Knowledge and Practice of Healthcare Professionals in the Medical Care of Asthma Adult Patients in Jordan with a Particular Reference to Adherence to GINA Recommendations.

Purpose: This study aimed to evaluate Jordanian healthcare professionals' knowledge, skills, and experience in dealing with and treating adult asthmatic patients and assess healthcare professionals' compliance with the Global Initiative for Asthma (GINA) guideline recommendations for asthma treatment and management in Jordan. Patients and Methods: A cross-sectional survey was conducted in Jordan between May and June 2023. This study included physicians, pharmacists, and nurses working in clinical settings and directly interacting with asthma patients in Jordan's private and governmental healthcare facilities who were eligible to participate in this research. Three primary sections of an online survey were constructed using Google Forms and included demographic data, a diagnosis and management of the illness, and a list of worldwide medical references for asthma. Results: A total of 300 healthcare professionals, including pharmacists (103), nurses (87), Doctor of Pharmacy (28), and physicians (82), took part in this research. The survey respondents' general knowledge of the nature and symptoms of asthma is quite high, exceeding 84% and 86%, respectively. The survey also showed a high degree of awareness of asthma diagnosis. It was also revealed that around one-quarter of participants consistently reported the severity of asthma. The most frequent factor influencing the decision to take controller or maintenance medicines is symptom severity (reported from 86% of participants). The GINA guidelines were selected as the asthma information source by almost half of the participants. Finally, 47% of participants were aware of the GINA's 2019 adjustments of the recommendations. Conclusion: This study showed that Jordanian healthcare professionals understand the nature, symptoms, and diagnoses of asthma disease. However, many of the survey participants lack adherence, awareness, and understanding of the GINA recommendations about asthma management. More research is required to assess the adherence to the evidence-based recommendations and evaluate the impact of training courses.

The role of human rhinovirus in COPD exacerbations in Abu Dhabi: molecular epidemiology and clinical significance.

This study aimed to describe the molecular epidemiology and seasonality of human rhinovirus (HRV) in chronic obstructive pulmonary disease (COPD) and its association with COPD exacerbations in Abu Dhabi, the United Arab Emirates (UAE). Sputum specimens were collected for analysis from all COPD patients who visited a medical center from November 2021 to October 2022. The real-time quantitative polymerase chain reaction (qPCR) test was used to detect HRV. Of the 78 COPD patients included in the study, 58 (74%) patients presented with one or more exacerbation episodes. The incidence of COPD exacerbation peaked over the winter and substantially decreased during the summer. HRV positivity in patients during exacerbation (E1) was 11/58 (19%) and 15/58 (26%) two weeks after the exacerbation episode (E2). There was no significant difference in the HRV load in these patients. No statistically significant difference was observed in the detection of HRV during exacerbation compared to patients with stable COPD. This is the first study to assess the association between HRV detection by qPCR and COPD exacerbations in the UAE. The high sensitivity of the detection technology helped collect reliable epidemiologic data. Few studies have provided similar Middle East data. This study's pattern of COPD exacerbations and HRV detection parallels that of temperate countries. This information can help with future, more extensive surveillance of respiratory viruses in the UAE and the Middle East and their association with COPD exacerbations.

Evaluation of the expression levels of lncRNAs H19 and MEG3 in patients with type 2 diabetes mellitus.

BACKGROUND: LncRNAs may play a role in either suppressing or exacerbating diabetes-associated vascular complications. AIMS: This study aimed to assess MEG3 and H19 expression levels in T2DM and pre-diabetes and their roles in diabetes-related microvascular complications. SUBJECT AND METHODS: (RT-PCR) analysis of the MEG3 and H19 plasma levels was carried out in 180 participants of T2DM, pre-diabetes, and control. RESULTS: The expression level of lncRNA H19 was significantly down-regulated and lncRNA MEG3 up-regulated in T2DM compared to pre-diabetes and control, also for pre-diabetes versus control. The (ROC) analysis of MEG3 and H19 relative expression levels showed that MEG3 has better sensitivity for distinguishing T2DM from pre-diabetes and control groups.In comparison, H19 offered superior sensitivity to distinguish pre-diabetic from controls. Additionally, H19 was reported as an independent risk factor for T2DM by multivariate analysis. Low expression of H19 and over-expressed MEG3 were significantly associated with retinopathy, nephropathy, and elevated renal indicators (urea, creatinine, and UACR. CONCLUSION: Our results implicated the potential diagnostic and predictive roles of lncRNA MEG3 and H19 for T2DM and related microvascular complications. Additionally, H19 may serve as a potential biomarker for pre-diabetes prediction.

Co-infection of COVID-19 patients with atypical bacteria: A study based in Jordan.

Objective: The aim of this work was to know the prevalence of Chlamydophila pneumoniae and Mycoplasma pneumoniae in coronavirus disease 2019 (COVID-19) patients in Jordan. Also, to assess a TaqMan real-time polymerase chain reaction (PCR) assay in detecting these two bacteria. Methods: This is a retrospective study performed over the last five months of the 2021. All nasopharyngeal specimens from COVID-19 patients were tested for C. pneumonia, and M. pneumoniae. The C. pneumoniae Pst-1 gene and M. pneumoniae P1 cytadhesin protein gene were the targets. Results: In this study, 14 out of 175 individuals with confirmed severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection (8.0%) were co-infected with C. pneumoniae or M. pneumoniae. Co-infection with SARS-CoV-2 and C. pneumoniae was reported in 5 (2.9%) patients, while 9 (5.1%) patients had M. pneumoniae and SARS-CoV-2 co-infection. The mean (± std) of the correlation coefficient of the calibration curve for real-time PCR analysis was -0.993 (± 0.001) for C. pneumoniae and -0.994 (± 0.003) for M. pneumoniae. The mean amplification efficiencies of C. pneumoniae and M. Pneumoniae were 187.62% and 136.86%, respectively. Conclusion: In this first study based in Jordan, patients infected with COVID-19 have a low rate of atypical bacterial co-infection. However, clinicians should suspect co-infections with both common and uncommon bacteria in COVID-19 patients. Large prospective investigations are needed to give additional insight on the true prevalence of these co-infections and their impact on the clinical course of COVID-19 patients.

Co-infection of COVID-19 patients with atypical bacteria: A study based in Jordan

Objective: The aim of this work was to know the prevalence of Chlamydophila pneumoniae and Mycoplasma pneumoniae in coronavirus disease 2019 (COVID-19) patients in Jordan. Also, to assess a TaqMan real-time polymerase chain reaction (PCR) assay in detecting these two bacteria. Methods: This is a retrospective study performed over the last five months of the 2021. All nasopharyngeal specimens from COVID-19 patients were tested for C. pneumonia, and M. pneumoniae. The C. pneumoniae Pst-1 gene and M. pneumoniae P1 cytadhesin protein gene were the targets. Results: In this study, 14 out of 175 individuals with confirmed severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection (8.0%) were co‐infected with C. pneumoniae or M. pneumoniae. Co‐infection with SARS‐CoV‐2 and C. pneumoniae was reported in 5 (2.9%) patients, while 9 (5.1%) patients had M. pneumoniae and SARS‐CoV‐2 co-infection. The mean (± std) of the correlation coefficient of the calibration curve for real-time PCR analysis was –0.993 (± 0.001) for C. pneumoniae and –0.994 (± 0.003) for M. pneumoniae. The mean amplification efficiencies of C. pneumoniae and M. Pneumoniae were 187.62% and 136.86%, respectively. Conclusion: In this first study based in Jordan, patients infected with COVID-19 have a low rate of atypical bacterial co-infection. However, clinicians should suspect co-infections with both common and uncommon bacteria in COVID-19 patients. Large prospective investigations are needed to give additional insight on the true prevalence of these co-infections and their impact on the clinical course of COVID-19 patients. (AU)

Molecular Accounting and Profiling of Human Respiratory Microbial Communities: Toward Precision Medicine by Targeting the Respiratory Microbiome for Disease Diagnosis and Treatment.

The wide diversity of microbiota at the genera and species levels across sites and individuals is related to various causes and the observed differences between individuals. Efforts are underway to further understand and characterize the human-associated microbiota and its microbiome. Using 16S rDNA as a genetic marker for bacterial identification improved the detection and profiling of qualitative and quantitative changes within a bacterial population. In this light, this review provides a comprehensive overview of the basic concepts and clinical applications of the respiratory microbiome, alongside an in-depth explanation of the molecular targets and the potential relationship between the respiratory microbiome and respiratory disease pathogenesis. The paucity of robust evidence supporting the correlation between the respiratory microbiome and disease pathogenesis is currently the main challenge for not considering the microbiome as a novel druggable target for therapeutic intervention. Therefore, further studies are needed, especially prospective studies, to identify other drivers of microbiome diversity and to better understand the changes in the lung microbiome along with the potential association with disease and medications. Thus, finding a therapeutic target and unfolding its clinical significance would be crucial.

Combined Effect of Omega-3 Fatty Acid and Vitamin D 3 on Oxidized LDL-C and Non-HDL-C Levels in People With Vitamin D Deficiency: A Randomized Controlled Trial.

ABSTRACT: The present randomized clinical trial (RCT) was conducted on Jordanian participants with vitamin D deficiency (VDD) with no other medical conditions, to evaluate the combined effect of 1,25-dihydroxy vitamin D 3 (Vit.D 3 ) and omega-3 fatty acid (n-3FA) supplements (D+) on oxidized low-density lipoprotein (Ox-LDL) and non-high-density lipoprotein cholesterol (non-HDL-C) levels as common predictors of cardiovascular diseases (CVDs). Participants were randomized into 4 groups as follows: a control group (C) that received no supplementations, a Vit.D 3 group that received 50,000 IU of Vit.D 3 every week, an n-3FA group that received 300 mg of omega-3 fatty acid every day, and a D+ group that received a combination of both supplements, with the same dosage administered by the previous groups but with a 4-6-hour time interval between Vit.D 3 and n-3FA administration to avoid any possible interaction. All supplementations were administered orally for 8 weeks. Forty-seven participants were allocated to each group. Twenty-six in the control group, 37 participants in the Vit.D 3 group, 37 participants in the n-3FA group, and 46 participants in the D+ group completed the study to the end. The D+ supplementations significantly increased non-HDL-C (118.99 ± 60.98 to 155.26 ± 43.36 mg/dL, P << 0.05) but decreased Ox-LDL-C levels (69.29 ± 37.69 to 52.81 ± 17.30 pg/mL, P = 0.03). The stepwise regression showed that the serum LDL-C level was the main independent variable involved in the elevation of non-HDL levels (R 2 = 0.837) observed at the end of the trial in the D+ group. The groups that were supplemented with either Vit.D 3 alone or n-3FA alone had an insignificant decrease in the level of Ox-LDL-C. In conclusion, despite the observed hyperlipidemic effect, the combination treatment is recommended by the research team because the decrease in Ox-LDL may offset the hyperlipidemic effect.

Ethanol-induced dose dumping from sodium alginate matrix tablets: Investigation of the effects of medium viscosity and pH.

In this work, the swelling and disintegration of drug-free sodium alginate (SA) compacts and the release of metformin HCl from SA matrix tablets were investigated in acidic media of different ethanol concentrations (0, 10, 20, and 40 % v/v), pH (1.2 and 4.5) and HPMC K4M concentrations (0-1 % w/v). The investigated dissolution media represented the consumption of different alcoholic beverages, the pH of fasted and fed states, and a range of viscosity resembling diluted homogenized FDA meal. The dissolution efficiency and the time to 50 % release (t50%) were selected as release parameters. It was found that both ethanol concentration and medium pH affected drug release from SA matrix tablets and the swelling of SA compacts. Dose dumping occurred at high ethanol concentration (40 %) at both media pH with almost complete drug release within 15-30 min associated with rapid matrix disintegration. HPMC at 0.5-1 % concentrations increased the medium's viscosity, preventing dose dumping at high ethanol concentrations. Erosion and disintegration of SA compacts were decelerated by increasing HPMC concentration in hydroethanolic media in consonance with decreased release rate from matrix tablets. ANOVA tests showed significant effects of pH and concentrations of ethanol and HPMC in the dissolution medium on the release parameters.

Bovine Colostrum Treatment of Specific Cancer Types: Current Evidence and Future Opportunities.

Worldwide, the incidence of cancer is on the rise. Current cancer treatments include chemotherapy, radiation therapy, and surgery. Chemotherapy and radiation treatment are typically associated with severe adverse effects and a decline in patients' quality of life. Anti-cancer substances derived from plants and animals need to be evaluated therapeutically as it is cost-effective, have fewer side effects, and can improve cancer patients' quality of life. Recently, bovine colostrum (BC) has attracted the interest of numerous researchers investigating its anti-cancer potential in humans. Dressings loaded with BC are beneficial in treating chronic wounds and diabetic foot ulcers. Lactoferrin, a glycoprotein with potent anti-oxidant, anti-inflammatory, anti-cancer, and anti-microbial effects, is abundant in BC. The BC pills successfully promote the regression of low-grade cervical intraepithelial neoplasia when administered intravaginally. The biological, genetic, and molecular mechanisms driving BC remain to be determined. Oral BC supplements are generally well-tolerated, but some flatulence and nausea may happen. To evaluate the therapeutic effects, long-term safety, and appropriate dosages of BC drugs, well-designed clinical trials are necessary. The purpose of this article is to emphasize the anti-cancer potential of BC and its constituents.

Evaluation of the effectiveness of educational medical informatics tutorial on improving pharmacy students' knowledge and skills about the clinical problem-solving process.

Objective: To investigate the effectiveness of an online tutorial and its impact on improving knowledge and skills of pharmacy students in the clinical problem-solving process that is necessary to implement pharmaceutical care. Methods: This is a prospective interventional study conducted during the COVID-19 pandemic restrictions using four novel templates. The first two levels of Kirkpatrick's Model (Reaction and Learning) were used. Results: 129 participants completed all of the online training parts. The findings indicated a significant improvement in the students' knowledge and skills. The participants achieved higher score following the tutorial than the baseline, with a statistically significant difference (p < 0.001). There was a significant improvement in the number of detected treatment-related problems. The majority of students were satisfied with the overall training process and stated a high evaluation score out of 10 (mean = 7.93 ± 1.42, median = 8.00). Conclusion: The educational intervention achieved a substantial positive impact on decision-making skills of participating students and was considered effective in helping them attain basic skills such as teamwork, peer assessment, communication and critical evaluation. Healthcare providers must work together to ensure accurate medication use during care transitions. Pharmacists, as medication experts, play an important role in the implementation process. Pharmacy educators must prepare pharmacy student to use pharmaceutical care in their future practice.

Comparison between clinical characteristics and laboratory findings among patients with complicated and noncomplicated SARS-CoV-2 infection: A single-center experience from Shebin Al-Kom, Egypt.

BACKGROUND: Coronavirus disease 2019 (COVID-19) infection is considered a serious highly infectious disease caused by severe acute respiratory syndrome coronavirus 2, resulting in more than 6.27 million deaths worldwide. AIM OF THE STUDY: The study aimed to compare clinical characteristics and laboratory findings of COVID-19 patients with complications and without complications and discriminate the important risk factors for the complications and deaths. SUBJECTS AND METHODS: This cross-sectional study included 75 confirmed COVID-19 positive patients; out of which 49 were severely-ill cases. Analysis of all patients' clinical and laboratory information on admission including serum ferritin, thrombotic activity (d-dimer), lactate dehydrogenase (LDH), C-reactive protein (CRP), creatinine, aspartate aminotransferase, and alanine aminotransferase were done. RESULTS: Lymphopenia, tachycardia, tachypnea, elevated CRP, d-dimer, serum ferritin, LDH, and decreased SpO2 were significantly associated with complicated cases (p < .05 for all). By using multivariate logistic regression analysis models, elevated serum ferritin and tachycardia were significantly correlated with the increased odds of complicated COVID-19 cases (odds ratio [confidence interval 95%] = 10.42 [2.32-46.89] and 8.01 [1.17-55.99]; respectively) (p = .002 and .007, respectively). CONCLUSION: Lymphocytopenia, d-dimer, LDH, and CRP levels, which were significantly linked to the severity of COVID-19, were the prognostic biomarkers to predict the disease severity.

Perspectives of the community in the developing countries toward telemedicine and pharmaceutical care during the COVID-19 pandemic.

Background: Pharmaceutical care (PC) services have expanded in recent years, resulting in improved patient outcomes. However, such PC services are currently available for free in the majority of Arabic countries. During the coronavirus disease (COVID-19) pandemic, telemedicine is especially beneficial since it allows for continuity of care while allowing for social distancing and minimizing the risk of infection. Objective: To assess the community's attitude, opinion, and willingness to pay for telemedicine and PC services during COVID-19 pandemic, as well as to create a website provision for telemedicine and PC services. Methods: This cross-sectional study was conducted, over five months (December 2020- April 2021), among the general population in Arabic countries, excluding pharmacists, physicians, and pharmacy students. Results: A total of 1717 participants were involved, most of them were from Jordan (52.2%) and Iraq (24.8%). Sixty two percent of participants seek pharmacists' advice whenever they have any medication changes and 45.1% of the participants agreed with the idea of paying pharmacists to decrease medication errors. Interestingly, 89.5% of participants encouraged the idea of creating a website that provides a PC, and 35.5% of them would pay for it. The failure to document the medical information of the patients had most applicants' agreement as a reason of medical errors (M=4.17/5, SD=0.787). More than three-quarters of participants agreed that creating a database containing the patients' medical information will reduce medical errors. Conclusion: From a patients' perspective, this study suggests a large patient need for expanding PC services in Arabic countries and introduces a direct estimate of the monetary value for the PC services to contribute to higher savings. The majority of participants supported the idea of creating a website provision of telemedicine and PC services, and a considerable proportion of them agreed to pay for it.

Evaluation of the effectiveness of educational medical informatics tutorial on improving pharmacy students’ knowledge and skills about the clinical problem-solving process

Objective: To investigate the effectiveness of an online tutorial and its impact on improving knowledge and skills of pharmacy students in the clinical problem-solving process that is necessary to implement pharmaceutical care. Methods: This is a prospective interventional study conducted during the COVID-19 pandemic restrictions using four novel templates. The first two levels of Kirkpatrick’s Model (Reaction and Learning) were used. Results: 129 participants completed all of the online training parts. The findings indicated a significant improvement in the students’ knowledge and skills. The participants achieved higher score following the tutorial than the baseline, with a statistically significant difference (p < 0.001). There was a significant improvement in the number of detected treatment-related problems. The majority of students were satisfied with the overall training process and stated a high evaluation score out of 10 (mean = 7.93 ± 1.42, median = 8.00). Conclusion: The educational intervention achieved a substantial positive impact on decision-making skills of participating students and was considered effective in helping them attain basic skills such as teamwork, peer assessment, communication and critical evaluation. Healthcare providers must work together to ensure accurate medication use during care transitions. Pharmacists, as medication experts, play an important role in the implementation process. Pharmacy educators must prepare pharmacy student to use pharmaceutical care in their future practice. (AU)

Perspectives of the community in the developing countries toward telemedicine and pharmaceutical care during the COVID-19 pandemic

Background: Pharmaceutical care (PC) services have expanded in recent years, resulting in improved patient outcomes. However, such PC services are currently available for free in the majority of Arabic countries. During the coronavirus disease (COVID-19) pandemic, telemedicine is especially beneficial since it allows for continuity of care while allowing for social distancing and minimizing the risk of infection. Objective: To assess the community’s attitude, opinion, and willingness to pay for telemedicine and PC services during COVID-19 pandemic, as well as to create a website provision for telemedicine and PC services. Methods: This cross-sectional study was conducted, over five months (December 2020– April 2021), among the general population in Arabic countries, excluding pharmacists, physicians, and pharmacy students. Results: A total of 1717 participants were involved, most of them were from Jordan (52.2%) and Iraq (24.8%). Sixty two percent of participants seek pharmacists’ advice whenever they have any medication changes and 45.1% of the participants agreed with the idea of paying pharmacists to decrease medication errors. Interestingly, 89.5% of participants encouraged the idea of creating a website that provides a PC, and 35.5% of them would pay for it. The failure to document the medical information of the patients had most applicants’ agreement as a reason of medical errors (M=4.17/5, SD=0.787). More than three-quarters of participants agreed that creating a database containing the patients’ medical information will reduce medical errors. Conclusion: From a patients’ perspective, this study suggests a large patient need for expanding PC services in Arabic countries and introduces a direct estimate of the monetary value for the PC services to contribute to higher savings. The majority of participants supported the idea of creating a website provision of telemedicine and PC services, and a considerable proportion of them agreed to pay for it. (AU)

Umbelliferone alleviates hepatic ischemia/reperfusion-induced oxidative stress injury via targeting Keap-1/Nrf-2/ARE and TLR4/NF-κB-p65 signaling pathway.

Umbelliferone (UMB; 7-hydroxycoumarin) is a natural compound that exhibited a diversity of pharmacological activities. Its protective effects against various ischemia/reperfusion (IR) injuries, including heart, kidney, and testis, have been observed. However, their effect on hepatic IR is still not investigated yet. Here, this study was conducted to examine the potential protective role of UMB during the early phase of hepatic IR injury via targeting Keap-1/Nrf-2/ARE and its closely related signaling pathway, TLR4/NF-κB-p65. Experimentally, forty Wistar albino rats were randomly divided into 4 groups: Sham control group (received 1% carboxymethyl cellulose as a vehicle), UMB group (30 mg/kg/day, P.O.), IR group (subjected to complete hepatic IR injury), and IR + UMB group. Our results revealed that oral UMB effectively reduced the serum levels of ALT, AST, ALP, and LDH along with the restoration of oxidant/antioxidant status. At the molecular level, UMB markedly activated Nrf-2 expression and its down-streaming targets: HO-1, NQO1, GCLC, SOD3, and TNXRD1, along with Keap-1 down-regulation. Besides, UMB significantly down-regulated NF-κB-p65 and TLR4 expressions with subsequent decreased TNF-α and IL-1ß levels coupled with the up-regulation of the IL-10 level. Finally, biochemical findings were confirmed by attenuation of histopathological changes in liver tissues. Together, UMB is a promising agent for the amelioration of liver tissues against IR-induced oxidative injury through activation of the Keap-1/Nrf-2/ARE signaling pathway along with suppression of its closely related signaling pathways: TLR4/NF-κB-p65. Illustrated diagram explored the prospective underlying protective mechanism of UMB against IR-induced hepatic damage.

A Cross-Sectional Study on the Combined Effect of Body Weight and Coffee Consumption on Serum Levels of Leptin, Vitamin B12, and Folic Acid in Healthy Young Adult Males.

PURPOSE: Studies on the effect of body weight and coffee consumption on leptin, vitamin B12, and folic acid are scarce and conflicting. This study investigates the effect of body weight and/or coffee consumption rate on the serum levels of these molecules in healthy young adult males. PATIENTS AND METHODS: This observational cross-sectional study was carried out at the faculty of pharmacy, Applied Science Private University (ASU), Amman, Jordan, from July to September 2020. Young healthy males were invited to participate in the study and fill a questionnaire regarding lifestyle habits including coffee consumption during the last 3 months, medical history, and anthropometric measurements. Depending on BMI and extent of coffee consumption, participants were divided into 4 groups; normal body weight and moderate coffee consumption (NW/MCC) group; normal body weight and heavy coffee consumption (NW/HCC) group; overweight and moderate coffee consumption (OW/MCC) group; overweight and heavy coffee consumption (OW/HCC) group. Serum samples were taken to measure leptin, vitamin B12, and folic acid levels in addition to morning and midnight salivary cortisol and dehydroepiandrosterone (DHEA) samples. RESULTS: Healthy males (n = 122) aged 18 to 26 years continued participation in this study. Serum levels of leptin in NW/MCC, NW/HCC, OW/MCC, OW/HCC groups were 5.93, 5.75, 14.86, 16.79 ng/mL, respectively. Serum levels of vitamin B12 in these groups were 356.09, 402.71, 334.25, 331.05 pg/mL, respectively. While, the serum levels of folic acid were 8.92, 10.27, 10.12, 10.47 ng/mL, respectively. Body weight was positively associated with leptin (p = 0.00), negatively associated with vitamin B12 (p = 0.047), and not associated with folic acid (p = 0.235). Coffee consumption rate had no significant effect on leptin, vitamin B12, or folic acid. Finally, the combination of body weight and coffee consumption had no significant effect on leptin, vitamin B12, or folic acid. CONCLUSION: There was no possible synergistic effect between body weight and coffee consumption rate on leptin, vitamin B12, or folic acid levels. However, overweight was associated with higher leptin, lower vitamin B12, and no change in folic acid levels. TRIAL REGISTRATION: This trial was registered at clinicaltrials.gov as NCT04488731.

ABCB1 polymorphisms and steroid treatment in children with idiopathic nephrotic syndrome.

BACKGROUND: The most common cause of nephrotic syndrome (NS) is idiopathic nephrotic syndrome (INS), also called nephrosis. Although most patients respond to steroid therapy, there is unequal response to treatment suggesting the involvement of genetic factors. The current study was conducted to evaluate the influence of two single nucleotide polymorphisms (SNPs) in ABCB1 (C3435T and C1236T) on the steroid treatment response in INS children. MATERIALS AND METHODS: Genotyping of ABCB1 C3435T and C1236T polymorphisms by real time PCR were conducted on 120 INS children, 80 steroid sensitive (SS) and 40 steroid resistant (SR). RESULTS: A significant difference in the distribution of ABCB1 C3435T and C1236T genotypes was observed between SS and SR patients. C1236T polymorphism was associated with steroid resistance in INS children (odds ratio: 2.27, 95 % confidence interval: 1.2-4.4; P = 0.012). The frequency of the T allele was significantly higher in SR than in SS patients (81.2 vs. 65.6%, respectively). The odds ratio for the C3435T polymorphism in response to steroid treatment was smaller than that of the polymorphism C1236T, and did not reach statistical significance (odds ratio: 1.1, 95 % confidence interval: 0.6-1.9; P = 0.77). CONCLUSION: Our results suggested that C1236T polymorphism in ABCB1 gene was associated with steroid resistance. A higher proportion of SR children had C1236T TT genotype and T allele, these patients may require other therapeutic strategies.

Hypoxia-induced increases in glucose uptake do not cause oxidative injury or advanced glycation end-product (AGE) formation in vascular endothelial cells.

An increase in glucose uptake by endothelial cells exposed to hyperglycemia is the presumed initiating event that causes systemic vascular disease in individuals with diabetes. Diabetics do not develop clinically significant pulmonary vascular disease, however, despite the pulmonary circulation's exposure to the same level of glucose. We hypothesized that pulmonary artery endothelial cells are protected from the detrimental effects of hyperglycemia because they take up less glucose than endothelial cells in the systemic circulation, either because of intrinsic differences between the two cell types or because the lower oxygen tension in the pulmonary arterial blood depresses glucose uptake. To test this hypothesis, we exposed normoglycemic and hyperglycemic bovine pulmonary artery (PAECs) and aortic endothelial cells (AECs) from the same animal to progressively lower oxygen tensions and determined glucose uptake. In contrast with our initial hypothesis, we detected no significant difference in glucose uptake between the two cell types. Furthermore, glucose uptake in both PAECs and AECs increased, not decreased, as the oxygen tension dropped; this oxygen-dependent increase in glucose uptake in endothelial cells predominated over the hyperglycemia-mediated decrease in glucose uptake that has been reported by others. Despite the increase in glucose uptake at lower oxygen tensions, we detected no corresponding increase in protein carbonylation or advanced glycation endproducts. These results demonstrate that small physiologically relevant changes in oxygen tension can have an important impact on glucose uptake in endothelial cells. These results also demonstrate that an increase in glucose uptake, by itself, is not sufficient to generate ROS-mediated protein carbonylation or increase intracellular advanced glycation endproducts in vascular endothelial cells.

Overlap of Doxycycline Fluorescence with that of the Redox-Sensitive Intracellular Reporter roGFP.

Tetracycline-inducible systems allow for either suppression or induction of transgene expression to facilitate studies of cell physiology. Doxycycline is a preferred inducer for these gene expression systems due to its membrane permeability; however, the heterocyclic structure of doxycycline exhibits fluorogenic properties that can potentially bias measurement of other fluorochromes. Thus the simultaneous use of tetracycline-inducible systems and fluorescent proteins as reporter genes or as intracellular biosensors may lead to potentially confounding results. Herein, using cells which co-express the ratiometric redox sensitive intracellular reporter, roGFP, and a tetracycline-inducible reporter plasmid encoding the reporter gene, mCherry, as a model system, we describe the overlapping intracellular fluorescent signals between doxycycline and commonly used intracellular fluorescent probes. In our cells, the addition of doxycycline to cells caused a dose- and time-dependent increase in cell fluorescence with 405 nm excitation which overlapped with that of the oxidized configuration of roGFP. Incubating cells in concentrations of doxycycline less than 1 µg/mL and removing doxycycline from the media 60 min before performing experiments eliminated fluorescence interference while still maintaining maximal reporter transgene activation.

Serum can overcome contact inhibition in confluent human pulmonary artery smooth muscle cells.

Pulmonary artery endothelial cells (PAEC) in an intact vessel are continually exposed to serum, but unless injured, do not proliferate, constrained by confluence. In contrast, pulmonary artery smooth muscle cells (PASMC) attain, and maintain, confluence in the presence of minimal serum, protected from serum's stimulatory effects except when the endothelial barrier becomes more permeable. We hypothesized therefore, that confluent PASMC may be less constrained by contact inhibition in the presence of serum than PAEC and tested this idea by exposing confluent non-transformed human PAEC and PASMC to media containing increasing concentrations of fetal bovine serum (FBS) and determining cell growth over 7 days. PAEC that had attained confluence in low serum did not proliferate even when exposed to 5% serum, the highest concentration tested. In contrast, PASMC that attained confluence in low serum did proliferate once serum levels were increased, an effect that was dose dependent. Consistent with this observation, PASMC had more BrdU incorporation and a greater percentage of cells in S phase in 5% compared to 0.2% FBS, whereas no such difference was seen in PAEC. These results suggest that confluent human PAEC are resistant to the stimulatory effects of serum, whereas confluent PASMC can proliferate when serum levels are increased, an effect mediated in part by differences in phosphoinositide 3-kinase activation. This observation may be relevant to understanding the PASMC hyperplasia observed in humans and animals with pulmonary hypertension in which changes in endothelial permeability due to hypoxia or injury expose the underlying smooth muscle to serum.