The Rapid and Sensitive Quantitative Determination of Galactose by Combined Enzymatic and Colorimetric Method: Application in Neonatal Screening

The quantitative measurement of galactose in blood is essential for the early diagnosis, treatment, and dietary monitoring of galactosemia patients. In this communication, we aimed to develop a rapid, sensitive, and cost-effective combined method for galactose determination in dry blood spots. This procedure was based on the combination of enzymatic reactions of galactose dehydrogenase (GalDH), dihydrolipoyl dehydrogenase (DLD), and alkaline phosphates with a colorimetric system. The incubation time and the concentration of enzymes used in new method were also optimized. The analytical performance was studied by the precision, recovery, linearity, and sensitivity parameters. Statistical analysis was applied to method comparison experiment. The regression equation and correlation coefficient (R2) were Y = 0.0085x + 0.032 and R2 = 0.998, respectively. This assay exhibited a recovery in the range of 91.7–114.3 % and had the limit detection of 0.5 mg/dl for galactose. The between-run coefficient of variation (CV) was between 2.6 and 11.1 . The within-run CV was between 4.9 and 9.2 . Our results indicated that the new and reference methods were in agreement because no significant biases exist between them. Briefly, a quick and reliable combined enzymatic and colorimetric assay was presented for application in newborn mass screening and monitoring of galactosemia patients. © 2016 Springer Science+Business Media New Yor

Cloning and expression of codon-optimized recombinant darbepoetin alfa in Leishmania tarentolae T7-TR

Darbepoetin alfa is an engineered and hyperglycosylated analog of recombinant human erythropoietin (EPO) which is used as a drug in treating anemia in patients with chronic kidney failure and cancer. This study desribes the secretory expression of a codon-optimized recombinant form of darbepoetin alfa in Leishmania tarentolae T7-TR. Synthetic codon-optimized gene was amplified by PCR and cloned into the pLEXSY-I-blecherry3 vector. The resultant expression vector, pLEXSYDarbo, was purified, digested, and electroporated into the L. tarentolae. Expression of recombinant darbepoetin alfa was evaluated by ELISA, reverse-transcription PCR (RT-PCR), Western blotting, and biological activity. After codon optimization, codon adaptation index (CAI) of the gene raised from 0.50 to 0.99 and its GC content changed from 56 to 58. Expression analysis confirmed the presence of a protein band at 40 kDa. Furthermore, reticulocyte experiment results revealed that the activity of expressed darbepoetin alfa was similar to that of its equivalent expressed in Chinese hamster ovary (CHO) cells. These data suggested that the codon optimization and expression in L. tarentolae host provided an efficient approach for high level expression of darbepoetin alfa. © 2015 Elsevier Ltd. All rights reserved

Purification and Characterization of Recombinant Darbepoetin Alfa from Leishmania tarentolae

Darbepoetin alfa is a biopharmaceutical glycoprotein that stimulates erythropoiesis and is used to treat anemia, which associated with renal failure and cancer chemotherapy. We herein describe the structural characterization of recombinant darbepoetin alfa produced by Leishmania tarentolae T7-TR host. The DNA expression cassette was integrated into the L. tarentolae genome through homologous recombination. Transformed clones were selected by antibiotic resistance, diagnostic PCRs, and protein expression analysis. The structure of recombinant darbepoetin alfa was analyzed by isoelectric focusing, ultraviolet–visible spectrum, and circular dichroism (CD) spectroscopy. Expression analysis showed the presence of a protein band at 40 kDa, and its expression level was 51.2 mg/ml of culture medium. Darbepoetin alfa have 5 isoforms with varying degree of sialylation. The UV absorption and CD spectra were analogous to original drug (Aranesp), which confirmed that the produced protein was darbepoetin alfa. Potency test results revealed that the purified protein was biologically active. In brief, the structural and biological characteristics of expressed darbepoetin alfa were very similar to Aranesp which has been normally expressed in CHO. Our data also suggest that produced protein has potential to be developed for clinical use. © 2016, Springer Science+Business Media New York

Silent osteonecrosis of the femoral head following high-dose corticosteroids in patients with systemic rheumatic diseases

Background: Osteonecrosis (ON) is known to be one of the most disabling complications following corticosteroid (CS) medications. However, evidence regarding risk of asymptomatic prevalence of ON among different diseases and the impact of variable steroid regimens are conflicting. We aimed to determine the prevalence of ON of femoral head in asymptomatic patients with systemic rheumatic diseases who received high-dose CS and also clarify its relationship with different dosages and regimens. Methods: In this cross-sectional study, 50 consecutive patients receiving high-dose CS for rheumatic diseases who have no pelvic pain were recruited. MRI of both hips was performed on all patients using a 1.5 Tesla to diagnose ON. Results: Of 50 subjects, 18 (36) developed ON of the femoral head. Groups with and without ON were comparable in terms of sex, age and mean starting CS dose. There was no statistical difference in the type of CS regimen including daily dose, peak dose and cumulative dose between the two groups. However, silent ON was associated with both the cumulative CS dose and the duration of CS therapy. Conclusion: According to high prevalence of ON in our selected patients with no other identifiable risk factor for ON, monitoring of high risk patients with periodic hip MRI would help diagnose necrosis in early stage

The Effect of G-ORS Along With Rice Soup in the Treatment of Acute Diarrhea in Children: A Single-Blind Randomized Controlled Trial

Background: The world health organization guidelines for treatment of diarrhea in children emphasize on continued feeding together with prescription of oral rehydration solution (ORS) and supplementary zinc therapy. However, conflicting viewpoints exist regarding the optimal diet and dietary ingredients for children with diarrhea. Moreover, few studies have investigated the effect of rice soup along with ORS in the treatment of this disease. Objectives: This study aimed to explore effects of simultaneous taking of glucose oral rehydration solution (G-ORS) and rice soup in the treatment of acute diarrhea in 8 to 24-month-old children. Patients and Methods: This single-blind controlled clinical trial was conducted in the pediatric ward of 22nd of Bahman hospital, Gonabad, Iran between June 2013 and February 2014. Forty children aged 8-24 months with acute diarrhea were randomly assigned into an intervention group (G-ORS plus rice soup group) comprising 20 babies and a control group (G-ORS) of 20 children based on balanced blocking randomization. The variables under investigation were diarrhea duration, patient hospitalization, need for intravenous (IV) fluids and stool output frequency. Data was analyzed using independent samples t and chi-square test. Results: At the end of study, the time for treating acute watery diarrhea in the intervention and control groups were 21.10 ± 8.81 and 34.55 ± 5.82 hours (P < 0.001) and hospital stay were 34.05 ± 6.62 and 40.20 ± 6.32 hours (P = 0.005). Moreover, stool output frequency were 4.20 ± 0.95 and 8.00 ± 1.37 (P < 0.001) in the first 24 hours, and 2.18 ± 0.60 and 2.80 ± 0.76 (P = 0.03) in the second 24 hours of treatment in intervention and control groups, respectively. Conclusions: Rice soup regimen was highly effective and inexpensive in the treatment of acute diarrhea in children. Thus, in addition to the common treatment by G-ORS, rice soup can be consumed simultaneously with G-ORS

Effect of black surfaces to increase the average surface temperature of the earth: A global warning!

Gradual increase of the Earth's surface temperature can cause many significant changes on the climate of our planet. This problem is progressively increasing. One of the main reasons for this phenomenon is the absorption of the Sun's energy and lack of proper reflection of emissions due to man-made activities, one of which is the increase in black surfaces. In this descriptive and cross-sectional research, black surfaces in Gonabad city and the average heat absorption in amount and the increase in average heat absorption due to these surfaces were calculated by physical formula. To date, we used the information obtained from Gonabad Isallobars Office (GIO) and other related offices in the region (from 1980-2009) and performed a simple experiment to support our claim. Many factors were found to be the cause of the increase in average surface temperature of earth, such as increase in black surfaces. These surfaces appear not only on residential buildings but also on roads which have increased (637%) during this 30-year period in Gonabad city. It was calculated that 864 m3 of the air should be replaced to cool 1 m2 of the black surfaces. The average heat of the land surface in desert cities seems to increase rapidly. Therefore, it deserves much attention on the part of local managers and city officials to design plans and make some decisions for reducing or alleviating the problem as soon as possible

The attitudes of team members towards family presence during hospital-based CPR: A study based in the Muslim setting of four Iranian teaching hospitals

Background: Contrary to international guidelines recommending family presence during cardiopulmonary resuscitation (CPR), allowing family members to be present remains a matter of debate in many countries. The purpose of this study was to determine the opinions of healthcare providers from a Muslim setting concerning family-witnessed resuscitation (FWR). Methods: The sample population consisted of CPR responders in four teaching hospitals in Tehran. These centres have no policy regarding the presence of family members during resuscitation. We developed and circulated a questionnaire gathering opinions, and collated their comments. Results: From 200 respondents, 77 opposed FWR. We found that gender, age, experience, previous exposure to FWR or specialty (except for emergency physicians) did not predict opinion towards family presence during CPR. The most common reasons given for opposition to family presence were fear of psychological trauma to family members, possible interference with patient care/ decision-making, and a perceived increase in staff stress. Conclusion: In a largely Muslim community, and contrary to general guidelines, our survey suggested that the majority of CPR responders do not favour the presence of relatives during cardiopulmonary resuscitation. Any counter to this opinion would need to be based on educating team members about the possible benefits of relatives being present during resuscitation. Public education surrounding CPR would also be a fundamental element for implementing any formal programme encouraging family-witnessed CPR in hospitals such as ours. © 2010 Royal College of Physicians of Edinburgh

Algorithms for crime prediction in smart cities through data mining

The concentration of police resources in conflict zones contributes to the reduction of crime in the region and the optimization of those resources. This paper presents the use of regression techniques to predict the number of criminal acts in Colombian municipalities. To this end, a set of data was generated merging the data from the Guardia Civil with public data on the demographic structure and voting trends in the municipalities. The best regressor obtained (Random Forests) achieves a RRSE (Root Relative Squared Error) of 40.12% and opens the way to keep incorporating public data of another type with greater predictive power. In addition, M5Rules were used to interpret the results