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.

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.

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.

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.

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.

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.

[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.

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.

[An unusual cause of hemoptysis: a leech]. / Une cause inhabituelle d'hémoptysie: la sangsue.

We report an exceptional case of hemoptysis observed in a 38-year-old woman with an uneventful past history. Bronchial fibroscopy demonstrated the presence of a leech attached to the laryngeal orifice. This condition is common in endemic areas but is underdiagnosed. The association of hemoptysis, voice alterations, and difficult respiration are suggestive signs. Extraction should be achieved rapidly to avoid death due to acute respiratory failure.

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.

Human antibodies against the polymorphic epithelial mucin in ovarian cancer patients recognise a novel sequence in the tandem repeat region.

The humoral immune response to the polymorphic epithelial mucin (PEM) was studied by characterising the reactivity of human antibodies generated by EBV-immortalised B-cells from tumour-draining lymph nodes of ovarian cancer patients. All the human antibodies, selected in ELISA for their reactivity to the protein tandem core repeat sequence, reacted with PEM-expressing tumour cells. Aberrant glycosylation of the peptide core of the PEM molecule in cancer cells leads to the exposure of peptide epitopes that can be considered tumour specific. The epitope mapping of six human antibodies revealed that only one of them contained the PDTR sequence, shown to be the immunodominant epitope in the mouse. Four of the six human antibodies recognised a novel common immunogenic sequence (APPAH) in the tandem repeats. The binding of these human antibodies did not appear to be modulated by the length of the carbohydrate side chains, as shown by O-glycosylation inhibition studies. These results indicate that distinct sequences within the tandem repeat of PEM are target for a humoral immune response in humans. The presence of antibodies directed against different epitopes within the same antigenic region may modulate the antigen presentation process and the ongoing immune response. This data may help in clarifying the mechanisms of the immune response to PEM in cancer patients for the development of PEM-based immunotherapy.

Modulation of blood circulating immune cells by radiofrequency tumor ablation.

Tumor ablation by radiofrequency (RFA) is an appealing therapeutical strategy for the treatment of liver tumors (hepatocarcinoma and metastatic lesions) to be used as valid alternative to the surgical resection that often is appropriate and feasible in only a minority of patients. RFA induces the localised and controlled disruption of the tumor by heating the tissue causing its coagulative necrosis. Such therapy results as a pathogenic "noxa" for the body, inducing a strong inflammatory response. We wanted to ascertain whether the inflammatory response induced by RFA was similar in patients with hepatocarcinoma and in patients with liver metastasis. We considered body temperature, leucocyte counts at different time points as inflammatory parameters. We observed that RFA treatment produced the inflammatory systemic effects as expected (fever, increase of neutrophils) only in the patients with liver metastasis, while no such effect could be seen in the HCC patients. On the other hand the circulating monocytes increased after RFA in both groups of patients. These preliminary results suggest that RFA tratment can exert different effects on the immune system depending the etiopathogenesis of the treated neoplastic liver lesions.

A study of the sympathetic skin response and sensory nerve action potential after median and ulnar nerve repair.

The purpose of this study was to compare SSR with sensory nerve action potential (SNAP) responses in regeneration of injured peripheral nerves after nerve repair. We studied 10 male patients with a mean age of 26.7 years. All the patients had complete laceration of median or ulnar nerves. The patients were followed up at least for six months. SSR and SNAP assessment were performed every one to two months. Normal hands were used as controls. SSR was positive after 15.8 +/- 9.4 weeks (mean +/- 2 SD) and SNAP after 27.8 +/- 12.9 weeks (mean +/- 2 SD). The difference was statistically significant (P value < 0.001). This can be due to more rapid growth of sympathetic unmyelinated fibers relative to sensory myelinated fibers. This study also shows that recovery of the sudomotor activity following nerve repair is satisfactory in general and SSR can be used as a useful and sensitive method in the evaluation of sudomotor nerve regeneration.

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.

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.