The effects of transdermal estrogen delivery on bone mineral density in postmenopausal women: a meta-analysis

Due to its minimal systematic adverse effects, transdermal estrogen is widely used for the prevention of osteoporosis in postmenopausal women. The present meta-analysis aimed to clarify the effects of transdermal estrogen on bone mineral density [BMD] of postmenopausal women. Studies were identified by searching electronic databases including Cochrane Library, MEDLINE, Embase, and CINAHL databases, and also the Sciences Citation Index. Systematic review of articles was published between January 1989 to February 2016.Reference lists of the included articles were also evaluated and consultations were made with relevant experts. While 132 studies included the desired keywords, only nine clinical trials met the inclusion criteria and were finally reviewed. The pooled percent change in BMD was statistically significant in favor of transdermal estrogen. According to resulting pooled estimate, lumbar spine BMD one and two years after transdermal estrogen therapy was respectively 3.4% [95% CI: 1.7-5.1] and 3.7% [95% CI: 1.7-5.7] higher than the baseline values. The test for heterogeneity was not statistically significant based on the I2 heterogeneity index. One-two years of transdermal estrogen delivery can effectively increase BMD and protect the bone structure in postmenopausal women

Syphacia obvelata: A New Hope to Induction of Intestinal Immunological Tolerance in C57BL/6 Mice

The ability of nematodes to manipulate the immune system of their host towards a Th2 and T regulatory responses has been proposed to suppress the inflammatory response. Clinical trials have proposed a useful effect of helminth infections on improvement of inflammatory disorders. In this study, we investigated the immunomodulatory effect of Syphacia obvelata infection to induce intestinal tolerance in C57BL/6 mice. Mice were infected through the cagemates with self-infected BALB/c mice. Four weeks post-infection, expression levels of IFN-γ, TNF-α, IL-17, and IL-10 were assessed in the supernatant of mesenteric lymph node (MLN) culture. Foxp3⁺Treg were measured in MLN cells by flow cytometry. In the S. obvelata-infected group, the percentage of Tregs (5.2±0.4) was significantly higher than the control (3.6±0.5) (P<0.05). The levels of IL-10 (55.3±2.2 vs 35.2±3.2), IL-17 (52.9±3.8 vs 41±1.8), IFN-γ (44.8±4.8 vs 22.3±2.3) and TNF-α (71.1±5.8 vs 60.1±3.3) were significantly increased in infected mice compared to the control group (P<0.05). The above results showed the potential effects of S. obvelata to induce intestinal tolerance. Therefore, it seems that S. obvelata may increase the immunological suppressive function in the intestinal tract.

Effects of intraperitoneal administration of simvastatin in prevention of postoperative intra-abdominal adhesion formation in animal model of rat

Objective: To determine the preventive effects of local administration of simvastatin for postoperative intra-abdominal adhesion formation in animal model of rat

Methods: In this experimental study, 32 Wistar albino rats as the animal model of intra-abdominal adhesion formation were included. Adhesions were induced in all the animals via abrasion of the peritoneal and intestinal surface during laparotomy. Afterwards, the rats were randomly assigned to receive simvastatin [30 mg/kg body weight] as a single intraperitoneal dose at the time of laparotomy [n=16] or normal saline in same volume at the same time [n=16]. At the day 21, animals were euthanized and the adhesions were quantified clinically [via repeated laparotomy] and pathologically and compared between the two groups

Results: The baseline characteristics of the animals were comparable between two study groups. Clinically, in simvastatin group, 10 rats [62.5%] did not develop any adhesion and 6 [37.5%] had first-grade adhesion; whereas in the control group, 11 [68.8%] rats had first- and 5 [31.2%] had second-grade adhesions [p<0.001]. Pathologically, in simvastatin group, 6 rats [37.5%] had first-grade adhesion, while in control group, 11 rats [68.8%] had first- and 5 [31.2%] had second-grade adhesions [p<0.001]

Conclusion: Our findings suggest that intraperitoneal administration of simvastatin is an effective method for prevention of postoperative intra-abdominal adhesion formation in animal model of rat

Effect of olive leaf extract on cytokines secreted by macrophage

Background: herbal medicines in compared with chemical drugs have fewer side effects and can be a good medicinal alternative. The olive includes 20 different species of the family Oleaceae, with Olea europaea as the most recognized. Several studies have shown the immunomodulating effects of olive leaf extract. This study aimed to identify the immunoregulatory effect of olive leaf Sevillana variety on interleukins 12 and 10 which resulted from the murine macrophages in vitro

Materials and Methods: in order to isolate macrophages, peritoneal macrophages BALB/C were used. To determine the cytotoxic effect of different concentrations of the olive leaf extract on macrophages, MTT assay was performed. Concentrations of 200, 100, 50, 25, 12.5, 6.25, and 3.1[micro]g/ml in the time intervals of 12, 24, and 48 hours were evaluated. Three appropriate concentrations were selected to commence with the study of the determination of the amount of cytokines. Cell culture supernatant growth medium supernatant was collected at 12, 24, and 48 hours after adding the extract in order to examine the amount of cytokines. ELISA test was conducted using interleukins 12 and 10 measurement kits

Results: CC50 of the olive leaf extract at 12, 24, and 48 hours was 260.3, 170.5, and 150 [micro]g/ml, respectively. According to the results, an increase in the concentration and duration of the study resulted in observable significant differences in the production of interleukins 10 and 12. As a result, the production of IL-10 and 12 experienced decreases and increases, respectively

Conclusion: it seemed probable that the olive leaf extract had the capability to increase the production of IL-12 through activation of the classic macrophages and also deactivate the regulatory macrophages with an increase in IL-12 and a decrease in IL-10. Therefore, this can strengthen the immune system of the host in the early stages of infection. The other immunomodulatory effects of olive leaf must be considered by appropriate research

experimental model of colitis induced by dextran sulfate sodium from acute progresses to chronicity in C57BL/6: correlation between conditions of mice and the environment

Aim: To induce acute colitis progresses to chronicity in C57BL/6 mice by dextran sulfate sodium

Background: Murine models are essential tools to understand IBD pathogenesis. Among different types of chemically induced colitis models, the dextran sulfate sodium [DSS]-induced colitis model is the most common model of IBD, due to its simplicity

Patients and methods: Male C57BL/6 mice 6-8 weeks old, were collected and matched by age with controls. C57BL/6 mice treated with 2 cycles of 3.5% DSS for 4 days and 4 days of pure water between each cycle. After that, mice were sacrificed and the entire colon was removed. Small sections of the colon were fixed in formaldehyde, embedded in paraffin and sectioned with a microtome. Sections were stained with hematoxylin eosin to analyses the degree of inflammation

Results: After the first cycle oral administration of DSS, mice with severe and visible rectal bleeding and diarrhea entered into the acute phase. After day 4-5, bleeding and diarrhea were improved and mice entered into the chronic phase with peak levels of weight loss. Macroscopically, the inflammation was predominantly located in the distal colon. Microscopically, examination of the distal colon sections showed a decrease number of goblet cells, loss of crypts, signs of surface epithelial regeneration and moderate to severe infiltration of inflammatory cells in the mucosa

Conclusion: In order to achieve an experimental colitis model, our protocol is recommended for future therapies in IBD experimental modeling

Minocycline effects on IL-6 concentration in macrophage and microglial cells in a rat model of neuropathic pain

Background: Evidence indicates that neuropathic pain pathogenesis is not confined to changes in the activity of neuronal systems but involves interactions between neurons, inflammatory immune and immune-like glial cells. Substances released from immune cells during inflammation play an important role in development and maintenance of neuropathic pain. It has been found that minocycline suppresses the development of neuropathic pain. Here, we evaluated the analgesic effect of minocycline in a chronic constriction injury [CCI] model of neuropathic pain in rat and assessed IL-6 concentration from cultured macrophage and microglia cells

Methods: Male Wistar rat [n=6, 150-200 g] were divided into three different groups: 1] CCI+vehicle, 2] sham+vehicle, and 3] CCI+drug. Minocycline [10, 20, and 40 mg/kg] was injected one hour before surgery and continued daily to day 14 post ligation. Von Frey filaments and acetone, as pain behavioral tests, were used for mechanical allodynia and cold allodynia, respectively. Experiments were performed on day 0 [before surgery] and days 1, 3, 5, 7, 10, and 14 post -injury. At day 14, rats were killed and monocyte-derived macrophage from right ventricle and microglia from lumbar part of the spinal cord were isolated and cultured in RPMI and Leibovitz's media, respectively. IL-6 concentration was evaluated in cell culture supernatant after 24 h

Results: Minocycline [10, 20, and 40 mg/kg] attenuated pain behavior, and a decrease in IL-6 concentration was observed in immune cells compared to CCI vehicle-treated animals

Conclusion: Minocycline reduced pain behavior and decreased IL-6 concentration in macrophage and microglial cells

Nanomedicine Approaches for Treatment of Menopausal Symptoms

No abstract available.

Isolation and partial characterization of human amniotic epithelial cells: the effect of trypsin

Despite the extensive information available in the literature, cell surface marker signature of human Amniotic Epithelial Cells [hAECs] remains controversial. The aim of the present study was to characterize immunophenotypic features, proliferative capacity and immunogenicity of hAECs. We also tested whether expression of some cell surface markers is influenced by the type of trypsin used for tissue digestion. Single cell suspensions of amniotic membranes from four human placentas were isolated by enzymatic digestion and expression of CD9, CD10, CD29, CD34, CD38, CD44, CD45, CD73, CD105, CD133, HLA-I, HLA-DR, HLA-G, SSEA-4, STRO-1 and OCT-4 was then evaluated by flow cytometry. The differential impact of four trypsin types on the yield and expression of CD105 and HLA-I was also determined. The proliferative capacity of cultured hAECs was assessed and compared in the presence and absence of Epidermal Growth Factor [EGF]. To test their immunogenicity, hAECs were injected into Balb/c mice and the reactivity of hyperimmunized sera was examined by immunofluorescence staining. Nearly all purified cells expressed mesenchymal markers, CD9, CD10, CD29, and CD73 and the embryonic marker, SSEA-4. A large proportion of the cells also expressed STRO-1 and OCT-4. The purified cells also expressed HLA-G and HLA-I. A very small proportion of hAECs expressed CD34, CD38, CD44, CD133 and HLA-DR. The type of trypsin used for enzymatic digestion affected both the percentage and expression of HLA-I and CD105. hAECs revealed substantial proliferative capacity only when cultured in the medium supplemented with EGF. These cells were shown to be capable of inducing high amounts of anti-donor antibodies. Here we provided evidence that hAECs are immunogenic cells with high level of HLA-I expression. Furthermore, this work highlighted the impact of isolation procedure on the immunophenotype of hAEC

Histone modification of embryonic stem cells produced by somatic cell nuclear transfer and fertilized blastocysts

Nuclear transfer-embryonic stem cells [NT-ESCs] are genetically identical to the donor's cells; provide a renewable source of tissue for replacement, and therefore, decrease the risk of immune rejection. Trichostatin A [TSA] as a histone deacetylase inhibitor [HDACi] plays an important role in the reorganization of the genome and epigenetic changes. In this study, we examined whether TSA treatment after somatic cell nuclear transfer [SCNT] can improve the developmental rate of embryos and establishment rate of NT-ESCs line, as well as whether TSA treatment can improve histone modification in NT-ESCs lines. In this experimental study, mature oocytes were recovered from BDF1 [C57BL/6xDBA/2] mice and enucleated by micromanipulator. Cumulus cells were injected into enucleated oocytes as donor. Reconstructed embryos were activated in the presence or absence of TSA and cultured for 5 days. Blastocysts were transferred on inactive mouse embryonic fibroblasts [MEF], so ESCs lines were established. ESCs markers were evaluated by reverse transcription-polymerase chain reaction [RT-PCR]. Histone modifications were analyzed by enzyme linked immunosorbent assay [ELISA]. Result of this study showed that TSA treatment after SCNT can improve developmental rate of embryos [21.12 +/- 3.56 vs.8.08 +/- 7.92], as well as establishment rate of NT-ESCs line [25 vs.12.5]. We established 6 NT-ESCs in two experimental groups, and three embryonic stem cells [ESCs] lines as control group. TSA treatment has no effect in H3K4 acetylation and H3K9 tri-methylation in ESCs. TSA plays a key role in the developmental rate of embryos, establishment rate of ESC lines after SCNT, and regulation of histone modification in NT-ESCs, in a manner similar to that of ESCs established from normal blastocysts

Human Bhsp90 as adjuvant in HCV recombinant vaccine

There are more than 350 million individuals with hepatitis C in the world. One of the important problems in vaccine project is development of effective and suitable adjuvant in human vaccines. At present research we applied human BHsp90 protein as an adjuvant in recombinant HCV vaccine design. The thermal vector of pGP1-2 was used for human heat shock protein 90 expression. This protein injected to BalbC mice as an adjuvant together with recombinant protein of HCV core. The combination of these proteins was used and we evaluated the humoral and cellular immunity and the cytokine secretion of inguinal and popliteal lymph nodes lymphocytes were analyzed in vitro and ex vivo conditions. So the combination of Core protein together with hsp90 induced total IgG and IgG2a secretion. The spleen lymphocytes proliferation were increased equal to serum IgG2a level that was constant in second time bleeding with significant different to complexes with freund's adjuvant. At first IL-4 and IL-5 cytokines were increased, after one week it decreased. Production of IL-4 showed there was no hypersensitivity reaction after vaccine injection

Cytoplasmic and membranous CD24 marker expression has indirect correlation with cAMP/cGMP ratio

CD24 is a cell adhesion molecule that has been implicated in metastatic tumor progression cells. Our aim was to clarify the correlation between CD24 expression and cAMPxGMP ratio in murine colorectal cancer cell line [CT26] after using cholera toxin. The CT26 cells were cultured in flasks for assaying cAMP and cGMP by ELISA kits; also the cells were cultured in flasks for assaying cytoplasmic and membranous CD24 expression. The Real-Time PCR was done for cDNA that was synthesized from CT26 cells' mRNA. Also, expression CD24 marker of cells was determined by Anti-CD24 antibody and Goat Anti-Rabbit IgG-FITC [flow cytometry]. The cholera toxin increases the cAMPrcGMP ratio and influenced the cytoplasmic and membrane CD24 expression level. Cholera toxin increased cAMP. After increasing cAMP/cGMP ratio, cytoplasmic and membranous CD24 expression showed decline

[ study of cell viability and alkaline phosphatase activity of saos-2 cells contact with osteon and cerasoeb in vitro]

Osteon has been introduced as a bone substitute material. It is biphasic calcium phosphate [BCP] that use in dentistry. The aim of the present study was to evaluate the effect of osteon on the proliferation, cell viability and differentiation of saos-2 cells in vitro. Also it was compared with cerasorb bone graft. Two different bone grafts materials, osteon and cerasorb, were used to evaluate the effect on proliferation and differentiation rate of saos-2 cells. On day 15 the cell proliferation and cell viability was measured by MTT assay. For determination of differentiation, alkaline phosphatase and alizarin red test was used. Osteon and cerasorb groups showed significantly higher alkaline phosphatase activity and cell differentiation. Cell viability of both bone grafts was significantly lower than control group and cell proliferation was higher in osteon group. Osteon has more suitable biological property compare to cerasorb. The results of the present study showed that osteon and cersorb bone grafts allow proliferation and differentiation of saos-2 cells in vitro

[Effect of biomaterial grafts of Totudent and Bio-oss on mitochondrial activity and alkaline phosphatase levels of Saos-2 osteoblastoid cells]

Several reports on clinical utility of various Bone Substitute Materials, [BSM] for grafting have shown successful results. The most important factor in successful grafting of BSM is the physiological and histological behavior of this material which can be evaluated by observing, rate of survival and proliferation of osteogenic cells. This research was conducted to examine the effect of Totudent and Bio-oss on the morphology, proliferation and rate of survival of Saos-2 osteoblastoid cells in vitro. This experimental study was done on two types of materials used as bone grafts viz. Totudent and Bio-oss. Twelve samples from each of these 2 materials grafted on Saos-2 osteoblastoid cells and 4 negative control samples were studied. On day 15, the rate of cell survival was determined by the MTT test and on day 20, Alkaline phosphatase activity of these cells in the culture medium were evaluated by the Alkaline phosphatase kit. The rate of cell survival in the Bio-oss and Tutodent group was significantly diminished in comparison with the control group. This rate was significantly higher in the cells grafted with Tutodent as compared to Bio-oss; alkaline phosphatase activity in Totudent group was higher than the other 2 groups. Tutodent bone graft showed better result in the morphologic variations of Saos-2 cells in the culture medium as compared to Bio-oss, and has more appropriate biologic characteristics

[Comparing bone marrow mesenchymal cells and mesothelial cells of peritoneal fluid in expressing molecules of major histocompatibility complex [MHC]]

Mesothelium is composed of a single layer of mesothelial cells attached to a thin basement membrane supported by subserosal connective tissue; it plays an important role in homeostasis, wound healing, fluid transport and inflammation. The introduction of peritoneal dialysis [PD] as a modality of renal replacement therapy has provoked much interest in the biology of peritoneal mesothelial cell. This study was conducted to investigate the nature and plasticity of mesothelial cells isolated from peritoneal dialysed fluid [PDF], from patients on dialysis. Cell populations from peritoneal fluid of dialysed patients showing no evidence of peritonitis, and mesenchymal cells isolated from the bone marrow were cultured and screened for dominant surface markers by cell culture and immunofluorescence; MHC II, [DR, DQ], and MHC I were detected by flow cytometry method. Our results showed both bone marrow mesenchymal stem cells and mesenchymal cells obtained from peritoneal fluid do not express MHC II; also, peritoneal mesothelial cells have limited expression of MHC I. The use of peritoneal mesothelium as an option for grafting and regenerative therapy needs to be investigated in intensive research

Soluble Fas in pemphigus vulgaris

The Fas/Fas ligand [FasL] system has been recognized as an important pathway for apoptosis induction in cells and tissues. It has recently been shown that skin lesions of pemphigus vulgaris are associated with Fas mediated apoptosis. The aim of this study was to evaluated the level of serum soluble Fas of ten newly diagnosed patients with pemphigus vulgaris. Sera were collected from ten patients with pemphigus vulgaris. Commercial sandwich enzyme-linked immunosorbent assay [ELISA] for quantitative detection of soluble Fas was applied. Patients with mucosal skin involvement had higher median values in contrast to patients with cutaneous involvement. Elevation of soluble Fas in our study may give insights for the pathogenesis of pemphigus vulgaris. Suppression of this underlying mechanism may be an important target for novel therapies and relapse prevention

Lumbar puncture in neonates under and over 72 hours of age

To identify the criteria for performing a spinal tap in neonates with suspected sepsis and compare the findings in infants under and over the age of 72 hours. Analytical. Neonatal Ward of Taleghani Medical Centre, Shaheed Beheshti University of Medical Sciences, Evin,Tehran. Study period was of one year, from March 2003 to February 2004. All newborns subjected to a lumbar puncture during the first 24 hours of admission were enrolled in the study. Results of cerebrospinal fluid analysis were scrutinized. There were 380 neonates in the study. Majority i.e. 248 infants were aged < 72 hours. Meningitis was diagnosed in 22 cases, [5.8%]. There was no significant difference in the incidence of meningitis between neonates under and over 72 hours of age, [6.9% vs. 3.8%, p-value >0.5]. Comparison of risk factors, clinical manifestations, and routine laboratory investigations revealed no significant difference in babies with or without meningitis. The findings of this series do not support a selective approach for performing a lumbar puncture in a possibly septic newborn. A lumbar puncture is recommended in all neonates with suspected sepsis regardless of age, risk factors, or specific neurological manifestations

Release of sFasL by monocytes and lymphocytes triggered by betaglucan and zymosan

Polysaccharides have long been used as immune-modulators in various pathologic conditions including inflammation and solid malignancies. To evaluate the effects of Zymosan and Betaglucan on cytotoxic reactions in an effectortarget conjugate system. Blood was obtained from 20 healthy subjects; purified mononuclear leukocytes [monocytes and lymphocytes] were extracted and cultured as effector cells by a cytotoxic method. Both adherent and non-adherent cells interacted with the K562 myeloid cell line. The effector-target [E:T] ratio was 1:1, 1:10, and 1:20. To evaluate stimulatory effects of Betaglucan and Zymosan on cytotoxic reactions, samples were divided into case and control groups based on the presence or absence of Betaglucan and Zymosan. MTT assay and sFas ligand [sFasL] concentrations were used to assess the increased killing capacity of effector cells. Our results revealed that Zymosan and Betaglucan can induce cytotoxic responses in macrophages and lymphocytes [P<0.05]. The best result was achieved with E:T ratio of 1:1. Both macrophages and lymphocytes produced sFasL following stimulation by Zymosan and Betaglucan, however, the level of production was not statistically significant [P>0.05]. Zymosan and Betaglucan can be used as enhancers of the killing capacity of the immune cells; therefore, Betaglucan and Zymosan can be applied as systemic stimulators of the immune response in inflammation and chronic infection