Evaluation of antioxidant potential of Heliotropium bacciferum Forssk extract and wound healing activity of its topical formulation in rat.

Wound healing is a dynamic process that occurs in the tissue under the skin. During this process, oxidative stress biomarkers are excessively produced, which finally lead to inflammation and cellular damage. In this study, efforts have been made to evaluate the antioxidant effect and wound healing activity topical formulation containing Heliotropium bacciferum Forssk extract. The in vitro antioxidant properties were evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activities and the ferric reducing antioxidant power (FRAP) assay. The herbal ointments (2.5% w/w and 5% w/w) were prepared from the hydroalcoholic extract of H. bacciferum Forssk and administrated on the induced wounds in Wistar rats. The chromatic assay, percentage of wound contraction, and histopathological studies were used for evaluating the wound healing activity. For the evaluation of reactive oxygen species (ROS), catalase (CAT) activity, superoxide dismutase (SOD), and glutathione (GSH) levels were examined. The DPPH method showed tremendous radical scavenging activities at the corresponding concentrations with EC50 value of 80µg/mL. Topical application of the ointment (5% w/w) showed the highest wound contraction in comparison to the positive control (treated with CICALFATE™) and the control group (treated with normal saline). Similarly, the histological study of the group treated with the extract ointment (5% w/w) showed full collagen tissue deposition with a complete epidermal regeneration. The results of the assessment of GSH levels as well as CAT and SOD activities in the treated group (5% w/w) confirmed the scavenging property of the extract ointment. Our findings indicated the proper wound healing impact of the topical formulation of H. bacciferum Forssk due to its notable antioxidant capacity.

Effect of hydroxychloroquine on COVID-19 prevention in cancer patients undergoing treatment: study protocol for a randomized controlled trial.

OBJECTIVES: In this study, we will investigate the effect of hydroxychloroquine on the prevention of novel coronavirus disease (COVID-19) in cancer patients being treated. TRIAL DESIGN: This is a two-arm, parallel-group, triple-blind, phase 2-3 randomized controlled trial. PARTICIPANTS: All patients over the age of 15 years from 5 types of cancer will be included in the study. Patients with acute lymphoid and myeloid leukemias in the first line treated with curative intent, patients with high-grade non-Hodgkin's lymphoma treated with leukemia regimens, and patients with non-metastatic breast and colon cancer in the first line of treatment will enter the study. INTERVENTION AND COMPARATOR: Patients are randomly assigned to two groups: one being given hydroxychloroquine and the other is given placebo. During 2 months of treatment, the two groups will be treated with hydroxychloroquine every other day with a single 200-mg tablet (Amin® Pharmaceutical Company, Isfahan, Iran) or placebo (identical in terms of shape, color, and smell). Patients will be monitored for COVID-19 symptoms during follow-up period. If any COVID-19-related signs or symptoms occur, they will be examined, thoroughly, investigated with a high resolution computerize tomography (CT) scan of the lungs and nasopharyngeal swab assessed by RT-PCR for SARS-CoV-2 virus. This study will be performed in five centers affiliated to Mashhad University of Medical Sciences, Mashhad, Iran. MAIN OUTCOMES: The primary end point of this study is to investigate the incidence of COVID-19 in patients being treated for their cancer and receiving prophylactic Hydroxychloroquine. RANDOMIZATION: Randomization will be performed using random permuted blocks. By using online website ( www.randomization.com ), the randomization sequence will be produced by quadruple blocks. The allocation ratio in intervention and control groups is 1:1. BLINDING (MASKING): Participants and caregivers do not know whether the patient is in the intervention or the control group. Those assessing the outcomes and data analyzer are also blinded to group assignment. SAMPLE SIZE: The calculated total sample size is 60 patients, with 30 patients in each group.

No longer a historical ailment: two cases of childhood scurvy with recommendations for bone health providers.

PURPOSE: Scurvy, due to vitamin C deficiency, is commonly referenced as a "forgotten" or "historical" disease. A growing number of case reports challenge this notion. Bone health providers are often consulted early in the presentation of scurvy to evaluate musculoskeletal complaints resulting from impaired collagen production and disrupted endochondral bone formation. In this report, we describe two cases of childhood scurvy. Our objective is to summarize the key features of scurvy for bone health providers, with the goal of raising awareness and facilitating diagnosis in future cases. CASE DESCRIPTIONS: Case one occurred in a 12-year-old non-verbal, non-ambulatory female on a ketogenic diet for refractory epilepsy. Clinical findings included hemarthrosis, transfusion dependent anemia, elevated inflammatory markers, and epiphysiolysis. Magnetic resonance imaging (MRI) revealed multi-focal bone marrow signal abnormalities and physeal irregularities. Case two occurred in a typically developing 5-year-old male presenting with limp and knee pain. Symptoms progressed despite casting and immobilization. Mild anemia, elevated inflammatory markers, and multi-focal marrow and physeal MRI abnormalities were identified. Subsequent dietary history revealed total absence of fruit or vegetable consumption. The diagnosis of scurvy was confirmed in both cases by undetectable plasma vitamin C concentrations. Treatment with vitamin C led to rapid clinical improvement. CONCLUSION: Scurvy can no longer be considered a historical diagnosis and should not be forgotten when evaluating children with musculoskeletal ailments. Early recognition of the signs, symptoms, and imaging findings of scurvy can reduce the clinical burden of this disease with the timely initiation of vitamin C therapy.

Anisakis pegreffii impacts differentiation and function of human dendritic cells.

Human dendritic cells (DCs) show remarkable phenotypic changes when matured in the presence of helminth-derived products. These modifications frequently elicited a polarization towards Th2 cells and regulatory T cells thus contributing to immunological tolerance against these pathogens. In this study, the interaction between DCs and larvae of the zoonotic anisakid nematode Anisakis pegreffii was investigated. A. pegreffii larvae were collected from fish hosts, and monocyte-derived DCs were cocultured in the presence of the live larvae (L) or its crude extracts (CE). In both experimental conditions, A. pegreffii impacted DC viability, hampered DC maturation by reducing the expression of molecules involved in antigen presentation and migration (ie HLA-DR, CD86, CD83 and CCR7), increased the phagosomal radical oxygen species (ROS) levels and modulated the phosphorylation of ERK1,2 pathway. These biological changes were accompanied by the impairment of DCs to activate a T-cell-mediated IFNγ. Interestingly, live larvae appeared to differently modulate DC secretion of cytokines and chemokines as compared to CE. These results demonstrate, for the first time, the immunomodulatory role of A. pegreffii on DCs biology and functions. In addition, they suggest a dynamic contribution of DCs to the induction and maintenance of the inflammatory response against A. pegreffii.

Evaluation of PCR-reverse line blot hybridization assay for simultaneous identification of medically important saprophytic fungi.

BACKGROUND: In immunocompromised patients suffering from invasive fungal infections, rapid identification of fungal species is important since the appropriate treatment is usually related to the responsible species. We describe here, an assay based on combination of PCR and reverse line blot hybridization (PCR/RLB) for differentiation causative agent of fungal infections. MATERIALS AND METHODS: We performed PCR/RLB assay on 10 reference strains, which include Aspergillus species (A. fumigatus, A. flavus, A. niger, A. terreus, and A. clavatus), Mucor circnelloides, Rhizopus oryzae, Alternaria alternata, Cladosporium herbarum, and Fusarium solani. Besides, twenty-two clinical specimens from patients with proven fungal infections were analyzed for the identification of species. The obtained results were then compared with the results of culture and sequence analysis. RESULTS: The fungal species-specific oligonucleotide probes were able to distinguish between all species represented in this study with the exception of cross-reactivity between A. niger and A. fumigatus species. Two specimens, which were represented as mixed fungi in culture, were identified properly by this method. Results of the RLB assay were concordant with the culture and ITS sequencing results. CONCLUSION: Our result demonstrate that the RLB assay potentially is suitable for rapid and simultaneous identification of variety fungal pathogens directly from culture as well as from clinical specimens.

Pharmacokinetic Comparison of Omeprazole Granule and Suspension Forms in Children: A Randomized, Parallel Pilot Trial.

Although, omeprazole is widely used for treatment of gastric acid-mediated disorders. However, its pharmacokinetic and chemical instability does not allow simple aqueous dosage form formulation synthesis for therapy of, especially child, these patients. The aim of this study was at first preparation of suspension dosage form omeprazole and second to compare the blood levels of 2 oral formulations/dosage forms of suspension & granule by high performance liquid chromatography (HPLC). The omeprazole suspension was prepared; in this regard omeprazole powder was added to 8.4% sodium bicarbonate to make final concentration 2 mg/ml omeprazole. After that a randomized, parallel pilot trial study was performed in 34 pediatric patients with acid peptic disorder who considered usage omeprazole. Selected patients were received suspension and granule, respectively. After oral administration, blood samples were collected and analyzed for omeprazole levels using validated HPLC method. The mean omeprazole blood concentration before usage the next dose, (trough level) were 0.12±0.08 µg/ml and 0.18±0.15 µg/ml for granule and suspension groups, respectively and mean blood level after dosing (C2 peak level) were 0.68±0.61 µg/ml and 0.86±0.76 µg/ml for granule and suspension groups, respectively. No significant changes were observed in comparison 2 dosage forms 2 h before (P=0.52) and after (P=0.56) the last dose. These results demonstrate that omeprazole suspension is a suitable substitute for granule in pediatrics.

A structural model of the anaphase promoting complex co-activator (Cdh1) and in silico design of inhibitory compounds.

Anaphase promoting complex (APC) controls cell cycle and chromosome segregation. The APC activation occurs after binding of co-activators, cdh1 and cdc20. Cdh1 plays a role in cancer pathogenesis and is known as a potential drug target. The main aim of this study was prediction of 3D structure of cdh1 and designing the inhibitory compounds based on the structural model. First, 3D structure of cdh1 was predicted by means of homology modelling and molecular dynamics tools, MODELLER and Gromacs package, respectively. Then, inhibitory compounds were designed using virtual screening and molecular docking by means AutoDock package. The overall structure of cdh1 is propeller like and each DW40 repeat contains four anti-parallel beta-sheets. Moreover, binding pocket of the inhibitory compounds was determined. The results might be helpful in finding a suitable cdh1 inhibitor for the treatment of cancer.

Dietary intake of patients with angiographically defined coronary artery disease and that of healthy controls in Iran.

OBJECTIVE: The aim of this study was to investigate the relationship between dietary intake and biomarkers of cardiovascular risk in individuals with and without angiographically defined coronary artery disease. SUBJECTS/METHODS: Coronary angiography was undertaken in 445 individuals who were divided into those with significant disease (>50% occlusion) (Angio+ (n=273)) and those with <50% coronary artery occlusion (Angio- (n=172)). Apparently healthy, non-symptomatic individuals (n=443) were considered as the control group. Dietary intake was assessed using a 24-h dietary recall method and dietary analysis was performed using Diet Plan 6 software. RESULTS: Concentrations of starch, saturated fatty acids, polyunsaturated fatty acids, magnesium, iron and copper in the control group were less than those in the other groups (P<0.05), but after adjusting for total energy intake these differences were no longer apparent. The mean intake of protein, cholesterol, phosphorus, zinc, zinc/copper ratio, selenium, iodine, carotene, vitamin E, niacin, pantothene and pyridoxine was less in the control group compared with the other two groups (P<0.05), and the mean of sugar, fiber, transfatty acids, manganese, folate and vitamin C was higher in the control group than in other groups (P<0.05). Lipid profile values between the three groups did not differ significantly. CONCLUSIONS: These results indicate that the amount of intake of various nutrients can be considered as an independent risk factor for CAD. Further research on the relationship between CAD and nutrient intake, especially intake of essential micronutrients, is needed.

Aberrant regulation of the integrin very late antigen-4 in systemic lupus erythematosus.

Integrin very late antigen-4 (VLA4) is induced during inflammation and can regulate monocyte migration. It has been implicated in atherogenesis, a significant concern in systemic lupus erythematosus (SLE). The aim of this study was to define VLA4 expression in SLE monocytes. Flow cytometry, reverse transcription polymerase chain reaction, Western blotting, and immunohistochemistry staining with confocal microscopy were used to evaluate VLA4 expression in SLE patients and controls. We found elevated expression of VLA4 in SLE patients with significantly increased VLA4 staining intracellularly compared to control. Exposure of control monocytes to SLE sera or immune complexes led to increased intracellular expression, and immune complexes were capable of driving redistribution of surface VLA4 to the cytoplasm. Therefore, VLA4 was found to be subject to complex regulation with SLE sera driving both RNA expression and redistribution of protein. Stimulation of SLE monocytes with a VLA4 ligand induced significant TNFα expression, confirming a functional effect. This behavior may contribute to increased atherosclerosis and monocyte infiltrates in end organs.

Past, present and future strategies of immunotherapy in gynecological malignancies.

Recently, the combining of different drugs has greatly improved response and survival rates in gynecological malignancies. Results are however far from being satisfactory. Treatments used in case of advanced or recurrent disease offer limited results in terms of long-term responses. The urgent need for new and more effective treatments has prompted researchers to investigate and propose new therapeutic strategies. One of the most interesting approaches that are being explored is constituted by immunotherapy. Currently, immunotherapeutic strategies include vaccination with peptide, viral vectors, carbohydrates and antiidiotypic antibodies. In addition, cell based immunotherapy has been adopted in vitro activated lymphocytes and dendritic cells. Most experience has been acquired in ovarian cancer and cervical cancer. Little has been investigated in endometrial and rare gynecologic neoplasms.The clinical experiences and results achieved with immunotherapy in this setting of patients have been reviewed and the future avenues that are currently being explored have also been discussed.

Spry1 and spry2 are essential for development of the temporomandibular joint.

The temporomandibular joint (TMJ) is a specialized synovial joint essential for the function of the mammalian jaw. The main components of the TMJ are the mandibular condyle, the glenoid fossa of the temporal bone, and a fibrocartilagenous disc interposed between them. The genetic program for the development of the TMJ remains poorly understood. Here we show the crucial role of sprouty (Spry) genes in TMJ development. Sprouty genes encode intracellular inhibitors of receptor tyrosine kinase (RTK) signaling pathways, including those triggered by fibroblast growth factors (Fgfs). Using in situ hybridization, we show that Spry1 and Spry2 are highly expressed in muscles attached to the TMJ, including the lateral pterygoid and temporalis muscles. The combined inactivation of Spry1 and Spry2 results in overgrowth of these muscles, which disrupts normal development of the glenoid fossa. Remarkably, condyle and disc formation are not affected in these mutants, demonstrating that the glenoid fossa is not required for development of these structures. Our findings demonstrate the importance of regulated RTK signaling during TMJ development and suggest multiple skeletal origins for the fossa. Notably, our work provides the evidence that the TMJ condyle and disc develop independently of the mandibular fossa.

Association of interleukin-15 single nucleotide polymorphisms with resistance to brucellosis among Iranian patients.

Interleukin (IL)-15, a Th1-related cytokine, triggers inflammatory cells' recruitment and increases the expression of interferon gamma (IFN-γ), which is an important cytokine in the immunity against brucellosis. Different single-nucleotide polymorphisms (SNPs) have been observed in the IL-15 gene, so this study aimed to investigate the probable association between these SNPs and susceptibility to brucellosis among Iranian patients. A total of 190 patients with brucellosis and 83 healthy milk farmers who consumed contaminated raw milk and dairy products from animals involved with brucellosis were included in this study. All the patients and the controls were genotyped for four IL-15 polymorphisms at positions 267, 367, 13687 and 14035 using polymerase chain reaction-restriction fragment length polymorphisms. The 267C and 13687A alleles, haplotypes CGCT and CAAA and the 267CC and 13687AA genotypes were significantly more frequent in the controls than in the patients (P = 0.014, 0.03, 0.006, 0.024, 0.026 and 0.01, respectively), so the variation in the IL-15 gene may be one of the factors affecting the resistance to brucellosis. In contrast, the frequency of haplotypes CGCA and TACT was significantly higher in patients compared with controls (P = 0.015 and 0.007, respectively), and interestingly the last one was observed only in the patients; therefore, it may serve as a predictive factor for brucellosis. In conclusion, it could be suggested that IL-15 genetic variants can affect resistance or susceptibility to human brucellosis among Iranian patients.

Antioxidant status in patients with lichen planus.

BACKGROUND: Lichen planus (LP) is an autoimmune inflammatory disease of the mucocutaneous tissue, whose exact pathological course is not yet understood. Many studies have implicated the role of reactive oxygen species (ROS) and the protective role of antioxidants in several autoimmune skin disorders. In this study, serum levels of antioxidants in patients with LP were determined and compared with those of healthy controls. METHODS: In total, 30 patients with LP (mean ± SD age 41.63 ± 13.03), who had never received treatment for their disease, were enrolled; 30 healthy people (aged 41.17 ± 13.24) were recruited as the control group. Serum levels of glutathione peroxidase (GPX), vitamin C, selenium, bilirubin and uric acid were determined. RESULTS: The mean plasma level of vitamin C was significantly lower (P < 0.001) in patients compared with controls. A significant positive correlation was found between selenium and GPX in both patients (Spearman ρ = 0.99, P < 0.001) and controls (ρ = 0.10, P < 0.001). CONCLUSIONS: Lower serum levels of vitamin C in patients with LP indicates that free radicals and the resulting oxidative damage may be important in the pathogenesis of LP lesions.

Performance of antigen B isolated from different hosts and cyst locations in diagnosis of cystic echinococcosis.

BACKGROUND: The aim of this study was to assess the performance of Antigen B (AgB) isolated from different Echinococcusgranulosus intermediate hosts and from different cyst locations for immunodiagnosis of human cystic echinococcosis (CE). METHODS: Hydatid cyst fluids were collected from lung and liver cysts of sheep, liver cysts of goats, lung cysts of camels and cattle, and liver cysts of human. AgB was purified from each of these hydatid cysts fluids. Serum samples obtained from 47 pathologically confirmed cases of CE along with 30 sera samples from non-CE patients and 40 sera from healthy individuals were tested by ELISA using AgB prepared from different hosts or cyst locations. RESULTS: The highest sensitivity (97.8%) for diagnosis of CE was seen with AgB prepared from human liver cysts. This maximal sensitivity was followed by AgB isolated from those of sheep liver and lung cysts. The least sensitivity was found with AgB prepared from bovine lung cysts. The highest specificities (97.1%) were observed with AgB isolated from human liver cysts followed by those of sheep and goat liver cysts while the lowest specificity was seen with AgB isolated from bovine lung cysts. In view of the specificities and sensitivities of the different AgB, the best validity was found for AgB prepared from human liver cysts while the least validity was found with AgB prepared from bovine lung cysts. CONCLUSION: For any AgB-based tests, obtaining of the antigen from one of these sources will significantly increase the diagnostic sensitivity and specificity of the assay.

Lipoid Proteinosis in two Iranian Sisters: A Case Report and Review of Literature.

Lipoid proteinosis is a rare autosomal recessive disorder which may be seen within a family very occasionally. Herein, we report lipoid proteinosis in two sisters characterized by verrucous lesions and hoarseness of voice, dysphagia and multiple beaded papules along the margins of their eyelids, fissured lips and thick ferenulum.

Habitat characterization and mapping of Anopheles maculatus (Theobald) mosquito larvae in malaria endemic areas in Kuala Lipis, Pahang, Malaysia.

In Peninsular Malaysia, a large proportion of malaria cases occur in the central mountainous and forested parts of the country. As part of a study to assess remote sensing data as a tool for vector mapping, we conducted entomological surveys to determine the type of mosquitoes, their characteristics and the abundance of habitats of the vector Anopheles maculatus in malaria endemic areas in Pos Senderot. An. maculatus mosquitoes were collected from 49 breeding sites in Pos Senderot. An. maculatus preferred to breed in water pockets formed on the bank of rivers and waterfalls. The most common larval habitats were shallow pools 5.0-15.0 cm deep with clear water, mud substrate and plants or floatage. The mosquito also preferred open or partially shaded habitats. Breeding habitats were generally located at 100-400 m from the nearest human settlement. Changes in breeding characteristics were also observed. Instead of breeding in slow flowing streams, most larvae bred in small water pockets along the river margin.

Immunogenicity of allo-vesicle carrying ERBB2 tumor antigen for dendritic cell-based anti-tumor immunotherapy.

Dendritic cells (DCs) are able to orchestrate innate and acquired immunity and can activate and sustain a long-lasting anti-tumor immune response in vivo when used as anti-tumor cell therapy. The selection of the antigen and the choice of its formulation are key points in designing anti-cancer DC-based vaccines. Cell released vesicles/exosomes have been shown to transfer antigens, HLAI/peptide complexes and co-stimulatory molecules to recipient cells. In this study we describe the generation of an allogenic microvesicle cell factory in which the expression of a specific tumor antigen was combined to the expression of co-stimulatory and allogeneic molecules. The DG75 lymphoblastoid cell line was selected as microvesicle producer and transfected with ErbB2, as tumor antigen prototype. The shed microvesicles transferred antigenic components to recipient DCs, increasing their immunogenicity. DC pulsing resulted in cross-presentation of ErbB2 both in HLAI and HLAII compartments, and ErbB2-specific CD8+ T cells from cancer patients were activated by DCs pulsed with vesicle-bound ErbB2. The microvesicle cell factory proposed may represent a source of cell free immunogen to be used for DC-based cancer therapy.

Dietary betaine affect duodenal histology of broilers challenged with a mixed coccidial infection.

The purpose of this research was to investigate effect of dietary betaine on intestinal morphology after an experimental coccidiosis. Hence a total of 189 male and female broiler chicks were randomly assigned to 9 floor cages. Chicks were fed a basal diet supplemented with 0, 0.6 or 1.2 g kg(-1) betaine. All birds were inoculated orally with Eimeria oocysts on day 28. Duodenal morphology parameters and lesions were scored by microscopic observation on intestine samples which were taken at day 42 of age. Adding 1.2 g kg(-1) betaine to diet diminished intestinal lesions (p < 0.05). Dietary supplementation with 0.6 or 1.2 g kg(-1) betaine significantly (p < 0.01) increased intraepithelial lymphocytes as well. Level of additive betaine had no effect on the ratio of villus height/crypt depth or villus surface area. Lamina propria of duodenum became thicker in the intestine of chickens which received more supplemental betaine via their diet. In conclusion, since the number of intraepithelial lymphocytes and thickness of lamina propria represent the condition of gut immune response, it seems that dietary betaine may immunomodulate the gastrointestinal tract of broilers. In addition, betaine effect on villus morphology measured later in life differed from what had been measured already earlier in life of the chicks.

[Attitudes and knowledge about smoking among Moroccan physicians looking after patients with tuberculosis]. / Attitudes et connaissances des pneumo-phtisiologues marocains vis-à-vis du tabagisme chez leurs patients tuberculeux.

Cigarette smoking is known to be a risk factor for the development of pulmonary tuberculosis; the management of the disease offers a good opportunity for smoking prevention. The aim of this study was to evaluate attitudes and knowledge of Moroccan respiratory physicians towards smoking in the management of patients with tuberculosis and the feasibility of integrating anti-smoking interventions into the national anti-TB program (PNLAT). A questionnaire was administered using a phone interview to Moroccan respiratory physicians practicing at the Tuberculosis and Respiratory Diagnostic Centre (CDTMR). 75 respiratory physicians answered the questionnaire representing 83.3% of all those approached. The rate of smoking in medical doctors was 10.7%. 66.7% of those interviewed considered that smoking increases the incidence of tuberculosis while 96% believed that smoking would worsen the pathology. More than 84% inquired about the smoking habits of their patients. Only 5.3% believed that they were well trained to help smokers to stop. 78.7% recognized the desirability of integrating a systematic smoking cessation program into the national anti-TB strategy. The rate of smoking among patients with TB is considered high by Moroccan doctors caring for the condition. However, they do not feel that they are sufficiently trained in smoking cessation techniques to help their patients. Therefore, education on smoking cessation needs to be integrated into the national anti-TB strategy.