A quick inexpensive laboratory method in acute paracetamol poisoning could improve risk assessment, management and resource utilization

Objectives:

Acute paracetamol poisoning is an emerging problem in Sri Lanka. Management guidelines recommend ingested dose and serum paracetamol concentrations to assess the risk. Our aim was to determine the usefulness of the patient''s history of an ingested dose of >150 mg/kg and paracetamol concentration obtained by a simple colorimetric method to assess risk in patients with acute paracetamol poisoning.

Materials and Methods:

Serum paracetamol concentrations were determined in 100 patients with a history of paracetamol overdose using High Performance Liquid Chromatography (HPLC); (reference method). The results were compared to those obtained with a colorimetric method. The utility of risk assessment by reported dose ingested and colorimetric analysis were compared.

Results:

The area under the receiver operating characteristic curve for the history of ingested dose was 0.578 and there was no dose cut-off providing useful risk categorization. Both analytical methods had less than 5% intra- and inter-batch variation and were accurate on spiked samples. The time from blood collection to result was six times faster and ten times cheaper for colorimetry (30 minutes, US$2) than for HPLC (180 minutes, US$20). The correlation coefficient between the paracetamol levels by the two methods was 0.85. The agreement on clinical risk categorization on the standard nomogram was also good (Kappa = 0.62, sensitivity 81%, specificity 89%).

Conclusions:

History of dose ingested alone greatly over-estimated the number of patients who need antidotes and it was a poor predictor of risk. Paracetamol concentrations by colorimetry are rapid and inexpensive. The use of these would greatly improve the assessment of risk and greatly reduce unnecessary expenditure on antidotes.KEY WORDS: Acute poisoning, paracetamol concentration, risk assessment     

Comparison of analgesic effects of nimesulide, paracetamol, and their combination in animal models

Objectives:

To compare the analgesic activity of nimesulide and paracetamol alone and their combination in animal models for the degree of analgesia and the time course of action.

Materials and Methods:

Analgesia was studied in albino rats using formalin test and in albino mice using writhing test and the radiant heat method. For each test, four groups of six animals each were orally fed with a single dose of nimesulide, paracetamol, and combination of nimesulide + paracetamol and gum acacia as control, respectively.

Results:

In all the three test models, all three drug treatments showed significant analgesia (P < 0.001) as compared to control, but there was no significant difference in the analgesia produced by either drugs alone or in combination. The radiant heat method demonstrated a quicker onset and longer duration of action with nimesulide, whereas writhing test showed a quicker onset of action with paracetamol. In formalin test, greater degree of analgesia was seen with individual drugs than that of the combination, though this difference was not statistically significant.

Conclusions:

Nimesulide and paracetamol combination offers no advantage over nimesulide alone or paracetamol alone, either in terms of degree of analgesia or onset of action. Therefore, our study supports the reports claiming irrationality of the fixed dose combination of nimesulide and paracetamol.     

The effect of intracerebroventricular injection of histamine in visceral nociception induced by acetic acid in rats

Objective:

This study was designed to investigate the role of brain histamine and H1 and H2 receptors in mediating the central perception of visceral pain in rats.

Materials and Methods:

In conscious rats implanted with a lateral brain ventricle cannula, the effect of intracerebroventricular (i.c.v.) injection of histamine (2.5, 10, and 40 μg), and chlorpheniramine and ranitidine at the same doses of 5, 20, and 80 μg were investigated on visceral pain. Visceral nociception induced by intraperitoneal (i.p.) injection of acetic acid (1 mL, 1%), and the number of complete abdominal wall muscle contractions accompanied with stretching of hind limbs (writhes) were counted for 1 h.

Results:

Histamine at doses of 10 and 40 μg and chlorpheniramine and ranitidine at the same doses of 20 and 80 μg, significantly decreased the numbers of writhes (P < 0.05). Pretreatment with chlorpheniramine and ranitidine at the same dose of 80 μg, significantly prevented histamine (40 μg)-induced antinociception (P < 0.05).

Conclusion:

The results of this study suggest that brain histamine may be involved in modulation of visceral antinociception through both central H₁and H₂receptors.     

Effect of acute paracetamol overdose on changes in serum and urine electrolytes

What is already known about this subject

• Paracetamol causes renal failure in overdose. Experimental studies have shown that paracetamol can inhibit COX II systemically in a manner similar to selective COX-II inhibitors.
• In overdose nonsteroidal anti-inflammatory drugs such as ibuprofen, cause dose-dependent increase in urinary potassium excretion (FeK) and sodium retention, probably due to vasoconstriction.

What this study adds

• Paracetamol overdose is associated with dose-related hypokalaemia and kaliuresis of short duration (<24 h), suggesting a specific renal effect of paracetamol in overdose.
• This effect seems likely to be via cyclo-oxygenase inhibition and may be separate from the nephrotoxic effects of paracetamol.

Aims

To investigate the effects of acute paracetamol overdose on renal function, serum and urine electrolyte excretion in man.

Methods

Two studies were performed in patients admitted with paracetamol overdose: a retrospective study examining changes in serum electrolytes, and a prospective study evaluating changes in serum and urine electrolytes. A control group with SSRI overdose was included in the prospective study.

Results

There was a significant dose-dependent relationship between admission (4 h) paracetamol concentration and fall in serum potassium in the retrospective study (P < 0.01) and a significant positive relationship between serum paracetamol at 4 h and fractional excretion of potassium at 12 h postingestion (P < 0.01) in the prospective study. No changes were seen in the control group. No cases developed renal failure.

Conclusions

Paracetamol overdose is associated with dose-related hypokalaemia, and kaliuresis of short duration (<24 h), suggesting a specific renal effect of paracetamol in overdose perhaps via cyclo-oxygenase inhibition. This effect seems distinct from any nephrotoxic effect of paracetamol.     

Effect of adrenergic blockers,carvedilol, prazosin,metoprolol and combination of prazosin and metoprolol on paracetamol-induced hepatotoxicity in rabbits

Objectives:

To evaluate hepatoprotective potential of carvedilol, prazosin, metoprolol and prazosin plus metoprolol in paracetamol-induced hepatotoxicity.

Materials and Methods:

Thirty-six male rabbits were divided into six groups, six in each, group 1 received distilled water, group 2 were treated with paracetamol (1 g/kg/day, orally), group 3, 4,5 and 6 were treated at a dose in (mg/kg/day) of the following: Carvedilol (10 mg), prazosin (0.5 mg), metoprolol (10 mg), and a combination of metoprolol (10 mg) and prazosin (0.5 mg) respectively 1 h before paracetamol treatment. All treatments were given for 9 days; animals were sacrificed at day 10. Liver function tests, malondialdehyde (MDA) and glutathione (GSH) in serum and liver homogenates were estimated. Histopathological examinations of liver were performed.

Results:

Histopathological changes of hepatotoxicity were found in all paracetamol-treated rabbits. The histopathological findings of paracetamol toxicity disappeared in five rabbits on prazosin, very mild in one. In carvedilol group paracetamol toxicity completely disappeared in three, while mild in three rabbits. Paracetamol hepatotoxicity was not changed by metoprolol. In metoprolol plus prazosin treated rabbits, moderate histopathological changes were observed. Serum liver function tests and MDA in serum and in liver homogenate were elevated; GSH was depleted after paracetamol treatment and returned back to the control value on prior treatment with prazosin. MDA in serum and liver homogenate, alkaline phosphatase, total bilirubin were significantly decreased after carvedilol and prazosin plus metoprolol treatments.

Conclusion:

Carvedilol and prazosin are hepatoprotective in paracetamol hepatotoxicity, combination of prazosin and metoprolol have moderate, and metoprolol has a little hepatoprotection.KEY WORDS: Antioxidant, carvedilol, liver toxicity, metoprolol, paracetamol, prazosin     

Inhibition of paracetamol glucuronidation by tyrosine kinase inhibitors

AIMS

We aimed to investigate the effects of tyrosine kinase inhibitors (TKIs) on paracetamol (acetaminophen) glucuronidation.

METHODS

The inhibition of nine small molecule TKIs on paracetamol glucuronidation was investigated in human liver microsomes (HLMs) and recombinant human UDP-glucuronosyltransferases (UGTs).

RESULTS

Sorafenib, dasatinib and imatinib exhibited mixed inhibition against paracetamol glucuronidation in pooled HLMs, and potent inhibition in UGT1A9 and UGT2B15. Dasatinib and imatinib also inhibited UGT1A1-mediated paracetamol glucuronidation. Axitinib, erlotinib, gefitinib, lapatinib, nilotinib and vandetanib exhibited weak inhibition of paracetamol glucuronidation activity in HLMs.

CONCLUSIONS

The inhibition of paracetamol glucuronidation by TKIs might be of particular concern when they are co-administered.     

Anti-hyperbilirubinemic and wound healing activity of aqueous extract of Calotropis procera leaves in Wistar rats

Aims:

The aim of this study was to evaluate the bilirubin lowering and wound healing property of aqueous extract of Calotropis procera (AECP) leaves in Wistar rats.

Materials and Methods:

Albino Wistar rats of either sex were used for the study. Bilirubin lowering property of C. procera leaves was evaluated using phenylhydrazine and paracetamol as inducing agents followed by measuring the concentration of serum total bilirubin in hyperbilirubinemic rats. Wound healing property was evaluated using incision and excision models by measuring tensile breaking strength, percentage wound contractions, and epithelization days, respectively.

Statistical Analysis:

Statistical comparison between groups in each experiment was done with one-way analysis of variance followed by Dunnett''s test.

Results:

AECP showed a significant (P < 0.05) decrease in concentrations of serum total bilirubin in hyperbilirubinemic rats as well as significant (P < 0.05) increase in breaking strength and percentage wound contractions with decreased epithelization period when compared to control groups.

Conclusions:

AECP showed significant bilirubin lowering and wound healing property in Wistar rats.KEY WORDS: Calotropis procera, excision, hyperbilirubinemia, incision, paracetamol, phenylhydrazine, wound healing     

An evaluation of vardenafil as a calcium channel blocker in pulmonary artery in rats

Objective:

Vardenafil was reported to relax rat pulmonary artery through endothelium-dependent mechanisms. The aim of this in vitro study was to investigate other related mechanisms for this effect.

Materials and Methods:

Endothelium-intact and denuded artery rings were suspended in order to record isometric tension. In the rings with or without endothelium, the concentration-response curves for vardenafil were generated. In the rings without endothelium the contractile response induced by phenylephrine (Phe) or KCl was assessed in the presence or absence of vardenafil. In the last set of experiments, pulmonary artery rings were exposed to calcium-free isotonic depolarizing solution and the contractile response induced by the addition of calcium was evaluated in the presence or absence of vardenafil, nifedipine, verapamil or 1H-[1,2,4] oxadiazolo[4,3-a] quinoxalin-1-one (ODQ).

Results:

Vardenafil attenuated pulmonary artery contraction induced by phenylephrine in the presence and absence of endothelium. In addition, vardenafil attenuated both Phe or KCl-induced contraction but, it''s effect on the KCl dose-response curve was more significant. Vardenafil also inhibited the contractile response induced by calcium in a dose-dependent manner. Addition of nifedipine or verapamil did not significantly alter this effect while ODQ incubation significantly inhibited vardenafil-induced relaxation.

Conclusion:

From these findings, it was proposed that vardenafil relaxed rat pulmonary artery through inhibiting calcium influx.KEY WORDS: Calcium, phosphodiesterase type 5 inhibitors, pulmonary artery, vardenafil     

Effects of imatinib mesylate on the pharmacokinetics of paracetamol (acetaminophen) in Korean patients with chronic myelogenous leukaemia

AIMS

The major objective of the present study was to investigate the effect of imatinib on the pharmacokinetics of paracetamol in patients with chronic myelogenous leukaemia (CML).

METHODS

Patients (n= 12) received a single oral dose of acetaminophen 1000 mg on day 1 (control). On days 2–8, imatinib 400 mg was administered daily. On day 8 (treatment), another 1000 mg dose of paracetamol was administered 1 h after the morning dose of imatinib 400 mg. Blood and urine samples were collected for bioanalytical analyses.

RESULTS

The area under the plasma concentration–time curve (AUC) for paracetamol, paracetamol glucuronide and paracetamol sulphate under control conditions was similar to that after treatment with imatinib; the 90% confidence interval of the log AUC ratio was within 0.8 to 1.25. Urinary excretion of paracetamol, paracetamol glucuronide and paracetamol sulphate was also unaffected by imatinib. The pharmacokinetics of paracetamol and imatinib in Korean patients with CML were similar to previous pharmacokinetic results in white patients with CML. Co-administration of a single dose of paracetamol and multiple doses of imatinib was well tolerated and safety profiles were similar to those of either drug alone.

CONCLUSIONS

The pharmacokinetics of paracetamol and its major metabolites in the presence of imatinib were similar to those of the control conditions and the combination was well tolerated. These findings suggest that imatinib can be safely administered with paracetamol without dose adjustment of either drug.     

Effect of phenolic acids on functions of rat aorta, vas deferens and on metabolic changes in streptozotocin-induced diabetes

Objectives:

This study aimed to investigate the effects of antioxidant treatment on streptozotocin (STZ)-induced diabetic metabolic and smooth muscle (SM) complications in rats.

Materials and Methods:

Threeweeks after STZ injection (i.v.), vehicle, p-OH benzoic (p-OHBA), protocatechic (PA) and gallic acids (GA) were separately administered (10 mg/kg each, i.p.) to the rats everyday for 3 weeks. Metabolic functions were observedregularly. The rats in all groups were sacrificed andaorta and Vas deferens were dissected. Theresponses of isolated organs to agonists (acetylcholine and phenylephrine) were recorded.

Results:

Protocatechic acid prevented increase in food consumption and feces output significantly. The responses of isolated organs to agonists increased in the STZ-diabetic rats. The test drugs either prevented, exacerbated or didnot affect the SMchanges in the STZ-diabetic rats.

Conclusions:

It was concluded that p-OHBA, PA and GA may cause effects independently of their antioxidant effect and/or of diabeticcomplications. They may exhibit pro-oxidant activities in the experimental conditions applied.KEY WORDS: Aorta, diabetic complications, oxidative stress, phenolic acids, Vas deferens     

Effect of acetaminophen (paracetamol) on human osteosarcoma cell line MG63

Aim:

To examine the effects of acetaminophen (paracetamol), a nonsteroidal anti-inflammatory drug (NSAID), on different cellular and functional parameters of the human osteosarcoma cell line MG63.

Methods:

Flow cytometry was used to study proliferation, antigenic profile, and phagocytic activity, and radioimmunoassay was used to determine osteocalcin synthesis as a cell differentiation marker.

Results:

Short-term treatment with therapeutic doses of paracetamol(5 or 25 μmol/L) reduced cell proliferation, osteocalcin synthesis, and phagocyte activity, and increased the expression of antigens involved in antigen presentation to T lymphocytes (CD80, CD86, HLA-DR).

Conclusion:

These findings suggest that paracetamol activates the osteoblast, inducing its immunogenic action to the detriment of its bone formation capacity.     

Visualization and Non-Destructive Quantification of Inkjet-Printed Pharmaceuticals on Different Substrates Using Raman Spectroscopy and Raman Chemical Imaging

Purpose

The purpose of this study was to investigate the applicability of Raman spectroscopy for visualization and quantification of inkjet-printed pharmaceuticals.

Methods

Haloperidol was used as a model active pharmaceutical ingredient (API), and a printable ink base containing lactic acid and ethanol was developed. Inkjet printing technology was used to apply haloperidol ink onto three different substrates. Custom-made inorganic compacts and dry foam, as well as marketed paracetamol tablets were used as the substrates.

Results

Therapeutic personalized doses were printed by using one to ten printing rounds on the substrates. The haloperidol content in the finished dosage forms were determined by high-performance liquid chromatography (HPLC). The distribution of the haloperidol on the dosage forms were visualized using Raman chemical imaging combined with principal components analysis (PCA). Raman spectroscopy combined with modeling by partial least squares (PLS) regression was used for establishment of a quantitative model of the haloperidol content in the printed dosage forms. A good prediction of the haloperidol content was achieved for the inorganic compacts, while a slightly poorer prediction was observed for the paracetamol tablets. It was not possible to quantify haloperidol on the dry foam due to the low and varying density of the substrate.

Conclusions

Raman spectroscopy is a useful tool for visualization and quality control of inkjet printed personalized medicine.

     

Antipyretic and antibacterial activity of Chloranthus erectus (Buch.-Ham.) Verdcourt leaf extract: A popular folk medicine of Arunachal Pradesh

Objective:

The main objective of this work was to study the antipyretic and antibacterial activity of C. erectus (Buch.-Ham.) Verdcourt leaf extract in an experimental albino rat model.

Materials and Methods:

The methanol extract of C. erectus leaf (MECEL) was evaluated for its antipyretic potential on normal body temperature and Brewer’s yeast-induced pyrexia in albino rat’s model. While the antibacterial activity of MECEL against five Gram (−) and three Gram (+) bacterial strains and antimycotic activity was investigated against four fungi using agar disk diffusion and microdilution methods.

Result

Yeast suspension (10 mL/kg b.w.) elevated rectal temperature after 19 h of subcutaneous injection. Oral administration of MECEL at 100 and 200 mg/kg b.w. showed significant reduction of normal rectal body temperature and yeast-provoked elevated temperature (38.8 ± 0.2 and 37.6 ± 0.4, respectively, at 2–3 h) in a dose-dependent manner, and the effect was comparable to that of the standard antipyretic drug–paracetamol (150 mg/kg b.w.). MECEL at 2 mg/disk showed broad spectrum of growth inhibition activity against both groups of bacteria. However, MECEL was not effective against the yeast strains tested in this study.

Conclusion

This study revealed that the methanol extract of C. erectus exhibited significant antipyretic activity in the tested models and antibacterial activity as well, and may provide the scientific rationale for its popular use as antipyretic agent in Khamptis’s folk medicines.     

Student evaluation of teaching and assessment methods in pharmacology

Background:

The students are in the best position to comment on the effectiveness of any teaching system and they may be regarded as the best judges to assess the teaching and evaluation methods.

Objective:

This study was designed to obtain student feedback on teaching and assessment methods in the subject of pharmacology and use it for improvement.

Materials and Methods:

Based on student feedback from batch 2006, innovative strategies were implemented. To know the effect of these strategies feedback was obtained from subsequent batch 2007 using a written validated questionnaire covering various aspects of teaching and assessment methods.

Results:

Students were satisfied with all teaching methods except lecture, seminars and pharmacy exercises. Majority of the students showed preference for tutorials, short answer questions and revision classes. All students felt that there should be more time for clinical pharmacology and bedside teaching. The performance score of the students (batch 2007) indicated improvement in their scores (12%) when earlier feedback suggestions were implemented. The pass percentage of the subsequent batch in university examinations improved from 90 to 100%.

Conclusion:

The implementation of suggestions obtained from students resulted in improvement in their performance. Hence, it is very essential to synchronize teaching and evaluation methods with special requirements of medical students.     

A systematic review of the effect of paracetamol on blood pressure in hypertensive and non-hypertensive subjects

Aim

To review current evidence on the effect of paracetamol on blood pressure (BP), the quality of the previous studies and the validity of the results, and to summarize these findings.

Methods

A systematic literature review was performed by searching PubMed, the Cochrane library and EMBASE for publications between the years 1963 and 2012.

Results

We identified three case reports, seven prospective observational trials, six randomized controlled trials, one commentary and two reviews. Some, but not all, of the observational studies, which included over 147 000 patients, showed an increased risk of hypertension with paracetamol use. The randomized studies were generally small and the results were inconsistent. Three studies, which included 104 patients, showed an increase of systolic BP by ∼4 mmHg, two studies, which included 27 patients, reported no change in BP and one study, which included 21 patients, reported a fall in BP although no placebo arm was included for comparison.

Conclusions

The overall effect of paracetamol on BP is unclear. Given that paracetamol is often suggested as a safer alternative to non-steroidal anti-inflammatory drugs (NSAIDs), it would seem that further prospective evidence is now needed to address the effect of paracetamol on BP. This would be best done with larger studies in relevant cohorts using BP measured by ambulatory BP monitoring as the primary endpoint.     

Disposition kinetics of sparfloxacin in healthy, hepatopathic, and nephropathic conditions in chicken after single intravenous administration

Objective:

To study the variation of disposition kinetic values of sparfloxacin in healthy, hepatopathic, and nephropathic chickens after a single intravenous administration.

Materials and Methods:

Hepatotoxicity was induced by the administration of paracetamol (500 mg / kg / day, p.o. for seven days) and nephrotoxicity by uranyl nitrate (2.0 mg / kg / day dissolved in distilled water, i.v. for four days) in chickens. Disposition kinetic studies of sparfloxacin were investigated in healthy as well as hepatopathic and nephropathic birds after a single intravenous administration at 40 mg / kg body weight.

Results:

Maximum plasma concentration detected at 0.16 hour was 31.25 ± 2.95, 61.95 ±1.85, and 99.86 ± 2.21 μg / ml in healthy, hepatopathic, and nephropathic group, respectively. The drug could not be detected in the plasma of healthy birds beyond 12-hour period, while the same was detectable for 72 hour in the plasma of hepatopathic and nephropathic birds. The concentration of sparfloxacin was significantly (P < 0.01) higher in all the samples of hepathopathic and nephropathic birds compared to healthy birds. All the kinetic values were increased (P < 0.01) in the hepatopathic and nephropathic birds, except Vd_area and Cl_B values in hepatopathic Birds; while β and Cl_B values nephropathic birds were decreased significantly than that of healthy birds.

Conclusions:

The dose of sparfloxacin may be reduced in hepatopathic as well as nephropathic birds.     

Development of an in vitro cell culture model to study milk to plasma ratios of therapeutic drugs

Objective:

To create an in vitro cell culture model to predict the M/P (concentration of drug in milk/concentration in maternal plasma) ratios of therapeutic drugs viz. rifampicin, theophylline, paracetamol, and aspirin.

Materials and Methods:

An in vitro cell culture model using CIT3 cells (mouse mammary epithelial cells) was created by culturing the cells on transwells. The cells formed an integral monolayer, allowing only transcellular transport as it happens in vivo. Functionality of the cells was confirmed through scanning electron microscopy. Time wise transfer of the study drugs from plasma to milk was studied and compared with actual (in vivo) M/P ratios obtained at reported t_max for the respective drugs.

Results:

The developed model mimicked two important intrinsic factors of mammary epithelial cells viz. secretory and tight-junction properties and also the passive route of drug transport. The in vitro M/P ratios at reported t_max were 0.23, 0.61, 0.87, and 0.03 respectively, for rifampicin, theophylline, paracetamol, and salicylic acid as compared to 0.29, 0.65, 0.65, and 0.22, respectively, in vitro.

Conclusion:

Our preliminary effort to develop an in vitro physiological model showed promising results. Transfer rate of the drugs using the developed model compared well with the transfer potential seen in vivo except for salicylic acid, which was transferred in far lower concentration in vitro. The model has a potential to be developed as a non-invasive alternative to the in vitro technique for determining the transfer of therapeutic drugs into breast milk.KEY WORDS: Cell culture, milk-plasma ratio, CIT3 cells, in vitro, M/P ratios, reversed phase-High performance liquid chromatography     

An efficient procedure for purification of recombinant human β heat shock protein 90

Background and the purpose of the study

Heat Shock Protein 90 (Hsp90) is typically the most abundant chaperone in the eukaryotic cell cytoplasm, and its expression is essential for loading immunogenic peptides onto major histocompatibility complex molecules for presentation to T-cells. Therefore, it may act as a good candidate as an adjuvant molecule in vaccine technology.

Methods

Initially the human Hsp90β gene was cloned into the heat inducible expression vector pGP1-2 and then the recombinant protein was isolated by ion exchange chromatography. After intradermal injection of confirmed purified band of protein to rabbits and isolation of the serum IgG antibody, for its affinity purification, the rabbit''s purified Hsp90 specific IgG was coupled to the cyanogen bromide-activated Sepharose 4B.

Results

The recovery of the purified protein of interest by affinity chromatography was 50%.

Conclusion

This research enabled purification of human heat shock protein by a laboratory prepared column chromatography.