First molecular identification of Leishmania species in a new endemic area of cutaneous leishmaniasis in Lorestan,Iran

ObjectiveTo identify Leishmania using PCR.MethodsThis study was conducted from April 2009 to March 2011 in order to identify Leishmania species in a new endemic area of CL in Lorestan, Iran. Samples were taken from 62 patients that referred to the health centers in different cities of Lorestan province, the presence of Leishmania was confirmed using direct smear and then grown in NNN media and mass cultured in RPMI 1 640 medium supplemented with 10% heat-inactivated fetal bovine serum. DNA was extracted from cultured promastigotes and used in ITS-PCR.Results45(72.6%) samples out of 62 showed a band in the range of 485 bp and 17 (27.4%) with a band in the range of 626 bp which were similar to standard strains of Leishmania tropica(L. tropica) and Leishmania major(L. major), respectively. 50 (65.80%) of samples were collected from people with no history of travel in at least a year prior to the onset which shows that indigenous source of infection.ConclusionsSince the vector and reservoir of the two species are different, so precise and extensive control and prevention methods should be designed and carried out.     

Impact of Hepatitis C Virus Infection on Cognitive Function in Patients With Covert Hepatic Encephalopathy

Background:

Liver cirrhosis has a significant impact on patient’s quality of life and socioeconomic status and is associated with increased susceptibility to vehicle accidents and falls.

Objectives:

This study was conducted to evaluate cognitive function in patients with cirrhosis.

Patients and Methods:

All subjects, who underwent transient elastography (TE) between March 2014 and August 2014 in Shariati outpatient clinic, Tehran, Iran, were enrolled in the study. The diagnosis of cirrhosis was made based on clinical and laboratory findings and liver stiffness measurement (LSM) values > 14. Wechsler Memory Scale (WMS) and simple visual reaction time (RT) were the neuropsychiatric tests used to assess cognitive function, in all subjects.

Results:

A total of 37 cirrhotic patients and 37 matched controls were included. Patients with cirrhosis had significantly lower mean quotient (MQ) score, compared with controls (91 ± 18.2 vs. 114.6 ± 17.5, P <0.001). All of WMS subscales, except associate learning, showed statistically significant differences between the two groups. Reaction time was more prolonged in patients with cirrhosis, compared with controls. Cognitive function of hepatitis C virus (HCV) patients was significantly impaired, with respect to logical memory, visual reproduction and MQ score, compared to other underlying causes of cirrhosis (P = 0.01, P = 0.04, and P = 0.01, respectively)

Conclusions:

Our results suggested that neuropsychiatric and cognitive problems must be considered in the management of covert hepatic encephalopathy (HE) patients, especially in patients with HCV infection. The WMS and simple visual RT tests are useful methods for the early diagnosis of covert HE.     

Mathematical beta function formulation for maxillary arch form prediction in normal occlusion population

The aim of this study was to assess the dental arch curvature in subjects with normal occlusion in an Iranian population and propose a beta function formula to predict maxillary arch form using the mandibular intermolar widths (IMW) and intermolar depths (IMD). The materials used were study casts of 54 adolescents with normal occlusion and mean age of 14.1 years (25 males, 29 females, age range 12–16 years). Curve-fitting analyses were carried out and the curves passing through the facial-axis point of the canines, premolars, first molars, and the incisal edges of the anterior teeth were studied using a 3D laser scanner. Using the measured IMW and IMD of the dental arches at the maxillary and mandibular first molar region, a beta function formula proposed for predicting maxillary arch form. The accuracy of the proposed formula was assessed on 10 randomly selected dental casts. The mean (SD) of the maxillary and mandibular IMW and IMD were 57.92 (4.75), 54.19 (5.31), and 31.59 (2.90) and 28.10 (2.59) mm, respectively. There was no gender dimorphism (P > 0.05) for both variables (IMW, IMD). There was a strong positive association (n = 10, Pearson r = 0.98, P < 0.05) between the measured (actual) maxillary arch length and proposed arch length derived from generated formula. The goodness of fit (whole arch) for the proposed beta function formula, using adjusted r square measure and root mean square in 10 patients averaged 0.97 and 1.49 mm, respectively. The corresponding figures for the maxillary anterior arch (canine to canine) were 0.90 and 0.92 mm, respectively. The proposed beta function formula used for predicting maxillary arch form based on two mandibular measures (IMW, IMD) was found to have a high accuracy for maxillary arch prediction in the Iranian population and may be used as a guide to fabricate customized arch wires or as an aid in maxillary reconstructive surgery.     

Prospects for antimicrobial peptide-based immunotherapy approaches in Leishmania control

Introduction: Leishmaniasis is one of the neglected tropical diseases and is highly endemic in many countries. Currently, there is no adequate human vaccine and treatment to control the disease.

Areas covered: As a result of the failure of chemotherapy and toxicity, it is necessary to find another approach for the treatment of leishmaniasis. Recently, antimicrobial peptides (AMPs), originating from natural resources, have attracted much attention for their use as a new antibiotics for many infectious and noninfectious diseases. Natural AMPs are named interchangeably as host defense peptides. They are naturally active in the innate immune system as a primary defense mechanism in most species all over the world. Several AMPs have been tested in in vitro and in vivo experiments against leishmaniasis.

Expert commentary: Most AMPs require proper conformation to be active. Leishmania (L.) tarentolae as a nonpathogenic strain, is an effective tool not only for vaccine development but also for therapy. Recombinant L. tarentolae expressing selective or combined AMPs is a suggestive approach for leishmaniasis or any other infectious disease treatment.     

Effects of prolonged fasting on fatigue and quality of life in patients with multiple sclerosis

Fasting is one of the recommended worships of several great religions in the world. During the month of Ramadan, circadian rhythm and pattern of eating changes result in physiological, biochemical and hormonal changes in the body. Many Muslims with medical conditions ask their physicians about the feasibility and safety of fasting during Ramadan. In this study, we aim to assess the effect of Ramadan fasting on the quality of life and fatigue in multiple sclerosis (MS) patients. Relapsing-remitting MS (RRMS) patients according to McDonald’s criteria who had mild disability (EDSS score ≤3) were included in this study. Fatigue and quality of life were were assessed using the validated Persian versions of modified fatigue impact scale (MFIS) and multiple sclerosis quality of life-54 (MSQOL-54) questionnaires, respectively. 218 patients (150 females and 68 males) were enrolled in our study. There was no statistically significant difference between the mean total score of MSIF before and after fasting (25.50 ± 13.81 versus 26.94 ± 16.65; p = 0.58). The mean physical health and mental health composites of quality of life increased significantly after fasting (p = 0.008 and p = 0.003 respectively). Despite the observed lack of favorable effects on fatigue, our results showed increased quality of life of MS patients once Ramadan has ended. Whether this is specifically related to Ramadan-related fasting deserves further testing in appropriately designed larger prospective clinical studies.     

Recombinant Leishmania tarentolae expressing the A2 virulence gene as a novel candidate vaccine against visceral leishmaniasis

Visceral leishmaniasis is the most severe form of leishmaniasis. To date, there is no effective vaccine against this disease. Many antigens have been examined so far as protein- or DNA-based vaccines, but none of them conferred complete long-term protection. The use of live attenuated vaccines has recently emerged as a promising vaccination strategy. In this study, we stably expressed the Leishmania donovani A2 antigen in Leishmania tarentolae, a non-pathogenic member of the genus Leishmania, and evaluated its protective efficacy as a live vaccine against L. infantum challenge. Our results show that a single intraperitoneal administration of the A2-recombinant L. tarentolae strain protects BALB/c mice against L. infantum challenge and that protective immunity is associated with high levels of IFN-γ production prior and after challenge. This is accompanied by reduced levels of IL-5 production after challenge, leading to a potent Th1 immune response. In contrast, intravenous injection elicited a Th2 type response, characterized by higher levels of IL-5 and high humoral immune response, resulting in a less efficient protection. All together, these results indicate the promise of A2-expressing L. tarentolae as a safe live vaccine against visceral leishmaniasis.     

Compatibility studies of acyclovir and lactose in physical mixtures and commercial tablets

This study documents drug-excipient incompatibility studies of acyclovir in physical mixtures with lactose and in different tablet brands. Differential scanning calorimetry (DSC) was initially used to assess compatibility of mixtures. The Fourier-transform infrared (FTIR) spectrum was also compared with the spectra of pure drug and excipient. Although DSC results indicated incompatibility with lactose, FTIR spectra were mostly unmodified due to overlapping peaks. Samples of isothermally stressed physical mixture were stored at 95 °C for 24 h. The residual drug was monitored using a validated high-performance liquid chromatography (HPLC) assay and data fitting to solid-state kinetic models was performed. The drug loss kinetics followed a diffusion model. The aqueous mixture of drug and excipient was heated in order to prepare an adduct mixture. HPLC analysis revealed one extra peak that was fractionated and subsequently injected into the liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) system. The MRM (Multiple Reaction Monitoring) chromatograms characterized the peak with molecular mass corresponding to an acyclovir-lactose Maillard reaction product. The presence of lactose in commercial tablets was checked using a new TLC method. Overall, the incompatibility of acyclovir with lactose was successfully evaluated using a combination of thermal methods and LC-MS/MS.     

Interaction of CpG-oligodeoxynucleotides with Toll like receptor 9 induces apoptosis and modulates metaloproteinase-2 activity in human intestinal epithelium

Recent reports have indicated different effects of immunostimulatory sequences containing CpG-Oligodeoxynucleotides (ODN) on various immune cells. However, the exact role of CpG-ODN in the human gut is unclear. In the present study, we assessed potential effects of CpG-ODN on non lymphoid cell (intestinal epithelial cell line HT-29) on a dose-response and time-course basis. Intestinal epithelial cell line HT-29 was treated with CpG-ODN (CpG 2006) and lipopolysaccharide (LPS) at 5, 10, 25, 50 microg/ ml and 1, 5, 10 microg/ ml concentrations, respectively. Following treatments, dose- response and time-course cytotoxicity using a colorimetric method, Metaloproteinase-2 (MMP-2) activity (using gelatin zymography) and apoptosis (using annexin-v flowcytometry method) assays were performed. Chloroquine treatment was also used for its inhibitory effect on endosomal acidification process to verify specific CpG-ODN and Toll Like Receptor 9 (TLR9) interactions. Cytotoxicity analysis of CpG-ODN showed that CpG-ODN increased significantly the proliferation of CpG-ODN treated cells, as compared to untreated cells, at concentrations of 10-25 microg/ml (p < 0.05). Overall MMP-2 activity analysis showed significant differences between treated and untreated cells. However, minimal changes were observed when MMP-2 activity was assessed per cell. Moreover, CpG-ODN treated cells demonstrated an increasing apoptosis rate of 0.8 %, 6.46 % and 14.21% at concentrations of 5, 10, 25 microg/ml, respectively. Collectively, our data indicated that intestinal epithelial cell line HT-29 is highly responsive to CpG effect in vitro and exhibits modified activities. The direct CpG-ODN and TLR-9 interactions in HT-29 cells could provide new approaches in malignant tumor therapeutic strategies.