فهرست مطالب hassan malekinejad

Polycystic ovary syndrome (PCOS) is one of the main causes of infertility in women. This study was conducted to uncover the effects of lupeol as an anti-androgenic triterpene on experimentally-induced PCOS in mice. 

Eighty immature female mice were divided into 4 groups: Control (C), PCOS (P), Lupeol (L), and Flutamide (F). PCOS was induced in test groups by injection of Dehydroepiandrosterone (60 mg/kg/day, IP) for twenty days. Following the PCOS induction, the two groups of L and F were treated with lupeol (40 mg/kg/day) and/or flutamide (10 mg/kg/day) respectively and the two groups of C and P received sesame oil (0.1 ml/mouse/day) for 15 days. After the treatment period, ten animals in each group were selected for collecting blood and ovary samples. In vitro  fertilization assessment was carried out on 10 remaining mice in each group. The hormonal assays and oxidative stress biomarker determination were performed on serum and tissue samples. Moreover, histopathological analyses were conducted on the ovaries.

PCOS-elevated concentration of LH and Testosterone was significantly (P<0.05) lowered in lupeol and flutamide-received animals. Lupeol and flutamide both reduced PCOS-induced fibrosis and the number of atretic follicles. Both compounds declined the PCOS-increased lipid peroxidation and protein oxidation in serum and the ovaries. Lupeol increased the PCOS-reduced fertility rate and decreased the number of arrested embryos by 12%. 

These findings indicate that lupeol could be a novel compound in the treatment of PCOS as it reduced PCOS-induced structural and also functional disorders.

Bacterial nanocellulose is known as a potential carrier for a widespread spectrum of biological compounds, including antibacterial and antifungal compounds. The present study was conducted to determine the impact of bacterial nanocellulose containing Natamycin and Amphotericin B on Aspergillus flavus and Penicillium citrinum in an in vitro environment.

In this descriptive-analytical research, Aspergillus flavus-PTCC: 5006 and Penicillium citrinum-PTCC: 5304 fungi were prepared from the Fungal Collection of the Department of Microbiology, Faculty of Veterinary Medicine, Urmia University. The minimum inhibitory concentration (MIC) and the minimum fungicidal concentration (MFC) of Natamycin and Amphotericin B against Aspergillus flavus and Penicillium citrinum were evaluated by the microdilution method. Bacterial nanocellulose was prepared using Komagata xylinum bacterium, and Natamycin and Amphotericin B were added in three concentrations of 0.01%, 0.05%, and 0.1% to wet and lyophilized nanocellulose films by the immersion method. Then, the antifungal effects of the film containing the above compounds against the investigated fungi were investigated by the agar diffusion method. Parchment paper was used as a control for comparison. Spectral properties of nanocellulose film containing antifungal compounds were evaluated by the Fourier transform infrared (FTIR) method.

MIC and MFC of Natamycin for Aspergillus flavus were determined as 3.9 μg/mL and 7.81 μg/mL, and for Penicillium citrinum as 7.81 μg/mL and 15.62 μg/mL, respectively. MIC and MFC of Amphotericin B for Aspergillus flavus were determined as 7.81 μg/mL and 15.62 μg/mL, and for Penicillium citrinum as 15.62 μg/mL and 31.25 μg/mL, respectively. The increased concentration had a statistically significant impact on the antifungal properties of all films (P<0.05). The best antifungal effects of the film were related to the film containing Natamycin.

Bacterial nanocellulose containing Natamycin showed stronger antifungal effects in an in vitro environment compared to Amphotericin B against Aspergillus flavus and Penicillium citrinum.

The current study aimed to study the therapeutic effects of lupeol as a nutritional triterpene on non-alcoholic fatty liver disease (NAFLD) and polycystic ovarian syndrome (PCOS) disorders in separate and concurrent models.

Experimental approach: 

This study was performed in three sets and each set contained 4 groups of female mice (n = 6), including control, NAFLD or PCOS and/or NAFLD/PCOS, lupeol, and metformin (MET). The treatment groups following the induction of disorders were treated with lupeol (40 mg/kg, orally) or MET (500 mg/kg, orally) for 28 days. The insulin resistance index and hormonal assessments were conducted on the collected serum samples. Moreover, oxidative stress biomarkers were measured in the liver and ovaries. Histopathological studies and ultimately any changes in the expression of androgen receptors, toll-like receptor (TLR)-2 and TLR-4 were analyzed.

Results revealed that lupeol reduced significantly the insulin resistance index in NAFLD and NAFLD/PCOS-positive animals. Lupeol attenuated remarkably the PCOS and PCOS/NAFLD-elevated concentration of testosterone. lupeol recovered the metabolic disorders-induced oxidative stress and restored the disorders-depleted glutathione. The NAFLD/PCOS-induced hepatic damages such as microvesicular or macrovesicular steatosis and atretic follicles number in the ovary were attenuated in the lupeol-treated mice. Serum level of TNF-α was reduced and the expression of androgen receptors, TLR-4 and TLR-2 were downregulated in the lupeol-treated NAFLD/PCOS-positive animals.

Conclusions and implication: 

The results suggest that lupeol could be a novel nutraceutical for the treatment of metabolic disorders. Lupeol’s anti-metabolic disorders effects attribute to its anti-dyslipidemia, antioxidant, and anti-inflammatory properties.

The need of human society is drawn towards healthier and more useful foods and the food industry is expected to pay attention to this need. This study was conducted with the aim of adding encapsulated Lactobacillus Casei to rye drink and investigating the diversity of the bacterial population and intestinal histopathology in the rat animal model.

Lactobacillus Casei was encapsulated with a concentration of 2% sodium alginate polymer by emulsion method. After adding a standard dose of encapsulated bacteria to rye drink, in vivo experiments were performed on 4 groups of adult male rats as: control, malt drink, rye drink, and rye drink with probiotics. Animals received the recommended beverages for 28 days. Bacterial population diversity was done by molecular method (PCR-DGGE) and histopathological examination using staining (H&E).

There was bacterial diversity in the intestinal contents of the studied rats, and the highest population composition related to Lactobacillus Casei was detected in the intestinal contents of rats that received rye drink with probiotics. Also, histopathological studies did not have negative results in any of the groups.

The results of this research show that the use of microencapsulation technique can increase the shelf life of Lactobacillus Casei in the intestine.

Colonic anastomosis is associated with serious complications leading to significant morbidity and mortality. Fibroblasts have recently been introduced as a practical alternative to stem cells because of their differentiation capacity, anti-inflammatory, and regenerative properties. The aim of this study was to evaluate the effects of intramural injection of fibroblasts on the healing of colonic anastomosis in rats.

Inbred mature male Wistar rats were used in this experimental study (n=36). Fibroblasts were isolated from the axillary skin of a donor rat. In the sham group, manipulation on descending colon was done during laparotomy. A 5 mm segment of the colon was resected, and end-to-end anastomosis was performed. In the control group, 0.5 ml of phosphate buffer saline (PBS) was injected into the colonic wall and in the treatment group, 1×106 fibroblasts were transplanted. Following euthanasia on day 7, intra-abdominal adhesion, leakage and peritonitis were evaluated by necropsy. Mechanical properties were assessed using bursting pressure and tensile tests. Inflammation, angiogenesis, and collagen deposition were examined histopathologically.

The mean scores for adhesion and leakage were decreased in the treatment group versus control samples. Lower infiltration of inflammatory cells was observed in the treatment group (P=0.03). Angiogenesis and collagen deposition scores were significantly increased in the fibroblast transplanted group (P=0.03). Tensile mechanical properties of the colon were significantly increased in the treatment group compared to the control sample (P=0.01). There was no significant difference between the control and treatment groups in terms of bursting pressure (P=0.10). Positive weight changes were found in sham and treatment groups, but the control rats lost weight after 7 days.

The results suggested that allotransplantation of dermal fibroblasts could improve the necroscopic, histopathological, and biomechanical indices of colonic anastomosis repair in rats.

Drug resistance is a great concern worldwide and in Iran. This study was carried out to assess drug resistance of gastrointestinal nematodes to Albendazole (Alb) and Fenbendazole (Feb) in sheep.

A total number of 90 fresh fecal samples were directly collected from the rectum of infected sheep with an average egg per gram of feces (EPG)≤150. A dilution of 1000 eggs per 2mL was added to each well of control, Alb (0.1µg/ml) and Feb (0.1µg/ml) groups and incubated. To determine drug resistance, a lethal dose of 50% (LD50) was calculated based on percentage of the hatched eggs and first larvae stage (L1) counting.

EPG was lower in Feb treated groups (43.76±1.73) than Alb treated groups (65.51±1.4) and control group (93.96±0.76). There was a significant difference between percentage of the hatched eggs and both treated and control groups. LC50 demonstrated resistance to Alb in treated groups; while it uncovered suspicion to drug resistance in Feb treated groups.

It was concluded that there was resistance to Alb and suspected resistance to Feb in sheep examined.

Chronic myeloid leukemia (CML) as a myeloproliferative disease is characterized by increased cellularity of bone marrow. Implementing the latest treatment protocols is currently accompanied by serious and life-threatening side effects. There are worldwide attempts to find new effective and potent therapeutic agents with minimal side effects on CML patients. This in vitro study was carried out to discover the potential antiproliferative and apoptotic effects of naturally produced prodigiosin (PDG) on K562 cells as an accepted model of CML.

The anti-proliferative effect of PDG was measured by MTT assay. To highlight the mechanism of cytotoxicity, the apoptotic cell death pathway was investigated by morphological and biochemical assessments. The dual acridine orange/ethidium bromide staining technique and western blotting method were applied to assess the mechanism of the potential apoptotic impact of PDG on K562 cells.

PDG-induced time- and concentration-dependent anti-proliferative effects were revealed with an estimated IC50 value of 54.06 μM. The highest cell viability reduction (60%) was recorded in cells, which were exposed to 100 μM concentration. Further assays demonstrated that in the dual acridine orange/ethidium bromide staining method the cell population in the late apoptosis phase was increased in a concentration-dependent manner, which was confirmed with remarkable DNA fragmentation.

We found that the PDG-induced apoptosis in K562 cells is mediated through the caspase-3 activation both in mRNA and protein levels. Our results suggest that PDG could be a potent compound for further pharmacokinetic and pharmacodynamics studies in the in vivo model of CML.

Acute toxicity assessment is the first priority in the determination of any related risk to the biologically unknown chemicals to human and animals. LD50 determination as an accepted model of acute toxicity assay in animal models for a new drug in clinical trials is one of the important requirements of the drug launching process. Prodigiosin is a substance extracted from Serratia marcescens and has antitumor and antifungal activities. Thus, in this study, acute toxicity of prodigiosin was determined using the lowest number of laboratory animals.

In this experimental study, different doses of prodigiosin were administered intraperitoneally in male mice. Twenty four hours after injection, alongside examining behavior of the animals receiving the prodigiosin, some organs including the heart, liver, kidney, spleen, intestine and lung were sampled, and after paraffin block preparation and microtome cutting, they were stained with hematoxylin-eosin and examined by light microscopy.

The results of this study indicated that LD50 for prodigiosin is 4500 mg / kg, when administered intraperitoneally and histopathological findings indicate very slight and minor damage to the liver, kidney and spleen, while no remarkable damage on other organs including the heart, lung and intestine was observed.

Based on the results of current study and estimated LD50 level, it is suggested that prodigiosin can be categorized as a safe compound with the least histopathological impact on the vital organs.

 Crocetin is a natural antioxidant that is found in the crocus flower and Gardenia jasminoides  (fruit). Previous studies have reported its anticancer activity both in vivo</em> and in vitro</em>. In addition, crocetin suppresses the growth and migration of human colorectal cancer cells, however, its mechanism of action remains to be elucidated. Therefore, the present study investigated the molecular mechanism of crocetin effect on colorectal cancer cells (HCT-116) in vitro.

 HCT-116 cells were treated with different concentrations (0, 200, 400, 600, and  800 μM) of crocetin for 24 h. The cell survival rate was measured by MTT assay. Cell migration capacity was evaluated using the wound healing assay. The expression levels of vascular endothelial growth factor (VEGF) and matrix metalloproteinase (MMP-9) was monitored by RT-PCR. Phosphorylation of focal adhesion kinase (FAK) and p38 mitogen-activated protein kinase (MAPK) was determined using western blot.

 The proliferation of HCT-116 was inhibited by crocetin at 800 μM (P</em> < 0.001). Crocetin prevented migration of HCT-116 cells (P</em> < 0.05) and suppressed VEGF and MMP-9 mRNA expression   (P</em> < 0.001) and increased phosphorylation of p38 (MAPK; P</em> < 0.001). However, no significant change in the phosphorylation of FAK was observed.

 These data suggested that crocetin-induced growth- and migration-suppressing effects on HCT-116 cells may partially depend on the regulation of the p38 (MAPK) signaling pathway. 

Molds and mycotoxins are contaminants of animal feed causing spoilage and clinical intoxication. Animal exposure to mycotoxins reflects diet composition with major differences occurring between animals kept predominantly of pastures, i.e. ruminants and horses, and those consuming formulated feed like pigs and poultry. Mixed feeds are composed of several ingredients, often sourced from different continents. Subsequently, practitioners may confront endemic diseases and signs of toxin exposure related to toxins imported accidentally with contaminated feed materials from other countries and continents. Mycotoxins comprise more than 300 to 400 different chemicals causing a variety of clinical symptoms. Mycotoxin exposure causes major economic losses due to reduced performance, impaired feed conversion and fertility, and increased susceptibility to environmental stress and infectious diseases.  In acute cases, clinical symptoms following mycotoxin ingestion are often non-specific, hindering an immediate diagnosis. Furthermore, most mold species produce more than one toxin, and feed commodities are regularly contaminated with various mold species resulting in complex mixtures of toxins in formulated feeds. The effects of these different toxins may be additive, depending on the level and time of exposure, and the intensity of the clinical symptoms based on age, health, and nutritional status of the exposed animal(s). Threshold levels of toxicity are difficult to define and discrepancies between analytical data and clinical symptoms are common in daily practice. This review aims to provide an overview of Aspergillus and Penicillium toxins that are frequently found in feed commodities and discusses their effects on animal health and productivity.

During the last two decades, anthelmintic drugs have been increasingly applied against gastrointestinal parasites of sheep in Iran.

For this purpose, drug resistance to albendazole (Alb) and fenbendazole (Feb) in gastrointestinal nematodes in sheep of Saqez multiplicity was assessed.

In in-vivo experiment, a total number of 90 sheep in three groups (30 sheep/group) with EPG≤150 were examined for nematode resistance to Alb and Feb. They were treated with Alb and Feb or untreated (as a control group).

There was significant difference between Alb and Feb treated groups and control group. The EPG in Alb, Feb and control groups was 59.8±1.93, 18.8±1.258 and 204.07±4.81, respectively. There was drug resistance against Alb in compassion with control group (R=71%). There was suspicion drug resistance for Feb in comparison with control group (R=90.66%).

From the results of the present study, it was concluded that there was absolute and suspected drug resistance against Alb and Feb in sheep of Saqez municipality, Iran, respectively.

In the present work, a miniaturized sample preparation method based on combination of dispersive solid phase extraction and temperature–induced homogenous liquid– liquid microextraction has been proposed for the extraction and preconcentration of some organophosphorus pesticides (parathion–methyl, triazophos, parathion, diazinon, and phoxim) from honey samples prior to their analysis by high performance liquid chromatography– ultraviolet detection.

In this method, initially the analytes were adsorbed onto a sorbent (C18) and then desorbed by the use of cyclohexyl amine as an eluent. In the next step, the eluent was mixed with water thermostated at 0 °C to obtain a homogenous solution. By increasing the temperature, the solubility of cyclohexyl amine in water was decreased and led to formation of dispersed fine droplets in the whole of solution. These droplets go up through the solution and collected on top of the solution. Finally, an aliquot of the organic phase was sucked in a microsyringe and injected into the separation system for analysis.

Under the optimum experimental conditions, limits of detection and quantification were calculated to be in the ranges of 0.90–1.75 and 3.0–5.8 ng g–1 in honey samples, respectively. Enrichment factors and extraction recoveries were in the ranges of 148–183 and 59–73%, respectively. The relative standard deviations varied from 2–4% and 4–5% for intra– (n = 6) and inter–day (n = 4) precisions, respectively.

The suggested approach was satisfactorily utilized to the analysis of 21 honey samples. The proposed miniaturized tandem sample pretreatment method enhanced the sensitivity of the instrumental analysis.

Tendon healing is substantially slow and often associated with suboptimal repair. Cell therapy is one of the promising methods to improve tendon repair. Blastema, a population of undifferentiated cells, represents characteristics of pluripotent mesenchymal stem cells and has the potentials to be used in regenerative medicine. The aim of this study was to investigate the use of blastema allotransplantation in rabbit tendon healing.

In this study, one rabbit was used as a blastema donor, and twenty-four rabbits were divided into control and treatment groups. Blastema cells were obtained from ear pinna upon punch hole injury in the donor rabbit. Under general anesthesia, a complete transverse tenotomy was performed on the midsubstance of deep digital flexor tendon followed by suture-repair. In the treatment group, 1 × 106 blastema cells suspended in buffer saline were injected intratendinously at the repair site, while the control group received only the buffer saline. Cast coaptation was maintained for two weeks. Eight weeks after the operation, tendons were harvested, and histopathological, biomechanical, and biochemical assays were performed on samples.

Mechanical testing showed a significant increase in ultimate load, energy absorption, stiffness, yield load, stress, and strain in blastema-treated tendons compared to controls. Also, higher hydroxyproline content and improved collagen alignment along with lower inflammatory cell infiltration and decreased angiogenesis were observed in blastema-treated tendons.

Increased levels of hydroxyproline and improved histopathological and biomechanical parameters in the treatment group suggest that blastema cells could be considered an adjunct to tendon repair in rabbits.