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Journal of Clinical Laboratory Analysis 2002Considerable time and effort can be saved by simultaneously amplifying multiple sequences in a single reaction, a process referred to as multiplex polymerase chain... (Review)
Review
Considerable time and effort can be saved by simultaneously amplifying multiple sequences in a single reaction, a process referred to as multiplex polymerase chain reaction (PCR). Multiplex PCR requires that primers lead to amplification of unique regions of DNA, both in individual pairs and in combinations of many primers, under a single set of reaction conditions. In addition, methods must be available for the analysis of each individual amplification product from the mixture of all the products. Multiplex PCR is becoming a rapid and convenient screening assay in both the clinical and the research laboratory. The development of an efficient multiplex PCR usually requires strategic planning and multiple attempts to optimize reaction conditions. For a successful multiplex PCR assay, the relative concentration of the primers, concentration of the PCR buffer, balance between the magnesium chloride and deoxynucleotide concentrations, cycling temperatures, and amount of template DNA and Taq DNA polymerase are important. An optimal combination of annealing temperature and buffer concentration is essential in multiplex PCR to obtain highly specific amplification products. Magnesium chloride concentration needs only to be proportional to the amount of dNTP, while adjusting primer concentration for each target sequence is also essential. The list of various factors that can influence the reaction is by no means complete. Optimization of the parameters discussed in the present review should provide a practical approach toward resolving the common problems encountered in multiplex PCR (such as spurious amplification products, uneven or no amplification of some target sequences, and difficulties in reproducing some results). Thorough evaluation and validation of new multiplex PCR procedures is essential. The sensitivity and specificity must be thoroughly evaluated using standardized purified nucleic acids. Where available, full use should be made of external and internal quality controls, which must be rigorously applied. As the number of microbial agents detectable by PCR increases, it will become highly desirable for practical purposes to achieve simultaneous detection of multiple agents that cause similar or identical clinical syndromes and/or share similar epidemiological features.
Topics: Humans; Magnesium Chloride; Polymerase Chain Reaction
PubMed: 11835531
DOI: 10.1002/jcla.2058 -
BioTechniques Sep 1997By simultaneously amplifying more than one locus in the same reaction, multiplex PCR is becoming a rapid and convenient screening assay in both the clinical and the... (Comparative Study)
Comparative Study
By simultaneously amplifying more than one locus in the same reaction, multiplex PCR is becoming a rapid and convenient screening assay in both the clinical and the research laboratory. While numerous papers and manuals discuss in detail conditions influencing the quality of PCR in general, relatively little has been published about the important experimental factors and the common difficulties frequently encountered with multiplex PCR. We have examined various conditions of the multiplex PCR, using a large number of primer pairs. Especially important for a successful multiplex PCR assay are the relative concentrations of the primers at the various loci, the concentration of the PCR buffer, the cycling temperatures and the balance between the magnesium chloride and deoxynucleotide concentrations. Based on our experience, we propose a protocol for developing a multiplex PCR assay and suggest ways to overcome commonly encountered problems.
Topics: Buffers; DNA Primers; Dimethyl Sulfoxide; Electrophoresis, Agar Gel; Electrophoresis, Polyacrylamide Gel; Glycerol; Indicators and Reagents; Magnesium Chloride; Nucleotides; Polymerase Chain Reaction; Serum Albumin, Bovine; Solutions; Taq Polymerase; Temperature; Templates, Genetic; Time Factors
PubMed: 9298224
DOI: 10.2144/97233rr01 -
Magnesium Research Sep 2005MgSO4 is routinely used in therapeutics despite its toxicity. The aim of the present review was to compare MgSO4 and MgCl2 effects in order to answer the question... (Comparative Study)
Comparative Study Review
MgSO4 is routinely used in therapeutics despite its toxicity. The aim of the present review was to compare MgSO4 and MgCl2 effects in order to answer the question whether MgSO4 could be or not replaced by MgCl2. Considering that the two salts have both similar and proper effects, a clear-cut conclusion is not easy to draw. However, choosing MgCl2 seems advisable because of its more interesting clinical and pharmacological effects and its lower tissue toxicity as compared to MgSO4.
Topics: Animals; Female; Humans; Magnesium Chloride; Magnesium Sulfate; Obstetric Labor, Premature; Pre-Eclampsia; Pregnancy
PubMed: 16259379
DOI: No ID Found -
PloS One 2017Current treatment options for depression are limited by efficacy, cost, availability, side effects, and acceptability to patients. Several studies have looked at the... (Randomized Controlled Trial)
Randomized Controlled Trial
Current treatment options for depression are limited by efficacy, cost, availability, side effects, and acceptability to patients. Several studies have looked at the association between magnesium and depression, yet its role in symptom management is unclear. The objective of this trial was to test whether supplementation with over-the-counter magnesium chloride improves symptoms of depression. An open-label, blocked, randomized, cross-over trial was carried out in outpatient primary care clinics on 126 adults (mean age 52; 38% male) diagnosed with and currently experiencing mild-to-moderate symptoms with Patient Health Questionnaire-9 (PHQ-9) scores of 5-19. The intervention was 6 weeks of active treatment (248 mg of elemental magnesium per day) compared to 6 weeks of control (no treatment). Assessments of depression symptoms were completed at bi-weekly phone calls. The primary outcome was the net difference in the change in depression symptoms from baseline to the end of each treatment period. Secondary outcomes included changes in anxiety symptoms as well as adherence to the supplement regimen, appearance of adverse effects, and intention to use magnesium supplements in the future. Between June 2015 and May 2016, 112 participants provided analyzable data. Consumption of magnesium chloride for 6 weeks resulted in a clinically significant net improvement in PHQ-9 scores of -6.0 points (CI -7.9, -4.2; P<0.001) and net improvement in Generalized Anxiety Disorders-7 scores of -4.5 points (CI -6.6, -2.4; P<0.001). Average adherence was 83% by pill count. The supplements were well tolerated and 61% of participants reported they would use magnesium in the future. Similar effects were observed regardless of age, gender, baseline severity of depression, baseline magnesium level, or use of antidepressant treatments. Effects were observed within two weeks. Magnesium is effective for mild-to-moderate depression in adults. It works quickly and is well tolerated without the need for close monitoring for toxicity.
Topics: Adult; Aged; Antidepressive Agents; Cross-Over Studies; Depressive Disorder; Female; Humans; Magnesium Chloride; Male; Middle Aged; Treatment Outcome
PubMed: 28654669
DOI: 10.1371/journal.pone.0180067 -
Bioscience Reports Dec 2013DNA manipulation routinely requires competent bacteria that can be made using one of numerous methods. To determine the best methods, we compared four commonly used... (Comparative Study)
Comparative Study
DNA manipulation routinely requires competent bacteria that can be made using one of numerous methods. To determine the best methods, we compared four commonly used chemical methods (DMSO, MgCl2-CaCl2, CaCl2 and Hanahan's methods) on frequently used Escherichia coli (E. coli) strains: DH5α, XL-1 Blue, SCS110, JM109, TOP10 and BL21-(DE3)-PLysS. Hanahan's method was found to be most effective for DH5α, XL-1 Blue and JM109 strains (P<0.05), whilst the CaCl2 method was best for SCS110, TOP10 and BL21 strains (P<0.05). The use of SOB (super optimal broth) over LB [Luria-Bertani (broth)] growth media was found to enhance the competency of XL-1 Blue (P<0.05), dampened JM109's competency (P<0.05), and had no effect on the other strains (P>0.05). We found no significant differences between using 45 or 90 s heat shock across all the six strains (P>0.05). Through further optimization by means of concentrating the aliquots, we were able to get further increases in transformation efficiencies. Based on the optimized parameters and methods, these common laboratory E. coli strains attained high levels of TrE (transformation efficiency), thus facilitating the production of highly efficient and cost-effective competent bacteria.
Topics: Calcium Chloride; Cell Membrane Permeability; Culture Media; DNA Transformation Competence; Dimethyl Sulfoxide; Escherichia coli; Magnesium Chloride; Plasmids; Transformation, Bacterial
PubMed: 24229075
DOI: 10.1042/BSR20130098 -
Journal of the American Association For... Sep 2021Cephalopods are important in biologic and biomedical research, yet relatively little objective information is available to guide researchers and veterinarians regarding...
Cephalopods are important in biologic and biomedical research, yet relatively little objective information is available to guide researchers and veterinarians regarding the best methods for anesthetizing these animals for various experimental procedures. Recent studies demonstrate that ethyl alcohol and magnesium chloride are effective at depressing efferent and afferent neural signals in some tropical cephalopod species when measured via the pallial nerve. Here we used similar methods to test 2 temperate species ( and ) and demonstrate that (1) ethyl alcohol and magnesium chloride were effective at reversibly depressing evoked activity in the pallial nerve, (2) ethyl alcohol generally had shorter induction and recovery times compared with magnesium chloride, (3) both agents were associated with a latency between the behavioral and neural effects, and it was longer with magnesium chloride, and (4) senescent animals generally had longer induction or recovery times than young animals. Both agents successfully anesthetized both life stages; however, our data show that assessing anesthesia based solely on behavior may lead to premature commencement of invasive procedures. We conclude that temperate cephalopods can be humanely, effectively, and completely anesthetized by using these 2 agents and that the loss of neural signal we show here is consistent with true anesthesia and not merely paralysis. This relatively simple, nondestructive nerve recording technique can be applied to the study of other prospective anesthetic agents in cephalopods.
Topics: Anesthetics; Animals; Decapodiformes; Ethanol; Magnesium Chloride; Octopodiformes; Prospective Studies
PubMed: 34233805
DOI: 10.30802/AALAS-JAALAS-20-000076 -
Clinical Cardiology Nov 1993Magnesium is an essential transmembrane and intracellular modulator of the electrical activity of cardiac cells. This review provides an up-to-date consideration of the... (Review)
Review
Magnesium is an essential transmembrane and intracellular modulator of the electrical activity of cardiac cells. This review provides an up-to-date consideration of the cellular and clinical electrophysiological role of magnesium. This ubiquitous element seems to be important from both the theoretical and clinical point of view, because magnesium salts (MgSO4, MgCl2) administered intravenously are particularly effective in those arrhythmias in which the mechanism involves early or delayed after depolarization-induced triggered activity. The authors share the view that I.V. magnesium is the drug of choice in "torsade de pointes" ventricular tachycardia accompanying acquired long QT/QTU syndrome. It is complementary therapeutic agent in digitalis-induced tachycardias. Further studies are needed to elucidate magnesium's mode of action and efficacy in other types of clinical tachyarrhythmias.
Topics: Animals; Arrhythmias, Cardiac; Heart; Heart Conduction System; Humans; Magnesium; Magnesium Chloride; Magnesium Compounds; Magnesium Sulfate
PubMed: 8269653
DOI: 10.1002/clc.4960161105 -
Scientific Reports Aug 2017Mesenchymal stem cells (MSC) are osteoblasts progenitors and a variety of studies suggest that they may play an important role for the health in the field of bone...
Mesenchymal stem cells (MSC) are osteoblasts progenitors and a variety of studies suggest that they may play an important role for the health in the field of bone regeneration. Magnesium supplementation is gaining importance as adjuvant treatment to improve osteogenesis, although the mechanisms involving this process are not well understood. The objective of this study was to investigate the effects of magnesium on MSC differentiation. Here we show that in rat bone marrow MSC, magnesium chloride increases MSC proliferation in a dose-dependent manner promoting osteogenic differentiation and mineralization. These effects are reduced by 2-APB administration, an inhibitor of magnesium channel TRPM7. Of note, magnesium supplementation did not increase the canonical Wnt/β-catenin pathway, although it promoted the activation of Notch1 signaling, which was also decreased by addition of 2-APB. Electron microscopy showed higher proliferation, organization and maturation of osteoblasts in bone decellularized scaffolds after magnesium addition. In summary, our results demonstrate that magnesium chloride enhances MSC proliferation by Notch1 signaling activation and induces osteogenic differentiation, shedding light on the understanding of the role of magnesium during bone regeneration.
Topics: Animals; Bone and Bones; Boron Compounds; Cell Differentiation; Cell Proliferation; Cells, Cultured; Enzyme Inhibitors; Magnesium Chloride; Mesenchymal Stem Cells; Microscopy, Electron; Osteogenesis; Rats; Receptors, Notch; Signal Transduction; TRPM Cation Channels
PubMed: 28798480
DOI: 10.1038/s41598-017-08379-y -
The Cochrane Database of Systematic... Aug 2016The ideal intravenous fluid for kidney transplantation has not been defined, despite the common use of normal saline during the peri-operative period. The high chloride... (Meta-Analysis)
Meta-Analysis Review
BACKGROUND
The ideal intravenous fluid for kidney transplantation has not been defined, despite the common use of normal saline during the peri-operative period. The high chloride content of normal saline is associated with an increased risk of hyperchloraemic metabolic acidosis, which may in turn increase the risk of hyperkalaemia and delayed graft function. Balanced electrolyte solutions have a lower chloride content which may decrease this risk and avoid the need for dialysis due to hyperkalaemia in the immediate post-transplant period. Randomised controlled trials (RCTs) addressing this issue have used biochemical outcomes to compare fluids and have been underpowered to address patient-centred outcomes such as delayed graft function.
OBJECTIVES
To examine the effect of lower-chloride solutions versus normal saline on delayed graft function, hyperkalaemia and acid-base status in kidney transplant recipients.
SEARCH METHODS
We searched the Cochrane Kidney and Transplant's Specialised Register to 26 November 2015 through contact with the Information Specialist using search terms relevant to this review.
SELECTION CRITERIA
RCTs of kidney transplant recipients that compared peri-operative intravenous lower-chloride solutions to normal saline were included.
DATA COLLECTION AND ANALYSIS
Two independent investigators assessed studies for eligibility and risk of bias. Data from individual studies were extracted using standardised forms and pooled according to a published protocol. Summary estimates of effect were obtained using a random-effects model, and results were expressed as risk ratios (RR) and their 95% confidence intervals (CI) for dichotomous outcomes, and mean difference (MD) and 95% CI for continuous outcomes.
MAIN RESULTS
Six studies (477 participants) were included in the review. All participants were adult kidney transplant recipients and 70% of participants underwent live-donor kidney transplantation. The overall risk of bias was low for selection bias and unclear for remaining domains. There was no difference in the risk of delayed graft function (3 studies, 298 participants: RR 1.03, 95% CI 0.62 to 1.70) or hyperkalaemia (2 studies, 199 participants: RR 0.48, 95% CI 0.04 to 6.10) for participants who received balanced electrolyte solutions compared to normal saline. Intraoperative balanced electrolyte solutions compared to normal saline were associated with higher blood pH (3 studies, 193 participants: MD 0.07, 95% CI 0.05 to 0.09), higher serum bicarbonate (3 studies, 215 participants: MD 3.02 mEq/L, 95% CI 2.00 to 4.05) and lower serum chloride (3 studies, 215 participants: MD -9.93 mmol/L, 95% CI -19.96 to 0.11). There were four cases of graft loss in the normal saline group and one in the balanced electrolyte solution group, and four cases of acute rejection in the normal saline group compared to two cases in the balanced electrolyte solution group.
AUTHORS' CONCLUSIONS
Balanced electrolyte solutions are associated with less hyperchloraemic metabolic acidosis compared to normal saline, however it remains uncertain whether lower-chloride solutions lead to improved graft outcomes compared to normal saline.
Topics: Adult; Delayed Graft Function; Gluconates; Humans; Hydrogen-Ion Concentration; Hyperkalemia; Infusions, Intravenous; Isotonic Solutions; Kidney; Kidney Transplantation; Magnesium Chloride; Potassium Chloride; Ringer's Solution; Sodium Acetate; Sodium Chloride; Solutions
PubMed: 27502170
DOI: 10.1002/14651858.CD010741.pub2 -
Nutrients Jul 2018Magnesium is an essential mineral involved in a range of key biochemical pathways. Several magnesium supplements are present on the market and their degree of... (Comparative Study)
Comparative Study
Magnesium is an essential mineral involved in a range of key biochemical pathways. Several magnesium supplements are present on the market and their degree of bioavailability differs depending on the form of magnesium salt used. Aquamin-Mg is a natural source of magnesium, containing 72 additional trace minerals derived from the clean waters off the Irish coast. However, the in vitro bioaccessibility and bioavailability of Aquamin-Mg in comparison with other supplement sources of magnesium has yet to be tested. Aquamin-Mg, magnesium chloride (MgCl₂) and magnesium oxide (MgO) were subjected to gastrointestinal digestion according to the harmonized INFOGEST in vitro digestion method and in vitro bioavailability tested using the Caco-2 cell model. Magnesium concentration was measured by atomic absorption spectrophotometry (AAS). Magnesium recovery from both Aquamin-Mg and MgCl₂ was greater than for MgO. Magnesium from all three sources was transported across the epithelial monolayer with Aquamin-Mg displaying a comparable profile to the more bioavailable MgCl₂. Our data support that magnesium derived from a marine-derived multimineral product is bioavailable to a significantly greater degree than MgO and displays a similar profile to the more bioavailable MgCl₂ and may offer additional health benefits given its multimineral profile.
Topics: Caco-2 Cells; Cell Polarity; Dietary Supplements; Digestion; Enterocytes; Humans; Intestinal Absorption; Ireland; Magnesium; Magnesium Chloride; Magnesium Oxide; Minerals; Models, Biological; Nutritive Value; Osmolar Concentration; Reproducibility of Results; Spectrophotometry, Atomic
PubMed: 30018220
DOI: 10.3390/nu10070912