Correlation between clinical severity score and the lower esophageal sphincter relaxation pressure in idiopathic achalasia

OBJECTIVES: Achalasia is an esophageal motor disorder characterized by aperistalsis and incomplete relaxation of the lower esophageal sphincter (LES). The meaningful correlation between LES relaxation pressure and the severity of clinical symptoms is uncertain. The aim of this study was to elucidate the correlation between the clinical scoring and the LES relaxation pressure. METHODS: Over a 4-yr period from 1997 to 2001, all newly diagnosed patients with idiopathic achalasia were consecutively enrolled in a study. Diagnosis was established based on clinical, radiographic, endoscopic, and manometric criteria. The severity of five cardinal symptoms was scored on a scale of 0-3, and each patient received a total symptom score of 1-15. Manometry was subsequently performed, and the mean of five complete pull-through measurements was recorded as the resting LES relaxation pressure. RESULTS: A total of 115 patients (67 male and 48 female) with a mean age of 37.7 yr (range 12-90 yr) were included in the study. The mean total symptom score was 9.32 (range 3.00-14.00) and mean LES relaxation pressure before therapy was 56.29 mm Hg (range 8.00-107.80 mm Hg). Linear regression analysis showed a significant association between the total symptom score and LES relaxation pressure (p < 0.002, r = 0.290). Among the main symptoms, active and passive regurgitation showed significant correlation with LES relaxation pressure when compared to other individual symptoms using Pearson's correlation coefficient (p < 0.001 and 0.002, respectively). CONCLUSIONS: Our study showed that a clinical symptom score can be an appropriate predictor of the LES relaxation pressure in patients with idiopathic achalasia before therapy. Further studies are needed to evaluate similar correlations after therapeutic intervention.     

Psychological Aspects in Children and Adolescents With Major Thalassemia: A Case-Control Study

Background:

Thalassemia is an inherited blood disease. It is a serious public health problem throughout the Mediterranean region, the Middle East and the Indian subcontinent, as well as in Southeast Asia.

Objectives:

Thalassemia is an inherited blood disease. It is a serious public health problem. In this study we assessed psychological aspects in Iranian children and adolescents with thalassemia major.

Patients and Methods:

In this case-control study sixty healthy subjects aged 7-18 years and Sixty Patients with confirmed diagnosis of major thalassemia were enrolled. After obtaining informed consent from parents of all participating thalassemia patients and healthycontrols, we assessed psychological aspects and quality of life by Pediatric Quality of LifeTM (PedsQL™), Strengths and Difficulties Questionnaires (SDQ), State and Trait Anxiety, Children''s Depression Inventory (CDI).

Results:

The results of this study indicate that there are significant changes in depression, anxiety, QOL and behavioral screening between children with thalassemia major compared with healthy subjects by means of both parents and children reports. According to the results, children with thalassemia major have more psychological problems than healthy ones. Patients with thalassemia have a lower QOL than their peers (P = 0.001), the rate of depression is higher in this group (P = 0.015), Also behavioral problems in these children are more than healthy subjects (P = 0.009).

Conclusions:

We recommend appropriate treatment and counseling procedures in addition to specific treatment of thalassemia. According to the results we suggest to establish pediatric psychiatric clinics beside thalassemic clinics to cure psychological aspects of the disease.     

Development of an In-House TaqMan Real Time RT-PCR Assay to Quantify Hepatitis C Virus RNA in Serum and Peripheral Blood Mononuclear Cells in Patients With Chronic Hepatitis C Virus Infection

Background:

Viral load measurements are commonly used to monitor HCV infection in patients with chronic diseases or determining the number of HCV-genomes in serum samples of patients after sustained virological response. However, in some patients, HCV viral load in serum samples is too low to be detected by PCR, especially after treatment.

Objectives:

The aim of this study was to develop a highly specific, sensitive, and reproducible in-house quantitative PCR using specific primers and probe cited in highly conservative region of HCV genome that allows simultaneous detection of HCV genotypes 1 - 4.

Materials and Methods:

In this study, three sets of primer pairs and a TaqMan probe for amplification and detection of selected region within 5’-non-coding (5’NCR) of four HCV genotypes were used. Using plasmid containing 5’NCR region of HCV, standard curve, threshold, and threshold cycle (CT) values were determined. Real-time and nested PCR were performed on HCV genotypes 1 - 4 extracted from plasma and peripheral blood mononuclear cells (PBMCs) samples collected from patients with chronic HCV infection.

Results:

The lower limit detection of this in-house HCV real-time RT-PCR was determined as 100 RNA copies/mL. Inter- and intra-assay coefficient of variation (CV) of this in-house HCV real-time RT-PCR ranged from 0.9% to 1.8% and 1.76% to 3.94%, respectively. The viral load of the genotyped samples ranged from 2.0 × 10⁶ ± 0.31 to 2.7 × 10⁵ ± 0.46 copies/mL in serum samples and 5 × 10² ± 0.36 to 4.0 × 10³ ± 0.51 copies/10⁶ cells/mL of PBMCs.

Conclusions:

The quite sensitive in-house TaqMan real time RT-PCR assay was able to detect and quantify all four main HCV genotypes prevailing around all geographical regions of Iran.     

Serum level of copper in patients with coronary artery disease

Background:

Atherosclerotic disease is the most important cause of mortality in the world. Oxidation is an important pathway in the pathogenesis of coronary artery disease (CAD) through oxidation of low-density lipoprotein (LDL) and free radical formation. Copper (Cu) is an essential micronutrient for enzymes that catalyse LDL oxidation reactions. Therefore, an evaluation of Cu in the atherosclerotic disease is important.

Materials and Methods:

In this study, 334 subjects without recent cardiac event and history of collagen vascular or infectious disease were investigated. All patients divided into four groups to evaluate severity of CAD according to Syntax scoring system. All groups were matched in cardiovascular risk factors.

Results:

The serum level of Cu was significantly higher in total atherosclerotic groups than normal group (P value = 0.001) and significantly increased with severity of atherosclerosis.

Conclusion:

The finding indicated that the serum level of Cu is higher in atherosclerotic patients and it increases with severity of atherosclerosis. Therefore, it may be possible that the basic relationship exist between serum Cu level and atherosclerosis and an association between Cu level and severity of atherosclerosis.     

Insights from a combination of theoretical and experimental methods for probing the biomolecular interactions between human serum albumin and clomiphene

In this study, the interaction of clomiphene (CLO), a non-steroidal and ovulatory stimulant drug employed in the treatment of infertility, with human serum albumin (HSA), the most abundant plasma transport protein, was investigated using spectrofluorometric, FT-IR, UV-Vis, and molecular modeling methods. The obtained results indicated that the binding of CLO to HSA led to intense fluorescence quenching of HSA via a static quenching mechanism, and that the process of CLO binding to HSA was enthalpy driven. By using experimental and theoretical methods, it was confirmed that as a result of binding CLO, slight conformational changes in HSA occurred. Also, the negative ΔH of interaction indicated that the binding of CLO with HSA was mainly enthalpy driven. The experimental and computational results suggested that hydrogen bonds and van der Waals interactions played a major role in the binding, with overall binding constants of K = 3.67 × 10⁹ M⁻¹ at 286 K and 6.52 × 10⁵ mol L⁻¹ at 310 K. Moreover, the results of molecular modeling showed that Asp234, Phe228, Leu327, and Arg209 in HSA had the highest interaction energies with the ligand.

In this study, the interaction of clomiphene with human serum albumin (HSA), the most abundant plasma transport protein, was investigated using spectrofluorometric, FT-IR, UV-Vis, and molecular modeling methods.     

Synthetic ionophores as non-resistant antibiotic adjuvants

Antimicrobial resistance is a world-wide health care crisis. New antimicrobials must both exhibit potency and thwart the ability of bacteria to develop resistance to them. We report the use of synthetic ionophores as a new approach to developing non-resistant antimicrobials and adjuvants. Most studies involving amphiphilic antimicrobials have focused on either developing synthetic amphiphiles that show ion transport, or developing non-cytotoxic analogs of such peptidic amphiphiles as colistin. We have rationally designed, prepared, and evaluated crown ether-based synthetic ionophores (‘hydraphiles’) that show selective ion transport through bilayer membranes and are toxic to bacteria. We report here that hydraphiles exhibit a broad range of antimicrobial properties and that they function as adjuvants in concert with FDA-approved antibiotics against multi-drug resistant (MDR) bacteria. Studies described herein demonstrate that benzyl C₁₄ hydraphile (BC₁₄H) shows high efficacy as an antimicrobial. BC₁₄H, at sub-MIC concentrations, forms aggregates of ∼200 nm that interact with the surface of bacteria. Surface-active BC₁₄H then localizes in the bacterial membranes, which increases their permeability. As a result, antibiotic influx into the bacterial cytosol increases in the presence of BC_nHs. Efflux pump inhibition and accumulation of substrate was also observed, likely due to disruption of the cation gradient. As a result, BC₁₄H recovers the activity of norfloxacin by 128-fold against resistant Staphylococcus aureus. BC₁₄H shows extremely low resistance development and is less cytotoxic than colistin. Overall, synthetic ionophores represent a new scaffold for developing efficient and non-resistant antimicrobial-adjuvants.

Antimicrobial resistance is a world-wide health care crisis.     

The mosquito protein AEG12 displays both cytolytic and antiviral properties via a common lipid transfer mechanism

The mosquito protein AEG12 is up-regulated in response to blood meals and flavivirus infection though its function remained elusive. Here, we determine the three-dimensional structure of AEG12 and describe the binding specificity of acyl-chain ligands within its large central hydrophobic cavity. We show that AEG12 displays hemolytic and cytolytic activity by selectively delivering unsaturated fatty acid cargoes into phosphatidylcholine-rich lipid bilayers. This property of AEG12 also enables it to inhibit replication of enveloped viruses such as Dengue and Zika viruses at low micromolar concentrations. Weaker inhibition was observed against more distantly related coronaviruses and lentivirus, while no inhibition was observed against the nonenveloped virus adeno-associated virus. Together, our results uncover the mechanistic understanding of AEG12 function and provide the necessary implications for its use as a broad-spectrum therapeutic against cellular and viral targets.

Mosquito-borne flaviviruses such as Dengue and Zika represent a major public health risk; 96 million new cases of Dengue infection are reported every year (1), while the Zika infection can cause neurological sequleae and microencephaly (2). Vaccines have had limited success in preventing human flavivirus infections, demonstrating the need for new therapeutic approaches (2). Like humans, mosquitos mount a robust anti-viral immune response against flaviviruses. For example, the mosquito protein AEG12 and its homologs have been shown to be up-regulated in Aedes aegypti in response to Zika virus, West Nile virus, dengue, and Yellow Fever virus infection via JAK-STAT pathways (3–7). AEG12 homologs are also up-regulated upon infection by the parasite Brugia malayi (8), suggesting a role against a wide range of viral- and cell-based targets. Previous sequence analysis of AEG12 revealed it to be a member of the major allergy (MA) protein family, so named because the prototypical protein for this family (Bla g 1) is a major human allergen (9). While many insects contain MA proteins, the family is significantly expanded in Aedes (∼20) and Culex (∼8) mosquitoes; both are major vectors of arboviruses (9, 10). Elucidating the molecular basis for this antiviral activity may provide valuable insight into the development of novel flavivirus and broad-spectrum therapeutic strategies.Curiously, AEG12 was initially described not in the context of flavivirus infection but digestion. Here, AEG12 proteins were found to be highly up-regulated in the midgut of female A. aegypti and subsequently localized to the microvilli following a blood feed but not a sugar feed (9, 11, 12). A similar expression pattern was observed in the AcG12 homolog from Anopheles cilicifacies, suggesting a conserved function across mosquito species (13). In cockroaches, suppression of Bla g 1 expression significantly hindered digestion, resulting in starvation despite adequate food intake suggesting a broader role for MA domain proteins in breaking down and absorbing nutrients from both blood and nonblood food materials (14, 15). However, the function and mechanistic roles of AEG12 proteins remains unknown.The structure of Bla g 1 consists of two consecutive units, each containing five helices (α1 to α5) arranged in a pentagon with the sixth helix (α6) stacking on top. The two pentagonal structures come together to enclose an exceptionally large hydrophobic cavity of ∼3,800 ų that can accommodate up to eight fatty acid ligands (16, 17). Bla g 1 isolated from its natural allergen source was found to contain a mixture of palmitate, oleate, and stearate fatty acids (nMix), the presence of which significantly enhanced Bla g 1 thermostability (16, 17). We hypothesize that these lipid cargoes play a key role in the biological function of both Bla g 1 and AEG12 in digestion and antiviral activity. Indeed, previous studies showed that free fatty acids have viricidal effects against enveloped viruses because of their ability to permeabilize and disrupt their lipid membranes (18–21). Additionally, both free fatty acids and fatty acid–protein conjugates have been shown to destabilize and lyse mammalian cells membranes (22, 23). The ability of AEG12 to mobilize and deliver a range of fatty acid cargoes could provide a common mechanism through which it fulfills both putative functions.Here, we demonstrate that AEG12 adopts a similar structure as Bla g 1, enabling it to bind a range of fatty acid ligands within its central hydrophobic cavity. Selective delivery of these fatty acid cargoes into phosphatidylcholine (PC) lipid bilayers results in membrane destabilization and cytotoxicity against eukaryotic cells consistent with a role in erythrocyte digestion. Similarly, AEG12 displayed antiviral activity against a range of enveloped, but not nonenveloped viruses suggesting that both putative functions of AEG12 share a common mechanism of action centered on lipid exchange resulting in membrane destabilization.     

Evaluation of corneal thickness alterations during menstrual cycle in productive age women

Purpose:

To determine the change in corneal thickness through different phases of menstrual cycle in women who are in their productive age.

Materials and Methods:

Fifty healthy women with normal past medical history were enrolled in this prospective study. Central corneal thickness was measured with ultrasound pachymeter three times during a menstrual cycle: Beginning of the cycle (days 1-3), ovulation time, and at the end of cycle (days 27-32). We confirmed ovulation time with determining a peak in luteinizing hormone in urine. To avoid the diurnal variation of the corneal thickness which is well recognized, we checked all our subjects at 10 in the morning.

Results:

In days 1 to 3 of menstruation, mean corneal thickness was 541.40±11.36 and 540.82±11.70 microns for left and right eyes respectively. At ovulation time the mean thickness changed to 556.50±7.11 and 555.98±7.26 microns for left and right eyes respectively, and at the end of the cycle, the corneal thickness turned in to 536.38±12.83 and 535.48±13.08 microns for left and right eyes respectively. The difference of corneal thickness was statistically significant relating to the different stages of menstrual cycle.

Conclusion:

The thickest cornea during the menstruation cycle is achieved at the ovulation time and the thinnest at the end of the cycle and this should be taken in to account whilst plan to do a corneal refractive surgery.