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Theophylline

Theophylline is a methylxanthine derivative with bronchodilator and respiratory stimulant properties.
It is commonly used in the treatment of asthma, chronic obstructive pulmonary disease (COPD), and other respiratory conditions.
Theophylline works by relaxing the muscles of the airways, improving airflow, and reducing inflammation.
It may also have effects on the central nervous system and cardiovascular system.
Proper dosing and monitoring are essential, as theophylline has a narrow therapeutic window and can interact with other medications.
Researchers can optimize their theophylline studies using PubCompare.ai, an AI-driven platform that compares research protocols to identify the most reproducible and accurate procedures from the literature, preprints, and patents.
This intuitive tool streamlines protocol selection and comparison, enhancing the efficiency and quality of theophylline research.

Most cited protocols related to «Theophylline»

All riboswitch and promoter sequences are listed in Supplementary Data. Theophylline, fluoride and DNT riboswitches were constructed and inserted into an mRFP1 fluorescent protein expression vector, derived from plasmid pFTV1 (ColE1 origin, CmR) (31 (link)). Three theophylline riboswitches (Theo-40, Theo-41 and Theo-45) also controlled the translation of a fusion mRFP1 protein. To create the fusion protein, four or five non-rare codons were introduced between the start codon and SacI restriction site within mRFP1 coding section. All the riboswitches were constructed using standard molecular cloning. Briefly, DNA fragments were computationally designed, synthesized and assembled using either annealing of oligonucleotides, polymerase chain reaction (PCR) assembly of oligonucleotides, or PCR amplification of gBLOCK DNA fragments (Integrated DNA Technologies). DNA fragments were then digested by XbaI and SacI restriction enzymes, followed by ligation with digested plasmid, transformation, plating on selective media and verification of purified plasmid by sequencing. Similarly, promoter replacements were performed by annealing designed pairs of oligonucleotides, followed by digestion with BamHI/XbaI restriction enzymes, ligation, transformation, selective plating and verification by sequencing.
The promoters AEB-3, J23100 and LmrA were selected or designed to significantly vary riboswitch transcription rates (Supplementary Figure S17). The promoter AEB-3 is a result of mutating the −10 and −35 hexamers of promoter J23100, resulting in 10-fold lower transcription rate. The promoter LmrA is a near-consensus promoter with a 5-fold higher transcription rate. Unless noted otherwise in the text, all theophylline and fluoride riboswitches use the J23100 promoter, while all DNT riboswitches used the AEB-3 promoter.
Theophylline and TMR riboswitches were then constructed in plasmids expressing the luciferase reporter protein using standard molecular cloning. The plasmid is derived from the pBESTluc vector (Promega) initially using a pUC19 origin, where the riboswitch-reporter mRNA is transcribed by a Ptac promoter. As described in the text, plasmid origins were replaced with either pBAC, p15A or pFTV1 by PCR amplifying the expression cassette (promoter to transcriptional terminator) and digesting it with BamHI and SpeI, followed by ligation to the corresponding digested vectors, transformation, selective plating and verification by sequencing.
Dopamine and thyroxine riboswitches were constructed in pFTV1-derived plasmids containing the luciferase expression cassette, where pFTV1 was previously modified to insert AatII and HindIII restriction sites after the Ptac promoter and after the start codon, respectively. Riboswitch-encoding DNA fragments were PCR-amplified from designed gBLOCKs, followed by digestion with AatII and HindIII, ligation to digested plasmids, transformation, selective plating and verification by sequencing.
Publication 2015
6-(3-propylthio-1,2,5-thiadiazol-4-yl)-1-azabicyclo(3.2.1)octane Cloning Vectors Codon Codon, Initiator Digestion DNA-Directed DNA Polymerase DNA Restriction Enzymes Dopamine Fluorides Ligation Luciferases Mrfp1 protein Oligonucleotides Plasmids Polymerase Chain Reaction Promega Protein Biosynthesis Proteins Riboswitch RNA, Messenger Surgical Replantation Theophylline Thyroxine Transcription, Genetic
Cultures with candidate drift plasmids contained: pT7 (pJC173b), pTet (pJC148i2), psp (pJC175e), psp-tet (pJC175g/h/k/m/n), psp-tet on MP (pJC184d5), psp-tet on colE1-spect plasmid (pJC184d-Sp). Cultures were grown to mid-log phase in Davis rich media and infected with an excess of SP-T7 for 10 min at 37°C. Cells were pelleted and washed to remove residual excess phage. Cells were re-inoculated into fresh Davis rich media and grown to OD600 ∼0.8, pelleted, and supernatants were saved for titering by plaque assay on S1030 E. coli cells carrying a PT7-gene III AP (pJC173b).. OD600 values were measured to normalize phage titers.
Cultures with candidate negative selection plasmids contained PTet-gene III (pJC148i2) and either Plac-C-domain (pJC156a2), Plac-C83 (pJC156c2), Plac-N1-N2-C83 (pJC156j2), Plac-N2-C83 (pJC156m2), or Plac-N1*-N2-C83 (pJC156o2). Cells were treated with SP-T7 phage as described above and then cultured in 2xYT media with either 4 ng/mL or 20 ng/mL ATc and in the presence or absence of 2 mM IPTG. Phage titers were assayed as described above.
Cultures for theophylline-dependent negative selection contained PT7-gene III (pJC173c) and PT3-theophylline riboswitch-gene III-neg (pJC174c-R5). Cells were treated with either SP-T7192.2 or SP-T748.3 and then cultured with varying concentrations of theophylline. Phage titers were assayed as described above.
Publication 2014
5'-palmitoyl cytarabine Bacteriophages Cells Culture Media Dental Plaque Escherichia coli Genes Isopropyl Thiogalactoside Plasmids Riboswitch Theophylline Tomography, Emission-Computed, Single-Photon
In this genome-wide association study, we used a two-stage design to identify novel and significant genome-wide associations that confer susceptibility to moderate-to-severe asthma. We used a two-stage case-control design, with variants that showed suggestive association (p<1 × 10−6) in stage 1 tested in stage 2 and then meta-analysed across the two stages to maximise power.
For stage 1, we selected individuals of European ancestry with moderate-to-severe asthma who had been recruited from primary and secondary care settings across the UK as part of the Genetics of Asthma Severity and Phenotypes (GASP) initiative, with additional cases included from the U-BIOPRED asthma cohort26 (link) and the UK Biobank May, 2015,27 , 28 (link) genetic data release (appendix). Genotyped data were assessed for quality control (details are in the appendix). From GASP and U-BIOPRED, we identified patients with moderate-to-severe asthma by assessing clinical records that indicated that a patient was taking medication required for patients defined as having moderate-to-severe asthma according to the British Thoracic Society (BTS) 2014 guidelines.29 From the UK Biobank, cases of moderate-to-severe asthma were defined as having asthma diagnosed by a doctor, taking medication for asthma, no diagnosis of emphysema or chronic bronchitis by a doctor, and meeting the definition of moderate-to-severe asthma by BTS criteria. Therefore, cases were selected from individuals for whom medication information was available and who met BTS stage 3–5 criteria—ie, for stage 3, taking a long-acting β2 agonist plus inhaled corticosteroid; stage 4, taking higher dose inhaled corticosteroids than stage 3 patients, and addition of a fourth drug (eg, leukotriene receptor antagonist, theophylline); and stage 5, taking oral corticosteroid or omalizumab, or both. A complete list of medications used to identify patients with moderate-to-severe asthma is in the appendix. Controls for stage 1 were identified from the UK Biobank by taking the remaining subjects for whom genotyped data were available that passed quality control and excluding individuals with asthma, rhinitis, eczema, allergy, emphysema, or chronic bronchitis as diagnosed by a doctor, or if medication data were not available to assign to either the mild-moderate or moderate-severe asthma group. Additional controls for stage 1 were included from U-BIOPRED to ensure we had controls from each cohort. Patients in the U-BIOPRED cohort had not been screened for rhinitis or eczema, and so this information was not available for these controls at time of selection. For stage 2, both cases and controls were selected from the UK Biobank May, 2017, release using the same criteria to define cases and controls as in stage 1. There was no overlap in the patients included in stage 1 and stage 2. A case-control ratio of 1:5 was chosen for both stages to balance power and computational time. Cases and controls were matched across age and sex strata, and in stage 1 across genotyping arrays. All cohorts included individuals with self-reported European ancestry; individuals of non-European ancestry were excluded to reduce confounding of the study by ancestry.
UK Biobank has ethical approval from the UK National Health Service (NHS) National Research Ethics Service (Ref 11/NW/0382). All other studies were approved by an appropriate ethics committee. Informed consent was obtained from all participants.
Publication 2019
Adrenal Cortex Hormones Asthma Bronchitis, Chronic Eczema Emphysema Ethics Committees Europeans Genome Genome-Wide Association Study Hypersensitivity Leukotriene Antagonists Omalizumab Patients Pharmaceutical Preparations Phenotype Physicians Reproduction Rhinitis Secondary Care Susceptibility, Disease Theophylline
Murine embryonic stem cells were differentiated into embryoid bodies using the hanging-drop method, with cardiac differentiation monitored by epifluorescence using α-actinin antibody (1:1,000) and live microscopy (34 (link), 48 (link)). RNA was processed by quantitative RT-PCR using the Light Cycler (Roche) and QuantiTect SYBR Green kit (QIAGEN) to quantify Nkx2.5 (forward, reverse primers: 5′-TGCAGAAGGCAGTGGAGCTGGACAAGCC-3′ and 5′-TTGCACTTGTAGCGACGGTTCTGGAACCA-3′), MEF-2C (5′-AGATACCCACAACACACCACGCGCC-3′ and 5′-ATCCTTCAGAGACTCGCATGCGCTT-3′), GATA-4 (5′-GGAATTCAAGATGAACGGCATCAAC-3′ and 5′-TGAATTCTCAACCTGCTGGCGTCTTAGA-3′), and β-MHC (5′-GCCAAAACACCAACCTGTCCAAGTTC-3 and 5′-CTGCTGGAGAGGTTATTCCTCG-3′) mRNA expression normalized to β-tubulin. From day 7 embryoid bodies, cardiopoietic progenitor cells were isolated by Percoll purification and visualized through laser confocal examination using MEF-2C (1:400; Cell Signaling Technologies), Nkx2.5 (1:300; Santa Cruz Biotechnology, Inc.), GATA-4 (1:300, Santa Cruz Biotechnology; Inc.), and α-actinin (1:1,000; Sigma-Aldrich). In a subset of experiments, the endodermal layer was eliminated from embryoid bodies through generation of hanging drops in the presence of recombinant leukemia inhibitory factor (10 U/μl) once differentiation has been initiated (38 (link)). Isolated visceral endoderm-like cells were derived from an F9 cell population (American Type Culture Collection) with retinoic acid (1 μM), dibutynyl cAMP (0.5 mM), and theophylline (0.5 mM), with phenotype confirmed through comparison with END-2 cells (67 (link)). Conditioned medium obtained after 24 h of culture was used for dissection of procardiogenic signaling by proteomic analysis. To stimulate cardiopoiesis of embryonic stem cells cultured in monolayer at 100 cells/cm2, cells were stimulated with recombinant TGF-β1 (2.5 ng/ml), BMP-2 and -4 (5 ng/ml), activin-A (5 ng/ml), FGF-2 and -4 (10 ng/ml), IL-6 (100 ng/ml), IGF-1 and -2 (50 ng/ml), VEGF-A (10 ng/ml), and EGF (2.5 ng/ml) either in singular or combinatorial fashion with cellular response monitored by confocal microscopy. When recruited from a monolayer of embryonic stem cells, the cardiopoietic population was enriched using a dual interface Percoll gradient to separate sarcomere-rich high density cardiomyocytes (34 (link)) from the lower density sarcomere-poor cardiopoietic phenotype. Cardiopoietic cell proliferation and purity was assessed by ArrayScan multichannel fluorescence automated microscopy (Cellomics) using MEF-2C and α-actinin antibodies, along with DAPI nuclear staining. Cell morphology was resolved by field emission scanning or transmission electron microscopy. Action potentials and voltage–current relationships were acquired by patch-clamp electrophysiology. Calcium dynamics were tracked in Fluo 4-AM–loaded cells using laser confocal line scanning (48 (link)).
Publication 2007
Actinin Action Potentials activin A Antibodies Bone Morphogenetic Protein 2 Calcium Cell Proliferation Cells Culture Media, Conditioned DAPI Dissection Embryoid Bodies Embryonic Stem Cells Endoderm Fibroblast Growth Factor 2 Fluo 4 Fluorescence Heart IGF1 protein, human Immunoglobulins LIF protein, human Light Maritally Unattached Microscopy Microscopy, Confocal Mus Myocytes, Cardiac Oligonucleotide Primers Percoll Phenotype Reverse Transcriptase Polymerase Chain Reaction RNA, Messenger Sarcomeres Stem Cells SYBR Green I TGF-beta1 Theophylline Transmission Electron Microscopy Tretinoin Tubulin Vascular Endothelial Growth Factors
These were multinational, replicate, phase III, multicentre, randomised, double-blind, active-controlled, five-arm, parallel-group studies, registered with ClinicalTrials.gov (Study 1237.5: NCT01431274; Study 1237.6: NCT01431287) (fig. 1). Three primary end points were evaluated after 24 weeks of treatment: forced expiratory volume in 1 s (FEV1) area under the curve from 0 to 3 h (AUC0–3) response (in each individual trial), trough FEV1 response in each individual trial (response defined as change from baseline; mean of the values of 1 h and 10 min prior to the first dose of study medication); and St George's Respiratory Questionnaire (SGRQ) total score (SGRQ was analysed in a pre-specified combined analysis of data from both studies). Pulmonary function tests (PFTs) were performed on day 1 and at weeks 2, 6, 12, 18, 24, 32, 40 and 52. SGRQ was completed on day 1 and after 12, 24 and 52 weeks, prior to PFTs and all other procedures. Details of the study design, assessments performed and statistical methodology are provided in table S1 of the online supplementary material.

Study design (Study 1237.5: NCT01431274; Study 1237.6: NCT0143 1287). R: randomisation; FDC: fixed- dose combination. #: primary end-point assessment.

Patients continued to receive treatment with inhaled corticosteroids as required and were provided with salbutamol/albuterol metered-dose inhaler (100 μg per actuation) as rescue medication to be used as necessary at any point during the trial. Temporary increases in the dose or addition of oral steroids or theophylline preparations were allowed during the treatment portion of the study; PFTs were not performed within 7 days of the last administered dose.
Publication 2015
Adrenal Cortex Hormones Albuterol DNA Replication Inhalation Therapy Metered Dose Inhaler Patients Pharmaceutical Preparations Respiratory Rate Steroids Tests, Pulmonary Function Theophylline Volumes, Forced Expiratory

Most recents protocols related to «Theophylline»

This was a quasi-experimental study with a pre-post design that analyzes the effect of Vitamin D analog supplementation in women with uterine prolapse. We included all postmenopausal women diagnosed with grade III and IV uterine prolapse who came to outpatient clinic of Dr. Hasan Sadikin Bandung from August 2021 to November 2021. Uterine prolapse diagnosis and staging were based on Pelvic Organ Prolapse Quantification (POP-Q) system. Exclusion criteria were as follows:

Patients with comorbidities, such as chronic cough and chronic constipation (these symptoms last for a minimum of 8 weeks).

Patients diagnosed with diseases related to Vitamin D metabolism disorders such as: diabetes mellitus, chronic kidney failure, or malignant diseases

Those who had a history of gastrectomy or jejunoileostomy surgery.

Patients who are currently or have a history of taking cholesterol-lowering drugs (statins and fibrates), anticonvulsants, thiazides, theophylline, orlistat, cimetidine, and Vitamin D supplementation one month prior to this study.

Patients withdrawing from our research.

During the initial presentation, we collected the following data: age, parity, body mass index (BMI), hemoglobin levels, and calcium levels. All subjects were then given 0.5 mcg of Vitamin D analog supplementation for 3 months. We also collected and compared the following data before and after Vitamin D analog supplementation: (1) Vitamin D and VDR serum levels, (2) Levator ani muscle strength, and (3) Hand grip muscle strength. Levator ani muscle strength was measured using perineometer, while handgrip muscle strength was evaluated using hand grip dynamometer.
We tabulated all patients' data on a customized spreadsheet and performed data analysis on Statistical Produce and Service Solutions SPSS software version 25 for Windows (IBM Corp, Armonk, New York, USA). Descriptive statistics were performed as appropriate. Analytical statistics were performed using t test or Wilcoxon test as required, with p < 0.05 considered as significant.
Written informed consent was provided to all study participants prior to engaging in any study-related procedures. Ethical approval of this study was granted by the Health Research Ethics Committee of Hasan Sadikin Hospital, Bandung under the following registration number: LB.02.01/X.6.5/213/2021. This study was conducted according to Declaration of Helsinki. All research procedures were performed in accordance with relevant guidelines and regulations.
Publication 2023
Anticholesteremic Agents Anticonvulsants Calcium, Dietary Cimetidine Constipation Cough Diabetes Mellitus Ergocalciferol Ethics Committees, Clinical Fibrates Gastrectomy Grasp Hemoglobin Hydroxymethylglutaryl-CoA Reductase Inhibitors Index, Body Mass Kidney Failure, Chronic Metabolic Diseases Muscle Strength Muscle Tissue Operative Surgical Procedures Orlistat Patients Pelvic Organ Prolapse Serum Theophylline Thiazides Uterine Prolapse Woman
HEK293T/17 cells (ATCC, RRID: CVCL_1926) were stably transfected with vectors encoding GFP (for comparative purposes only), ADCY5 wild-type (ADCY5wt), and ADCY5 R418W mutant (ADCY5mut). Cells were cultured in Dulbecco’s modified Eagle’s Medium (DMEM) supplemented with 10% (v/v) fetal bovine serum (FBS) for two days in six-well plates at 37°C and 5% CO2; 4.4 x 105 cells were cultivated in one well. Each treatment was performed in triplicate. Cells were incubated with caffeine, theophylline, and istradefylline at 37°C and 5% CO2 using different concentrations (caffeine and theophylline: 1 μM, 10 μM, 100 μM, 1 mM; istradefylline: 1 nM, 10 nM, 100 nM, 1 μM) for different time periods (10, 30, 60, 120, 240, and 480 min). Time-course experiments were performed in six replicates. Cells were harvested with PBS buffer, washed, and centrifuged at 1,500 x g for 5 min. A 200 μl aliquot of trizol reagent (9.5 g guanidinium thiocyanate, 3.1 g ammonium thiocyanate, 3.5 ml of 3 M sodium acetate, 5 g glycerol, 48 ml Roti Aqua-Phenol in 100 ml total volume) was added to the cell pellets to facilitate cell lysis and at the same time inactivate cAMP-degrading enzymes. Non-lysed cells and cell debris were removed by a centrifugation step using 30-kDa molecular weight cut-off filters (Amicon Millipore), which was performed at 14,000.0 x g for 10 minutes. Filtrates were stored at 4°C until they were analyzed by LC-MS/MS.
Publication 2023
ammonium thiocyanate Buffers Caffeine Cells Centrifugation Cloning Vectors Culture Media Eagle Enzymes Fetal Bovine Serum Glycerin guanidine thiocyanate istradefylline Pellets, Drug Phenol Sodium Acetate Tandem Mass Spectrometry Theophylline trizol
The cocrystals HKA·imidazole,
HKA·4-pyridone, and the complex HKA·4-aminopyridine were
obtained by ball milling equimolar quantities of HKA (0.5 mmol) and
coformer (0.5 mmol), in a 5 mL agate jar in the presence of two drops
(100 μL) of water, and two 3 mm agate balls, for 60 min in a
Retsch MM200 ball miller, operated at a frequency of 20 Hz. [H2PIP][KA]2·2H2O was obtained in
the same ball milling conditions as HKA·imidazole and HKA·4-pyridone,
but two drops of ethanol had to be added to the reacting mixture.
The complex (HKA)4·(theophylline)3 can
be produced by ball milling in the presence of two drops of methanol,
ethanol, acetone, ethyl acetate, or acetonitrile. HKA·DABCO was
obtained by manual grinding for 10 min, with an agate mortar and pestle,
and equimolar amounts of HKA (0.5 mmol) and DABCO (0.5 mmol), either
in the absence of solvent or in the presence of two drops of water,
ethanol, or methanol.
Publication 2023
Acetone acetonitrile Aminopyridines Ethanol ethyl acetate imidazole Methanol Pyridones Solvents Theophylline triethylenediamine
TGA measurements for
the crystalline complexes (HKA)4·(theophylline)3 and HKA·4-aminopyridine were performed with a PerkinElmer
TGA7 in the temperature range 40–300/500 °C under N2 gas flow at a heating rate of 5.00 °C min–1.
Publication 2023
Aminopyridines Theophylline
The crystalline
complex of HKA with
theophylline was obtained by slurry a 4:3 stoichiometric ratio mixture
of HKA (1.2 mmol) and theophylline (0.9 mmol) in 5 mL of methanol,
ethanol, acetone, ethyl acetate, or acetonitrile. (HKA)2·urotropine was obtained by slurry a 2:1 stoichiometric ratio
mixture of HKA (1.2 mmol) and urotropine (0.6 mmol) in a solution
of 5 mL of ethanol and 1 mL of water.
Publication 2023
Acetone acetonitrile Ethanol ethyl acetate Methanol Theophylline

Top products related to «Theophylline»

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Theophylline is a laboratory equipment product manufactured by Merck Group. It is a crystalline compound commonly used as a standard or reference material in analytical procedures. Theophylline is a key component in various analytical techniques, such as chromatography and spectroscopy, where it serves as a reference point for identification and quantification of similar compounds.
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Caffeine is a naturally occurring stimulant compound that can be extracted and purified for use in various laboratory applications. It functions as a central nervous system stimulant, inhibiting the action of adenosine receptors in the brain.
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Theobromine is a chemical compound that is commonly used in laboratory settings. It is a naturally occurring alkaloid found in cocoa beans, tea leaves, and other plants. Theobromine is a stimulant and has been studied for its potential therapeutic benefits, but its core function is as a laboratory reagent and analytical tool.
Sourced in United States, Belgium
Paraxanthine is a chemical compound used in various laboratory applications. It is a methylxanthine derivative with a structural similarity to caffeine. Paraxanthine is primarily utilized as a research tool and standard in analytical chemistry, pharmaceutical development, and biochemical studies.
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Fetal Bovine Serum (FBS) is a cell culture supplement derived from the blood of bovine fetuses. FBS provides a source of proteins, growth factors, and other components that support the growth and maintenance of various cell types in in vitro cell culture applications.
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Acetonitrile is a colorless, volatile, flammable liquid. It is a commonly used solvent in various analytical and chemical applications, including liquid chromatography, gas chromatography, and other laboratory procedures. Acetonitrile is known for its high polarity and ability to dissolve a wide range of organic compounds.
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Catechin is a natural polyphenolic compound found in various plants, including green tea. It functions as an antioxidant, with the ability to scavenge free radicals and protect cells from oxidative stress.
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Sodium hydroxide is a chemical compound with the formula NaOH. It is a white, odorless, crystalline solid that is highly soluble in water and is a strong base. It is commonly used in various laboratory applications as a reagent.
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DMSO is a versatile organic solvent commonly used in laboratory settings. It has a high boiling point, low viscosity, and the ability to dissolve a wide range of polar and non-polar compounds. DMSO's core function is as a solvent, allowing for the effective dissolution and handling of various chemical substances during research and experimentation.
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Epicatechin is a natural compound found in various plants and is commonly used in laboratory settings. It serves as a standard reference material for analytical and research purposes. Epicatechin exhibits antioxidant properties and is often employed in the evaluation of antioxidant activity and the development of analytical methods.

More about "Theophylline"

Theophylline is a versatile methylxanthine compound with a range of pharmacological properties, including bronchodilation, respiratory stimulation, and potential effects on the central nervous and cardiovascular systems.
It is commonly used in the management of asthma, chronic obstructive pulmonary disease (COPD), and other respiratory conditions.
Theophylline works by relaxing the airway muscles, improving airflow, and reducing inflammation.
Researchers can optimize their theophylline studies using PubCompare.ai, an innovative AI-driven platform that compares research protocols to identify the most reproducible and accurate procedures from the literature, preprints, and patents.
This intuitive tool streamlines protocol selection and comparison, enhancing the efficiency and quality of theophylline research.
Theophylline is structurally related to other methylxanthines, such as caffeine and theobromine, which share similar pharmacological properties.
Paraxanthine is a primary metabolite of theophylline and can also exhibit bronchodilatory effects.
Proper dosing and monitoring are crucial for theophylline, as it has a narrow therapeutic window and can interact with other medications.
In theophylline research, common experimental techniques may involve the use of fetal bovine serum (FBS), acetonitrile, catechin, sodium hydroxide, dimethyl sulfoxide (DMSO), and epicatechin, among others.
Researchers should carefully consider the selection and optimization of these reagents and methodologies to ensure the reproducibility and accuracy of their findings.