stem cells obtained from embryonic day 18 [E18] Pregnant female rats were killed, embryos heads separate and then hippocampus isolated by the method of Banker, then their cells dissociated by the methods of Fishbach, and plated in flask 25cm, after 3 days cells separated by tripsin, counted with trepan blue and hemocytometer, divided into two density [high 200000] and [low 20000 cells]. Before transplantation of cells, six well plates coated with poly L lysin and inactivated astrocyte, Then isolated cells transplanted into 6 well plate for 4 days with medium DMEM/F12 supplemented with FBS10%. After 4 days different doses of ARTA and RA cis-9 added per well for 6 days, and then immunocytochemistery were done

Results: After 6 days of treatment with above factors, doses of 100nM RA cis-9 and 500 nM ATRA have the more staining cells with monoclonal antibody . But in 100 nM RA cis-9, we saw maximum differentiated cells. All of differentiation were done on wells with inactivated astrocyte layer in high and low density

Conclusions: Inactivated astrocyte as a feeder layer and extrinsic factors such as All Transe Retinoic Acid [ATRA] and RA cis-9 can cause differentiation in hippocampal stem cells into photoreceptor like cell