Developmental Pharmacogenetics of CYP2D6 in Chinese Children: Loratadine as a Substrate Drug

Objective: The elucidation of CYP2D6 developmental pharmacogenetics in children has improved, however, these findings have been largely limited to studies of Caucasian children. Given the clear differences in CYP2D6 pharmacogenetic profiles in people of different ancestries, there remains an unmet need to better understand the developmental pharmacogenetics in populations of different ancestries. We sought to use loratadine as a substrate drug to evaluate the effects of ontogeny and pharmacogenetics on the developmental pattern of CYP2D6 in Chinese pediatric patients.Methods: Chinese children receiving loratadine treatment were enrolled in the present study. The metabolite-to-parent ratio (M/P ratio), defined as the molar ratio of desloratadine to loratadine of trough concentrations samples at steady-state condition, was used as a surrogate of CYP2D6 activity. Loratadine and desloratadine were determined by LC/MS/MS method. Variants of CYP2D6 were genotyped by polymerase chain reaction for CYP2D6 *4, *10, *41 and long polymerase chain reaction for CYP2D6 *5.Results: A total of 40 patients were available for final analysis. The mean age was 4.50 (range 0.50–9.00) years and the mean weight was 19.64 (range 7.00–42.00) kg. The M/P ratio was significantly lower in intermediate metabolizers (IMs) compared to normal metabolizers (NMs) (10.18 ± 7.97 vs. 18.80 ± 15.83, p = 0.03). Weight was also found to be significantly associated with M/P ratio (p = 0.03).Conclusion: The developmental pharmacogenetics of CYP2D6 in Chinese children was evaluated using loratadine as a substrate drug. This study emphasizes the importance of evaluating the developmental pharmacogenetics in populations of different ancestries.

Download Full-text

Plasmodium spp. and Haemoproteus spp. infection in birds of the Brazilian Atlantic Forest detected by microscopy and polymerase chain reaction

Pesquisa Veterinária Brasileira ◽

10.1590/s0100-736x2015000100014 ◽

2015 ◽

Vol 35 (1) ◽

pp. 67-74 ◽

Cited By ~ 8

Author(s):

Raquel Tostes ◽

Usha Vashist ◽

Kézia K.G. Scopel ◽

Carlos L. Massard ◽

Erik Daemon ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Atlantic Forest ◽

Population Decline ◽

Wild Birds ◽

Southeastern Brazil ◽

Chain Reaction ◽

Blood Smears ◽

In Captivity ◽

The Mean ◽

Polymerase Chain

In recent years haemosporidian infection by protozoa of the genus Plasmodium and Haemoproteus, has been considered one of the most important factors related to the extinction and/or population decline of several species of birds worldwide. In Brazil, despite the large avian biodiversity, few studies have been designed to detect this infection, especially among wild birds in captivity. Thus, the objective of this study was to analyze the prevalence of Plasmodium spp. and Haemoproteus spp. infection in wild birds in captivity in the Atlantic Forest of southeastern Brazil using microscopy and the polymerase chain reaction. Blood samples of 119 different species of birds kept in captivity at IBAMA during the period of July 2011 to July 2012 were collected. The parasite density was determined based only on readings of blood smears by light microscopy. The mean prevalence of Plasmodium spp. and Haemoproteus spp. infection obtained through the microscopic examination of blood smears and PCR were similar (83.19% and 81.3%, respectively), with Caracara plancus and Saltator similis being the most parasitized. The mean parasitemia determined by the microscopic counting of evolutionary forms of Plasmodium spp. and Haemoproteus spp. was 1.51%. The results obtained from this study reinforce the importance of the handling of captive birds, especially when they will be reintroduced into the wild.

Download Full-text

The G209A mutation in the alpha-synuclein gene in Brazilian families with Parkinson's disease

Arquivos de Neuro-Psiquiatria ◽

10.1590/s0004-282x2001000500013 ◽

2001 ◽

Vol 59 (3B) ◽

pp. 722-724 ◽

Cited By ~ 8

Author(s):

Hélio A.G. Teive ◽

Salmo Raskin ◽

Fábio M. Iwamoto ◽

Francisco M.B. Germiniani ◽

Maria H.H. Baran ◽

...

Keyword(s):

Parkinson’S Disease ◽

Polymerase Chain Reaction ◽

Parkinson's Disease ◽

Family History ◽

Restriction Enzyme ◽

Alpha Synuclein ◽

Chain Reaction ◽

The Mean ◽

Polymerase Chain ◽

Mutation Assay

A missense G209A mutation of the alpha-synuclein gene was recently described in a large Contursi kindred with Parkinson's disease (PD). The objective of this study is to determine if the mutation G209A of the alpha-synuclein gene was present in 10 Brazilian families with PD. PD patients were recruited from movement disorders clinics of Brazil. A family history with two or more affected in relatives was the inclusion criterion for this study. The alpha-synuclein G209A mutation assay was made using polymerase chain reaction and the restriction enzyme Tsp45I. Ten patients from 10 unrelated families were studied. The mean age of PD onset was 42.7 years old. We did not find the G209A mutation in our 10 families with PD. Our results suggest that alpha-synuclein mutation G209A is uncommon in Brazilian PD families.

Download Full-text

Molecular tracking of pathogens in central venous catheter

The Journal of Vascular Access ◽

10.1177/1129729820934354 ◽

2020 ◽

pp. 112972982093435

Author(s):

Maristela Oliveira Lara ◽

Thabata Coaglio Lucas ◽

Evanguedes kalapothakis ◽

Ronaldo Luis Thomasini ◽

Carla Jorge Machado

Keyword(s):

Polymerase Chain Reaction ◽

Central Venous Catheter ◽

Venous Catheter ◽

Central Venous ◽

Chain Reaction ◽

Microbiological Methods ◽

Catheter Tip ◽

The Mean ◽

Polymerase Chain ◽

Molecular Tracking

Background: Central venous catheter–related bloodstream infection is an important adverse event in health care. Molecular methods are not yet substitutive of microbiological in the detection of the pathogens responsible for the infection, but they can help in the epidemiological characterization. Aim: To detect bacteria by polymerase chain reaction, from material extracted from the tip of central catheters of patients suspected of infection at the intensive care unit. Methods: Catheters (n = 34) of patients suspected of central venous catheter–related infection were analyzed by polymerase chain reaction. The findings were compared with culture of catheter tip and blood cultures performed by the hospital. Findings: The prevalence of bacteria was Staphylococcus aureus (50%), Enterococcus faecalis (41.2%), Klebsiella pneumoniae (32.4), Pseudomonas aeruginosa (20.6%), Acinetobacter baumannii (38.2%), Escherichia coli (2.9%), and Enterobacter cloacae (0%). No blood culture showed bacterial growth, the culture of catheter tip revealed bacteria in 21 (61.8%) and the polymerase chain reaction had positivity in 31 (91.2%) of the catheters. The mean central venous catheter time was 11 days, and the jugular vein was the site of insertion. Conclusion: The molecular method identified more bacteria than microbiological methods and revealed colonization of the catheters. The most commonly found bacteria are in the environment and in the microbiota of the skin, which suggests contamination by the hands of health professionals and points out the need for more efforts in preventive strategies.

Download Full-text

NR3C1 polymorphisms in Brazilians of Caucasian, African, and Asian ancestry: glucocorticoid sensitivity and genotype association

Arquivos Brasileiros de Endocrinologia & Metabologia ◽

10.1590/0004-2730000002868 ◽

2014 ◽

Vol 58 (1) ◽

pp. 53-61 ◽

Cited By ~ 12

Author(s):

Manoel Carlos L. A. Souza ◽

Clarissa S. Martins ◽

Ivan M. Silva Junior ◽

Rosangela S. Chriguer ◽

Ana C. Bueno ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Cell Proliferation ◽

Healthy Population ◽

Chain Reaction ◽

Allelic Discrimination ◽

Asian Ancestry ◽

Hardy Weinberg Equilibrium ◽

The Mean ◽

Polymerase Chain ◽

Pituitary Adrenal Axis

Objective : The Brazilian population has heterogeneous ethnicity. No previous study evaluated NR3C1 polymorphisms in a Brazilian healthy population. Materials and methods : We assessed NR3C1 polymorphisms in Brazilians of Caucasian, African and Asian ancestry (n = 380). In a subgroup (n = 40), we compared the genotypes to glucocorticoid (GC) sensitivity, which was previously evaluated by plasma (PF) and salivary (SF) cortisol after dexamethasone (DEX) suppression tests, GC receptor binding affinity (K d ), and DEX-50% inhibition (IC 50 ) of concanavalin-A-stimulated mononuclear cell proliferation. p.N363S (rs6195), p.ER22/23EK (rs6189-6190), and BclI (rs41423247) allelic discrimination was performed by Real-Time PCR (Polymerase Chain Reaction). Exons 3 to 9 and exon/intron boundaries were amplified by PCR and sequenced. Results : Genotypic frequencies (%) were: rs6195 (n = 380; AA:96.6/AG:3.14/GG:0.26), rs6189-6190 (n = 264; GG:99.6/GA:0.4), rs41423247 (n = 264; CC:57.9/CG:34.1/GG:8.0), rs6188 (n = 155; GG:69.6/GT:25.7/TT:4.7), rs258751 (n = 150; CC:88.0/CT:10.7/TT:1.3), rs6196 (n = 176; TT:77.2/TC:20.4/CC:2.4), rs67300719 (n = 137; CC:99.3/CT:0.7), and rs72542757 (n = 137; CC:99.3/CG:0.7). The rs67300719 and rs72542757 were found only in Asian descendants, in whom p.N363S and p.ER22/23EK were absent. The p.ER22/23EK was observed exclusively in Caucasian descendants. Hardy-Weinberg equilibrium was observed, except in the Asian for rs6188 and rs258751, and in the African for p.N363S. The K d , IC 50 , baseline and after DEX PF or SF did not differ between genotype groups. However, the mean DEX dose that suppressed PF or SF differed among the BclI genotypes (P = 0.03). DEX dose was higher in GG- (0.7 ± 0.2 mg) compared to GC- (0.47 ± 0.2 mg) and CC-carriers (0.47 ± 0.1 mg). Conclusion : The genotypic frequencies of NR3C1 polymorphisms in Brazilians are similar to worldwide populations. Additionally, the BclI polymorphism was associated with altered pituitary-adrenal axis GC sensitivity.

Download Full-text

Influenza species and subtypes circulation among hospitalized patients in Laleh hospital during two influenza seasonal (2016-2017 and 2017-2018) using a multiplex Real Time-Polymerase Chain Reaction

Infectious Disease Reports ◽

10.4081/idr.2020.8139 ◽

2020 ◽

Vol 12 (1) ◽

Author(s):

Iman Rezaee Azhar ◽

Minoo Mohraz ◽

Masoud Mardani ◽

Mohammad Ali Tavakoli ◽

Amin Ehteshami Afshar ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Real Time ◽

Real Time Pcr ◽

Simultaneous Detection ◽

Hospitalized Patients ◽

P Value ◽

Chain Reaction ◽

Short Period ◽

The Mean ◽

Polymerase Chain

The introduction of polymerase chain reaction (PCR) techniques has improved the detection of respiratory viruses, particularly with the use of multiplex real-time technique with the capability of simultaneous detection of various pathogens in a single reaction. The aim of this study was to apply the above technology for the diagnosis of influenza infections and at the same time to differentiate between common flu species between hospitalized patients in Laleh hospital (Iran) between two flu seasons (2016- 2017 and 2017-2018). Different respiratory specimens were collected from 540 patients from a period of December 2016 to May 2018 and were sent to the laboratory for molecular diagnosis. RNAs were extracted and subsequently, a multiplex real time PCR identifying flu A, flu B and typing flu A (H1N1) was carried out. The mean age of patients was 47.54±23.96. 216 (40%) and 321 (60%) of subjects were male and female, respectively. 219 out of 540 (40.5%) were positive for influenza infection including flu A (n=97, 44.3%), flu A (H1N1) (n=45, 20.7%) and flu B (n=77, 35%). Flu A was the dominant species on 2016-2017 and flu B was the major species on 2017-2018. Flu A (H1N1) was comparable in both time periods. Flu infections were most frequently diagnosed in age groups 21-40. Flu-positive patients suffered more from body pain and sore throat than flunegative patients with significant statistical difference (P values <0.001). The mean duration of hospitalization was shorter for flu-positive patients (P value = 0.016). Application of multiplex real time PCR could facilitate the influenza diagnosis in a short period of time, benefiting patients from exclusion of bacterial infections and avoiding unnecessary antibiotic therapy. Influenza diagnosis was not achieved in up to 60% of flu-like respiratory infections, suggesting the potential benefit of adopting the same methodology for assessing the involvement of other viral or/and bacterial pathogens in those patients.

Download Full-text

Utilization of herpes simplex PCR assays for cerebrospinal fluid in a pediatric health care setting

Canadian Journal of Microbiology ◽

10.1139/w01-028 ◽

2001 ◽

Vol 47 (5) ◽

pp. 392-396 ◽

Cited By ~ 2

Author(s):

Nevio Cimolai ◽

Eva E Thomas ◽

Rusung Tan ◽

Alan Hill

Keyword(s):

Polymerase Chain Reaction ◽

Cerebrospinal Fluid ◽

Herpes Simplex ◽

Chain Reaction ◽

Herpes Simplex Viruses ◽

Pediatric Health Care ◽

The Mean ◽

Polymerase Chain ◽

Pcr Assays ◽

To Receive

An assessment was made of the utilization and impact of a diagnostic polymerase chain reaction (PCR) assay for the diagnosis of herpes simplex viruses (HSV) 1 and 2 in cerebrospinal fluid of children who attended a Canadian pediatric referral centre. One hundred and three assays were performed on specimens from 103 patients during the period August 1997 to September 1998. Patient ages ranged from newborn to 16 years. Indications for HSV PCR included seizures with or without fever (56.3%), aseptic meningitis (16.5%), and encephalopathy with or without fever (10.7%). Only 2 of 103 (1.9%) assays were positive, including one each for HSV1 and HSV2. Control specimens that were seeded with virus indicated inhibition for 24.3, 8.8, and 6.8% of assays for HSV1, HSV2, and both HSV1 and HSV2, respectively. The mean turn-around time for HSV PCR was 2.5 days, and 90.3% were completed in less than 5 days. Acyclovir was administered to 78.6% of the patients overall; the results of the HSV PCR impacted on the treatment courses for 36 individuals. Nevertheless, 16.5% of patients continued to receive extended courses of antiviral therapy despite negative HSV PCR assays. Although it is desirable to decrease the frequency of PCR inhibitions and to further decrease the interval to assay completion, HSV PCR does have a significant impact on antiviral use in this setting.Key words: herpes simplex, polymerase chain reaction, central nervous system.

Download Full-text

Selection of male goat embryos in vitro by using swim-up of epididymal spermatozoa

The Iraqi Journal of Veterinary Medicine ◽

10.30539/iraqijvm.v42i1.35 ◽

2018 ◽

Vol 42 (1) ◽

pp. 79-86

Author(s):

Ihsan H. S. Al-Timimi

Keyword(s):

Polymerase Chain Reaction ◽

In Vitro Fertilization ◽

In Vitro Maturation ◽

Chain Reaction ◽

Vitro Fertilization ◽

The Mean ◽

Polymerase Chain ◽

Epididymal Spermatozoa ◽

Selection Of

The main objectives of this study is the separation of X from Y bearing epididymal spermatozoa of local buck by swim-up, and the use of this spermatozoa for in vitro fertilization to determine the percentage of produced male and female embryos. The sex of produced embryo was identified by polymerase chain reaction. Testis of the local buck were obtained from Al-Shu'alah abattoir and the epididymal spermatozoa were harvested from the cauda by and submitted to in vitro maturation prior to separation of X from Y bearing spermatozoa and prior to their use for in vitro fertilization. For the separation of epididymal spermatozoa, swim-up technique was used with centrifugation at 200×g or 300×g. The centrifugation at 200×g showed that 41.84±1.39 % of spermatozoa were detected in the supernatant while the precipitate contained 50.69±0.71 and the mean of the sperm lost was 7.65±0.93. After centrifugation, spermatozoa in the supernatant were used for in vitro fertilization of matured oocytes. The sex of in vitro produced goat embryos was determined by polymerase chain reaction using specific primers to detect of SRY gene. The percentage of total goat embryos obtained after in vitro fertilization by sperms selected using swim-up at centrifugation force of 200×g recorded 79.66 % male embryos while female embryos recorded only 20.33 %. At the end, the results showed the ability of selection male embryos in caprine by application of swim-up technique on epididymal spermatozoa with centrifugation at 200×g.

Download Full-text

Virus Load and Incidence Olfactory, Gustatory, Respiratory, Gastrointestinal Disorders in COVID-19 Patients: A Retrospective Cohort Study

10.22541/au.161635941.12859469/v1 ◽

2021 ◽

Author(s):

Mohammad Hosein Taziki Balajelini ◽

Abdolhalim Rajabi ◽

Masoud Mohammadi ◽

Hadi Razavi Nikoo ◽

Alijan Tabarraei ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Cohort Study ◽

Viral Load ◽

Retrospective Cohort Study ◽

Retrospective Cohort ◽

Gastrointestinal Symptoms ◽

Chain Reaction ◽

Ct Value ◽

The Mean ◽

Polymerase Chain

Objectives: This study investigated the relationship between viral load and the incidence and recovery of olfactory and gustatory dysfunction (OD and GD), incidence of respiratory and gastrointestinal symptoms in COVID-19 patients. Design: A Retrospective Cohort Study. Setting and Participants: In total 599 outpatients’ cases polymerase chain reaction (PCR)–confirmed COVID-19-positive patients in Golestan province were included in the study. Main Outcome Measures: The incidence of OD, GD, their severity and the time of recovery was determined. The association of these variables with cycle threshold (CT) values of SARS-CoV-2 polymerase chain reaction was assessed. Results: The mean age of patients was 38.27 ± 13.62 years. The incidence of general symptoms included myalgia 70.1%, headache 51.8%, fever 47.7%, and dyspnea 21.4%. 41.9% of patients had gastrointestinal symptoms, including abdominal pain 26.5%, diarrhea 25.2%, nausea 20.5%, and vomiting 12.9%. 12.2% of patients had comorbidity. The trimester recovery rates of OD and GD were 93.94% and 94.74%, respectively. The mean recovery time of OD and GD was 14.56±13.37 and 13.8±3.77 days, respectively. The mean CT value in all patients was 27.45±4.55. There were significant associations between CT value with headache (P=0.04), GD (P=0.002) and OD (P=0.001). Conclusions: The intensity and the recovery of OD and GD in Covid-19 patients may be affected by initial viral load. Unlike to respiratory and gastrointestinal symptoms, the OD and GD were associated with lower viral load. Therefore, it may be recommended to use these clinical symptoms as an indicator in the initial screening of patients during pandemics.

Download Full-text

Covid-19 in outpatients—Is fever a useful indicator for SARS-CoV-2 infection?

PLoS ONE ◽

10.1371/journal.pone.0246312 ◽

2021 ◽

Vol 16 (2) ◽

pp. e0246312

Author(s):

Anne Schneider ◽

Holger Kirsten ◽

Franziska Lordick ◽

Florian Lordick ◽

Christoph Lübbert ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Body Temperature ◽

Final Analysis ◽

University Hospital ◽

Rt Pcr ◽

Chain Reaction ◽

Course Of Illness ◽

Oropharyngeal Swab ◽

Polymerase Chain ◽

Telephone Calls

Objective Understanding mild to moderate symptoms of coronavirus disease 2019 (Covid-19) is important in order to identify active cases early and thus counteract transmission. Methods In March 2020, Leipzig University Hospital established an outpatient clinic for patients potentially infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Confirmed cases with mild to moderate symptoms self-isolated at home and were followed-up by daily telephone calls for at least 14 days. Symptoms and course of illness of these patients are reported here. Results From March 20 to April 17, 2020, 1460 individuals were tested for SARS-CoV-2 by naso- or oropharyngeal swab for real-time polymerase chain reaction (RT-PCR). Covid-19 was confirmed in 91 (6.2%) patients, of which 87 were included in the final analysis. Patients presented for testing after a mean of 5.9 days (IQR = 2.0–8.5). The median age was 37.0 years (IQR = 28.5–53), and 48 (55.2%) were female. Five (5.7%) patients required hospital admission during the course of illness. Most frequently reported symptoms were fatigue (n = 64, 74%), cough (n = 58, 67%), and hyposmia/hypogeusia (n = 44, 51%). In contrast to previous reports, fever occurred in less than a third of patients (n = 25, 29%). By day 14, more than half of the patients had recovered completely (n = 37/70, 52.9%). Conclusions Fever seems to be less common in patients of relatively young age diagnosed with mild to moderate Covid-19. This suggests that body temperature alone may be an insufficient indicator of SARS-CoV-2 infection.

Download Full-text

MORPHOLOGICAL AND MOLECULAR IDENTIFICATION OF ZOONOTIC TREMATODE METACERCARIAE (Haplorchis taitui) IN FRESHWATER FISH IN IRAN

Veterinária e Zootecnia ◽

10.35172/rvz.2021.v28.442 ◽

2021 ◽

Vol 28 ◽

pp. 1-9

Author(s):

Alireza Golchin Manshadi

Keyword(s):

Public Health ◽

Polymerase Chain Reaction ◽

Polymerase Chain Reaction Product ◽

Deoxyribonucleic Acid ◽

Nucleotide Sequence Analysis ◽

Chain Reaction ◽

Mean Intensity ◽

The Mean ◽

Polymerase Chain ◽

Haplorchis Taichui

The intestinal trematode, Haplorchis taichui (Nishigori, 1924), is an important in public health that causes infection in humans and animals especially in Asia and in parts of Africa and the Americas. Haplorchis taitui metacercariae were found in the gills of Cyprinion macrostomus (Heckel) and Capoeta barroisi persica (Karaman) collected from the Shapour River. Morphological excysted metacercariae were identified as wet mounts under a stereomicroscope. Then, the samples were subjected to molecular analysis. The result showed that 69% of examined fish (n = 30) were diagnosed infected with encysted metacercariae in gills. The mean intensity was 8.3 ± 16.9 parasites per fish. The morphometrical values agree with the findings of other studies with the small differences and polymerase chain reaction product length and nucleotide sequence analysis of 18S ribosomal deoxyribonucleic acid gene showed a similarity of over 99% between the specimens and the Haplorchis taichui (Nishigori, 1924) recorded in GenBank.

Download Full-text