Antidepressant effects of a Persian herbal formula on mice with chronic unpredictable mild stress.

Objective: Depression is a serious mental disorder. Despite numerous medications, there are still limitations in depression treatment. So, herbal medicine has been considered an alternative therapy. This survey evaluated the effects of a Persian herbal formula on mice with chronic unpredictable mild stress (CUMS). Materials and Methods: A combination of Aloysia triphylla citrodora, Citrus aurantium, Echium amoneum, Lavandula angustifolia, Melissa officinalis, Salix aegyptiaca, Valeriana officinalis, Viola odorata, and Cinnamomum zeylanicum was prepared. Except for the control group, animals were subjected to CUMS for 8 weeks in 5 groups (n=10): CUMS, vehicle (distilled water), herbal formula (0.23 ml/mouse), fluoxetine (20 mg/kg), and bupropion (15 mg/kg). All administrations were performed orally daily for the last 4 weeks. The depression and anxiety-like behaviors were assessed by sucrose preference (SPT), tail suspension (TST), forced swimming (FST), and elevated plus-maze (EPM) tests. Superoxidase-dismutase (SOD) activities in tissues, and serum levels of cortisol, alanine-aminotransferase (ALT), and creatinine were measured. Also, histopathological changes were evaluated. Results: This formula significantly increased SPT (p<0.001) and decreased immobility time in FST and TST (p<0.01), but it was not effective on EPM vs. CUMS mice. The herbal formula did not change the serum level of creatinine or ALT, but insignificantly reduced cortisol vs. CUMS and vehicle groups. SOD activity increased in the brain vs. vehicle group (p<0.05). There were no changes in histological examination. Conclusion: The herbal formula improved depression-like behaviors which are possibly related to its anti-oxidative effect on the brain. Also, it did not cause any negative changes in the biochemical and histopathological analysis.

Isolation of cellulose nanofibers from rapeseed straw via chlorine-free purification method and its application as reinforcing agent in carboxymethyl cellulose-based films.

In this study, cellulose nanofibers (CNFs) were successfully isolated from rapeseed straw (RS) whose valorization has been rarely investigated to date. A combined bleaching method without chlorine was applied for the purification of cellulose fibers, previously unexplored for RS. Chemical composition analysis and Fourier-transform infrared spectroscopy (FTIR) indicated that the purification method eliminated hemicellulose and reduced lignin content from 24.4 % to 1.8 %. The isolation of CNFs was performed using sulfuric acid hydrolysis under different acid concentrations (55 and 60 % v/v) and hydrolysis times (15, 30, and 45 min). The isolated CNFs were characterized by FTIR, X-ray diffraction (XRD), scanning electron microscopy (SEM), and thermogravimetric analysis (TGA). The formation of CNFs was confirmed by a significant increase in crystallinity index from 46.45 % of RS to >79.41 % of CNFs, depending on acid concentration and isolation duration. Carboxymethyl cellulose (CMC) films with different contents of CNFs were prepared by casting method. The mechanical properties and cytotoxicity of the prepared films were investigated. The CNFs obtained from RS via a chlorine-free purification method showed promising results for their usage as reinforcement in CMC matrix and film fabrication for various applications such as transdermal medicine and food packaging.

WITHDRAWN: The Sub-chronic Administration of Echium Oil Attenuates the Seizure Threshold and Improves Antioxidant Activity in Pentylenetetrazol- induced Seizure Model of Mice

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Therapeutic efficacy of triclabendazole in comparison to combination of triclabendazole and levamisole in sheep naturally infected with Fasciola sp.

Fascioliasis is an important parasitic disease which affects productivity of ruminants, and imposes significant economic losses. Benzimidazoles are effective in the treatment of fascioliasis; however, there are several reports on benzimidazoles resistant flukes. Combinational therapy is an approach to delay the emergence of resistant flukes. The present study aimed to compare the efficacy of triclabendazole (TBZ) and combination of triclabendazole and levamisole (TBZ + LVM) in the treatment of sheep naturally infected with Fasciola sp. For this purpose, 40 ewes infected with Fasciola sp. in three groups received TBZ, and TBZ + LVM, or remained untreated as CON. Fecal egg count (FEC), fecal egg count reduction (FECR), liver enzymes activity, albumin, globulin, and total protein levels were measured on day 0, 7, 14, and 28 post treatments. Obtained results showed that treatment with TBZ and TBZ + LVM resulted in significant reduction in FEC (P < 0.05), and FECR reached to values of higher than 90% on 28 day post treatment. The FEC for TBZ + LMV on day 7 and 14 were 12.25 ± 3.82 and 3.08 ± 1.03, respectively which was significantly lower in comparison to TBZ and CON (P < 0.05). Efficacy of TBZ + LMV was higher than TBZ on day 7 and 14 post treatment; however, no significant difference was observed on 28 day. The liver enzyme activities on days 7 and 14 were lower in the TBZ + LVM sheep in comparison to the TBZ and CON. Treatment with TBZ or TBZ + LVM resulted in an increase in albumin and a decrease in globulin. Over all, the present study clarified the importance of combinational therapy, and demonstrated that combination of TBZ + LVM resulted in higher efficacy and earlier improvement of liver conditions in sheep naturally infected with Fasciola sp.

Differences between some biochemical components in seminal plasma of first and second ejaculations in dual-purpose Simmental (Fleckvieh) bulls and their relationships with semen quality parameters.

This study aimed to evaluate differences in seminal plasma zinc (Zn), copper (Cu), iron (Fe), catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx) levels in the first and second ejaculations and their relationships with semen quality parameters in Fleckvieh bulls. Repetitive ejaculates were separately collected, analyzed, and frozen from the sires. Progressive motility of frozen-thawed semen (PMFT) was considered the main factor for more data classification into three following groups: <40.00%, 40.00 - 50.00%, and >50.00%. Seminal plasma trace elements and enzymes were determined using atomic absorption spectrometry and ELISA, respectively. The results revealed significant differences between the first and second ejaculations. Semen concentration, SOD, GPx, and Fe were different in ejaculations. Although PMFT groups in different ejaculations did not show significant differences, there was significant alteration between different PMFT groups and first and second ejaculations. All frozen-thawed semen CASA parameters (except lateral head displacement) were associated with fresh motility parameters and before and after thawing sperm viability. Also, a correlation between seminal Zn concentration with fresh semen gross and progressive motility, average path velocity, and beat cross frequency, Cu with SOD and Fe and semen concentration was observed. CAT was associated with fresh and frozen-thawed sperm motility parameters except for lateral head displacement and angular displacement. Although our findings showed differences between the first and second ejaculations in some parameters, PMFT, which is the most important indicator for estimating bull fertility, was not different between them.

Effect of docosahexaenoic acid plus insulin on atherosclerotic human endothelial cells.

BACKGROUND: Atherosclerosis is touted as one of the most critical consequences of diabetes mellitus indicated by local inflammation of endothelial cells. The Effect of Omega 3 fatty acids, mainly docosahexaenoic acid (DHA), has been investigated in cells after exposure to high doses of lipids. The current experiment aimed to address the modulatory effects of docosahexaenoic acid and insulin in palmitic-treated human endothelial cells. METHODS: Human umbilical vein endothelial cells were treated with 1 mM palmitic acid, 50 µM insulin, 50 µM docosahexaenoic acid, and their combination for 48 h. Cell survival rate and apoptosis were measured using MTT and flow cytometry assays. The Griess assay detected NO levels. Protein levels of TNF-α, IL-6, and NF-κB were studied using ELISA and immunofluorescence imaging. The expression of genes participating in atherosclerosis was monitored using PCR array analysis. RESULTS: Oil Red O staining showed the inhibitory effect of DHA and insulin to reduce the intracellular accumulation of palmitic acid. Both DHA and Insulin blunted palmitic acid detrimental effects on HUVECs indicated by an increased survival rate (p < 0.05). The percent of apoptotic cells was decreased in palmitic-treated cells received insulin and DHA compared to palmitic-treated group (p < 0.05). Based on our data, DHA and Insulin diminished the production of all inflammatory cytokines, TNF-α, IL-6, and NF-κB, in palmitic-treated cells (p < 0.05). Similar to these data, NO production was also decreased in all groups treated with insulin and DHA compared to the palmitic-treated cells (p < 0.05). PCR array analysis revealed the modulatory effect of DHA and insulin on the expression of atherosclerosis-related genes pre-treated with palmitic acid compared to the control group (p < 0.05). CONCLUSION: DHA and Insulin could alter the dynamic growth and dysfunctional activity of human endothelial cells after treatment with palmitic acid. Taken together, Omega 3 fatty acids, along with insulin, could dictate specific cell behavior in endothelial cells in vitro.

The Toxicity Effect of Echium amoenum on the Liver and Kidney of Mice.

AIMS: The aim of this study was to investigate the toxic effect of Echium amoenum plants on the liver and kidney of the animal model. BACKGROUND: Echium amoenum is one of the medicinal plants containing pyrrolizidine alkaloids with several properties which has widely consumed among different communities. OBJECTIVE: The toxic effects of Echium amoenum on the liver and kidney were investigated in this study. METHODS: Sixty mice were kept for 28 days under the appropriate laboratory conditions. Echium amoenum extract (25, 12.5, 50 mg / kg, ip.) was administered for 28 days. At the end of the experiment, blood samples were drawn and liver and kidneys were removed for evaluating hepatotoxicity and nephrotoxicity of extract. Additionally, experiments were conducted to assay the enzymatic and oxidative activities. RESULTS: There was no significant difference in the levels of copper ion in the liver and kidneys among all groups. There was a significant difference in the levels of lipid peroxidation in the liver of treated groups versus the control group. The significant difference was not observed in the levels of glutathione of the liver of all groups. However, the levels of glutathione of the kidney significantly decreased in the treated groups versus the control group. There was no significant difference in the liver enzymes, including ALP, SGOT, and SGPT, between all groups. This indicates that damage increases with enhancing the time and concentrations of the extract. Biochemical analysis showed the creatinine and urea levels did not change in the treated groups versus the control group. CONCLUSION: According to the present findings, it is suggested that Echium amoenum causes hepatotoxicity and nephrotoxicity effects in dose and time-dependent manner.

Antileishmanial activity of Urtica dioica extract against zoonotic cutaneous leishmaniasis.

BACKGROUND: Neglected parasitic diseases (NTDs) like cutaneous leishmaniasis (CL) have caused high mortality and morbidity rate in developing countries. This disease is considered as one of the six major tropical diseases, and has a great importance in HIV infected individuals as an opportunistic infection in those areas that both infections are endemic. This study evaluated the therapeutic effects of the Urtica dioica L (U. dioica) aqueous extract as an anti-leishmanial herbal drug in-vitro and in-vivo, and in addition to that, evaluated two vital immune system cytokines including gamma interferon (IFN-γ) and interleukin-4 (IL-4) plus nitric oxide (NO) and arginase activity against Leishmania major (L. major) infected mice. METHODOLOGY/PRINCIPAL FINDINGS: In-vitro anti-leishmanial activity of U. dioica aqueous extract was determined using MTT method and also Parasite Rescue Transformation Assay. Also, the footpad lesion size and parasite load in BALB/c mice infected with L. major were quantified for in-vivo assessment. Furthermore, for evaluating the immune responses, the levels of IFN-γ, IL-4, NO and arginase were measured in the BALB/c mice. These results indicated that U. dioica extract significantly reduced the L. major promastigotes viability. According to the in-vitro cytotoxicity assay of the extract on Leishmania parasites (CC50) and infected macrophages (EC50), the extract had no toxicity to the macrophages, however it efficiently killed the L. major amastigotes. In addition, the lesion size, parasite load, IL-4, and ARG were decreased in the treated infected mice, however IFN-γ and NO were significantly increased. CONCLUSIONS/SIGNIFICANCE: This study established satisfactory results in Leishmania parasite clearing both in-vivo and in-vitro. Therefore, U. dioica extract can be considered as an effective and harmless herbal compound for killing the parasite without toxicity to the host macrophages. Furthermore, it also can treat the CL by switching the mouse immune response towards a cell-mediated response (Th1); hence, it may be identified as a perfect therapeutic herbal drug for CL treatment.

Protective effects of 2-methoxycinnamaldehyde an active ingredients of Cinnamomum cassia on warm hepatic ischemia reperfusion injury in rat model.

OBJECTIVES: Hepatic ischemia/reperfusion injury (IRI) is one of the major causes of hepatic failure during liver transplantation, trauma, and infections. The present study investigated the protective effect of intra-portal administration of 2-methoxycinnamaldehyde (2-MCA) on hepatic IRI in rats. MATERIALS AND METHODS: Twenty-four rats were equally divided into four groups; 1) sham group, (no IRI or transfusion), 2) Hepatic IRI (60 min ischemia + 120 min reperfusion, 3) Hepatic IRI+ NS (IRI + normal saline), 4) Hepatic IRI+2-MCA, (IRI + 2-MCA). In groups 3 and 4, 1 ml/kg normal saline and 2-MCA were administered slowly into the vein of the left lateral and median lobes of the liver 10 min before induction of hepatic reperfusion (upper the site of clumping), respectively. The harvest time points were at 2 hours post-reperfusion in all groups. RESULTS: Histologically, cell death, degenerative changes, sinusoidal dilatation, congestion, hemorrhage, and infiltration of inflammatory cells were observed in IRI group, while these pathological changes were attenuated in the 2-MCA administrated group. The level of alanine transaminase, aspartate transaminase, tumor necrosis factor- α and interleukin-6 in serum and hepatic malondialdehyde were significantly increased by IRI, and 2-MCA administration reduced all these markers. In addition, caspase-3 and nuclear factor κB (NF-κB) expression were investigated immunohistochemically. Administration of 2-MCA considerably decreased caspase-3 positive cells and NF-κB activity in comparison with IRI group. CONCLUSION: As a conclusion, in situ administration of 2-MCA protects against hepatic IRI via anti-inflammatory, and anti-apoptotic properties.

Differences between first and second ejaculations regarding seminal plasma calcium, magnesium and total antioxidant capacity in dual-purpose Fleckvieh bulls and their relationships with semen quality.

Artificial insemination is a well-established and widely used method for genetic improvement in cattle breeding industry. Recently, researchers have shown an increased interest in the cryoprotective effects of minerals and antioxidants on semen. Previous studies on calcium (Ca) and magnesium (Mg), two main macro-minerals, have mainly investigated their roles in mammalian spermatogenesis and fertility. In addition, the experimental data examining the semen content regarding these minerals and antioxidants from different animal species are rather controversial and there is no general agreement about their associations with semen quality. Therefore, this study was conducted to assess the seminal plasma concentrations of Ca, Mg and total antioxidant capacity (TAC) in first and second ejaculations of dual-purpose Fleckvieh bulls and to link them to the sperm characteristics of fresh and frozen-thawed semen. Sperm progressive motility after thawing was used to classify the data into three groups: < 40.00%, 40.00 to 50.00% and > 50.00%. The measurements of two minerals and TAC were carried out using spectrophotometry and enzyme-linked immunosorbent assays, respectively. The results showed that there were significant differences in several parameters of semen quality between first and second ejaculations. No significant differences were also found on Ca and Mg concentrations and Ca/Mg ratio. The TAC level was significantly higher in the first ejaculation than the second one. The findings of this study suggest that TAC is a potential marker for bull semen quality assessment in the frozen semen production industry.

The Effect of Borage (Echium amoenum) on the Mouse Heart and Hematology Parameters.

BACKGROUND: There has been considerable interest in the potential health benefits of borage. Little information is available regarding the safety of this plant. The purpose of this study was to evaluate the impact of borage on the mouse heart. METHODS: Different amounts of borage extract were injected in mice. The mice were randomly divided into 4 groups including group1 (Control group without injection), group2, 3 and 4 that received 12.5 mg/kg, 25 mg/kg and 50 mg/kg respectively for 28 days. Oxidative stress parameters (lipid peroxidation, total glutathione groups assay and cupric assay) and biochemical (Creatine kinase activity and total cholesterol) and hematology parameters were evaluated. Furthermore, histopathology study was carried out on heart tissues. RESULTS: We found that there was no significant difference in oxidative stress parameters and biochemical parameters between the control group and the groups that received different amounts of borage extract. There were also no changes in histopathology study. In blood parameters, the level of erythrocytes, hematocrit and hemoglobin decreased to 50mg/kg, whereas the level of MCH and MCV decreased in high doses. CONCLUSION: This article suggested that borage did not cause significant damage to the heart tissue in mice model. In hematology factors, significant changes were observed in erythrocytes and related parameters. Therefore, hematotoxicity of consumption this plant should be considered at high doses.

Carvacrol promotes angiogenic paracrine potential and endothelial differentiation of human mesenchymal stem cells at low concentrations.

OBJECTIVES: Phenolic monoterpene compound, named Carvacrol, has been found to exert different biological outcomes. It has been accepted that the angiogenic activity of human mesenchymal stem cells was crucial in the pursuit of appropriate regeneration. In the current experiment, we investigated the contribution of Carvacrol on the angiogenic behavior of primary human mesenchymal stem cells. METHODS: Mesenchymal stem cells were exposed to Carvacrol in a dose ranging from 25 to 200µM for 48h. We measured cell survival rate by MTT assay and migration rate by a scratch test. The oxidative status was monitored by measuring SOD, GPx activity. The endothelial differentiation was studied by evaluating the level of VE-cadherin and vWF by real-time PCR and ELISA analyses. The content of VEGF and tubulogenesis behavior was monitored in vitro. We also conducted Matrigel plug in vivo CAM assay to assess the angiogenic potential of conditioned media from human mesenchymal stem cells after exposure to Carvacrol. RESULTS: Carvacrol was able to increase mesenchymal stem cell survival and migration rate (p<0.05). An increased activity of SOD was obtained while GPx activity unchanged or reduced. We confirmed the endothelial differentiation of stem cells by detecting vWF and VE-cadherin expression (p<0.05). The VEGF expression was increased and mesenchymal stem cells conditioned media improved angiogenesis tube formation in vitro (p<0.05). Moreover, histological analysis revealed an enhanced microvascular density at the site of Matrigel plug in CAM assay. CONCLUSIONS: Our data shed lights on the possibility of a Carvacrol to induce angiogenesis in human mesenchymal stem cells by modulating cell differentiation and paracrine angiogenic response.

In vivo evaluation of toxicity and antiviral activity of polyrhodanine nanoparticles by using the chicken embryo model.

Evaluation of the potential cytotoxicity of polyrhodanine nanoparticles is an important factor for its biological applications. In current study, for the first time histopathological and biochemical analysis of polyrhodanine besides of its antiviral activity against Newcastle disease virus (NDV) were examined on chicken embryo model. Polyrhodanine was synthesized by the chemical oxidative polymerization method. The obtained nanoparticles were characterized by scanning electron microscopy (SEM), and Fourier transform infrared (FTIR). Different doses of polyrhodanine nanoparticles were injected into the albumen in 4-day-old embryonic eggs for groups: (0.1ppm, 1ppm, 10ppm and 100ppm), while the Control group received only normal saline. The gross examination of chicks revealed no abnormality. No pathological changes were detected in microscopical examination of the liver, kidney, spleen, heart, bursa of Fabricius and central nervous system tissues. Blood serum biochemical indices showed no significant differences between control and treatment groups. Interestingly, polyrhodanine nanoparticles showed strong antiviral activity against NDV in ovo. These preliminary findings suggest that polyrhodanine nanoparticles without any toxicity effect could be utilized in controlling Newcastle disease in chickens.

Safety assessment of rice bran oil in a chicken embryo model.

OBJECTIVE: Rice Bran Oil (RBO) is extracted from the outer layer of rice. Little information is available regarding its safety. The present study was conducted to assess its safety in chicken embryo model. MATERIALS AND METHODS: RBO was injected on day 4 of incubation of chickens. The tissues and serum samples were collected. Oxidative stress parameters in the liver, kidney and brain and biochemical parameters of serum were measured. The deformities were also investigated. RESULTS: The changes in the liver enzymes activity were not statistically significant. There was significant decrease and increase in lipid peroxidation and glutathione level, respectively. It is suggested that RBO is a natural antioxidant source. Low-density lipoprotein cholesterol (LDL) also decreased. No abnormal findings were observed in the chickens. CONCLUSION: No toxic effect was observed following RBO administration in chicken embryos. This study showed that RBO is not a safety concern.

The Effect of Stevia on the Chicken Embryo Heart.

BACKGROUND: Stevioside is one of the most important food additives that has become well known for its sweetness. The aim of this study was to evaluate the impact of Stevioside on the heart. METHODS: 4-day-old embryonated chickens eggs were inoculated with Stevioside and kept until hatching. Shortly after, the heart tissue samples were taken to examine organ Oxidative stresses by measuring Malondialdehyde (MDA) and glutathione (GSH) levels, ferric reducing /antioxidant power (FRAP) and cupric ion reducing assay (CUPRIC). RESULTS: There was no significant difference in glutathione level, lipid peroxidation, FRAP, and cupric assay. CONCLUSION: It was suggested that stevioside did not cause marked damages to heart tissues in chicken embryo model.

Carvacrol as a potent natural acaricide against Dermanyssus gallinae.

Resistance to conventional synthetic pesticides has been widely reported in Dermanyssus gallinae in poultry production systems. Introducing novel acaricides to poultry industry today is more urgent than ever. Research in this field recently focused on plants and plant-derived compounds as acaricides. In the present study, acaricidal activity of three plant bioactive components, carvacrol, thymol, and farnesol, was assessed against D. gallinae and compared with synthetic pesticide permethrin. Mode of acaricidal action was determined by contact toxicity and fumigant toxicity bioassays. Except farnesol which did not cause any mortality, carvacrol and thymol were found to be toxic to D. gallinae with LD50 values of 1 and 3.15 µg/cm(3), respectively. Permethrin gave the LD50 value of 31.95 µg/cm(3) which was less efficient than carvacrol and thymol. In fumigant toxicity bioassay, mortality rate in carvacrol- and thymol-treated groups in closed method was significantly higher than the open one. On the other hand, permethrin exhibited poor fumigant toxicity as there was no statistically significant difference between mortality rate in open and closed methods. These findings revealed that mechanism of acaricidal activity of carvacrol and thymol but not permethrin was mainly due to fumigant action. Results of the present study suggested that carvacrol and thymol, especially carvacrol, can be developed as a novel potent bioacaricide against D. gallinae.

Endothelial juxtaposition of distinct adult stem cells activates angiogenesis signaling molecules in endothelial cells.

Efficacy of therapeutic angiogenesis needs a comprehensive understanding of endothelial cell (EC) function and biological factors and cells that interplay with ECs. Stem cells are considered the key components of pro- and anti-angiogenic milieu in a wide variety of physiopathological states, and interactions of EC-stem cells have been the subject of controversy in recent years. In this study, the potential effects of three tissue-specific adult stem cells, namely rat marrow-derived mesenchymal stem cells (rBMSCs), rat adipose-derived stem cells (rADSCs) and rat muscle-derived satellite cells (rSCs), on the endothelial activation of key angiogenic signaling molecules, including VEGF, Ang-2, VEGFR-2, Tie-2, and Tie2-pho, were investigated. Human umbilical vein endothelial cells (HUVECs) and rat lung microvascular endothelial cells (RLMECs) were cocultured with the stem cells or incubated with the stem cell-derived conditioned media on Matrigel. Following HUVEC-stem cell coculture, CD31-positive ECs were flow sorted and subjected to western blotting to analyze potential changes in the expression of the pro-angiogenic signaling molecules. Elongation and co-alignment of the stem cells were seen along the EC tubes in the EC-stem cell cocultures on Matrigel, with cell-to-cell dye communication in the EC-rBMSC cocultures. Moreover, rBMSCs and rADSCs significantly improved endothelial tubulogenesis in both juxtacrine and paracrine manners. These two latter stem cells dynamically up-regulated VEGF, Ang-2, VREGR-2, and Tie-2 but down-regulated Tie2-pho and the Tie2-pho/Tie-2 ratio in HUVECs. Induction of pro-angiogenic signaling in ECs by marrow- and adipose-derived MSCs further indicates the significance of stem cell milieu in angiogenesis dynamics.

Dynamic induction of pro-angiogenic milieu after transplantation of marrow-derived mesenchymal stem cells in experimental myocardial infarction.

BACKGROUND: Cell-based pro-angiogenic therapy by bone marrow mesenchymal stem cells (MSCs) has been touted as a means to reducing the adverse effects of cardiac remodeling after myocardial infarction (MI). Milieu-dependent regulation of pro-angiogenic potential of MSCs after infarction remains to be elucidated. In this study, the effects of marrow-derived MSCs on the kinetics of angiogenesis signaling factors were investigated in a rabbit model of MI. METHODS: MI was induced in rabbits, and the animals were randomized into two groups (cell transplantation and control, each group with 21 animals). 1 × 10(6) autologous marrow-derived MSCs were injected into the myocardium of the border zone after transfection with a green fluorescent protein (GFP) lentiviral reporter vector. Control animals received PBS vehicle only. Effect of the transplanted cells on the hearts was evaluated over time by pathological, immunofluorescence, western blotting, immuno-electron microscopy, and echocardiographic analyses. RESULTS: Transplanted GFP-positive MSCs were enriched with time in the peri-infarct border zone with differentiation potential into three major cell types of the heart, including cardiomyocytes, endothelial cells, and smooth muscle cells, and there was significant augmentation of microvascular density. The transplanted cells could change the milieu of the injured myocardium to increase the expression levels of VEGF as well as the ratio of Ang-2 to Ang-1, and to reduce the ratio of phosphorylated Tie2 to Tie2. CONCLUSION: An angiogenesis-promoting milieu was induced after the transplantation of marrow MSCs in the injured myocardium. Compared with the resident cells, the transplanted cells had a greater rate of cellular kinetics in the infarcted myocardium.

Flow cytometric immunophenotyping of feline bone marrow cells and haematopoietic progenitor cells using anti-human antibodies.

There is a paucity of species-specific antibodies available for feline haematopoietic conditions. The purpose of this study was to broaden the panel of antibodies available for use in the immunophenotypic characterisation of feline haematopoietic cells by testing clones of anti-human monoclonal antibodies (mAbs) on normal, neoplastic and cultured feline haematopoietic progenitors to determine cross-reactivity to feline counterparts. In this study, 24 clones of anti-human mAbs were tested on normal or neoplastic feline bone marrow and peripheral blood cells. Six of these mAbs, including anti-cluster of differentiation (CD)61, anti-CD18, anti-CD14, anti-CD235a, anti-CD41 and anti-CD29, cross-reacted with normal feline bone marrow cells, whereas anti-CD33 and anti-CD117 cross-reacted with the blast cells in the bone marrow of two cats with myelodysplastic syndrome, and anti-CD71, anti-235a, anti-41 and anti-42 cross-reacted with immature erythroid cells in a cat with erythroleukaemia. In a feline immunodeficiency virus-positive cat, bone marrow cells were labelled with anti-CD33, anti-14 and anti-45. Anti-CD18, anti-CD14, anti-CD41 and anti-CD61 also reacted with the peripheral blood cells of the healthy cats. The feline haematopoietic progenitors formed colonies in the methylcellulose-based semisolid medium with significant enrichment of colony-forming unit-granulocyte, monocyte and burst-forming unit-erythroid. A panel of six anti-feline mAbs (anti-CD21-like, anti-T lymphocytes, anti-CD172a, anti-granulocyte, anti-CD45-like and anti-CD18) and eight anti-human antibodies (anti-CD71, anti-CD33, anti-CD235a, anti-CD41, anti-CD61, anti-CD117, anti-CD38 and anti-CD34) were used for the immunophenotypic characterisation of the feline bone marrow progenitors. CD45, CD33, CD235a and CD18 were expressed by the feline haematopoietic progenitor cells, with the highest expression level for CD45.

Haemoglobin typing and its variations in Iranian domestic dogs.

The study of canine haemoglobin (Hb) components can help to forecast Hb changes during many pathological and physiological processes such as responsive anaemia. The aims of our study were to show canine Hb electrophoretic pattern on cellulose acetate and identify Hb types similar to the human Hb pattern. Blood samples from 78 different canine breeds were randomly collected in tubes containing anticoagulant EDTA. Animals were brought to the Small Animal Teaching Centre for a check-up and vaccination. All blood samples underwent electrophoresis on cellulose acetate paper strips to determine Hb types. Haematocrit and Hb were measured simultaneously. The Hb electrophoresis results showed that Hb A(1) was assigned to most of Hb components on cellulose acetate paper. Also, in some blood samples, Hb A(2) was detected at the cathode end of cellulose acetate paper similar to human Hb A(2), by densitometer. Small amounts of Hb F were detected in some dogs which was not significant. According to our study, Hb A(1) composes most of the total Hb percent in canine blood. Two types of Hb, A(2) and F, were detected in a few animals, but the majority did not possess Hb F. It seems that Hb F is not significant in these animals. We concluded that one or two Hb types could be seen in dogs. There is no difference in electrophoretic pattern between male and female dogs.