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Molecular distribution, sources and potential health risks of fine particulate-bound polycyclic aromatic hydrocarbons during high pollution episodes in a subtropical urban city

Singh A, Banerjee T, Latif MT, Ramanathan S, Suradi H, Othman M, et al.

Chemosphere, 2023 Nov;340:139943.
PMID: 37625487 DOI: 10.1016/j.chemosphere.2023.139943

Abundance of fine particulate-bound 16 priority polycyclic aromatic hydrocarbons (PAHs) was investigated to ascertain its sources and potential carcinogenic health risks in Varanasi, India. The city represents a typical urban settlement of South Asia having particulate exposure manyfold higher than standard with reports of pollution induced mortalities and morbidities. Fine particulates (PM2.5) were monitored from October 2019 to May 2020, with 32% of monitoring days accounting ≥100 μgm-3 of PM2.5 concentration, frequently from November to January (99% of monitoring days). The concentration of 16 priority PAHs varied from 24.1 to 44.6 ngm-3 (mean: 33.1 ± 3.2 ngm-3) without much seasonal deviations. Both low (LMW, 56%) and high molecular weight (HMW, 44%) PAHs were abundant, with Fluoranthene (3.9 ± 0.4ngm-3) and Fluorene (3.5 ± 0.3ngm-3) emerged as most dominating PAHs. Concentration of Benzo(a)pyrene (B(a)P, 0.5 ± 0.1ngm-3) was lower than the national standard as it contributed 13% of total PAHs mass. Diagnostic ratios of PAH isomers indicate predominance of pyrogenic sources including emissions from biomass burning, and both from diesel and petrol-driven vehicles. Source apportionment using receptor model revealed similar observation of major PAHs contribution from biomass burning and fuel combustion (54% of source contribution) followed by coal combustion for residential heating and cooking purposes (44%). Potential toxicity of B[a]P equivalence ranged from 0.003 to 1.365 with cumulative toxicity of 2.13ngm-3. Among the PAH species, dibenzo[h]anthracene contributed maximum toxicity followed by B[a]P, together accounting 86% of PAH induced carcinogenicity. Incremental risk of developing cancer through lifetime exposure (ILCR) of PAHs was higher in children (3.3 × 10-4) with 56% contribution from LMW PAHs, primarily through ingestion and dermal contact. Adults in contrast, were more exposed to inhale airborne PAHs with cumulative ILCR of 2.2 × 10-4. However, ILCR to PM2.5 exposure is probably underestimated considering unaccounted metal abundance thus, require source-specific control measures.
Mitragynine (Kratom) impairs spatial learning and hippocampal synaptic transmission in rats

Hassan Z, Suhaimi FW, Ramanathan S, Ling KH, Effendy MA, Müller CP, et al.

J. Psychopharmacol. (Oxford), 2019 07;33(7):908-918.
PMID: 31081443 DOI: 10.1177/0269881119844186

BACKGROUND: Mitragynine is the major alkaloid of Mitragyna speciosa (Korth.) or Kratom, a psychoactive plant widely abused in Southeast Asia. While addictive effects of the substance are emerging, adverse cognitive effects of this drug and neuropharmacological actions are insufficiently understood.
AIMS: In the present study, we investigated the effects of mitragynine on spatial learning and synaptic transmission in the CA1 region of the hippocampus.
METHODS: Male Sprague Dawley rats received daily (for 12 days) training sessions in the Morris water maze, with each session followed by treatment either with mitragynine (1, 5, or 10 mg/kg; intraperitoneally), morphine (5 mg/kg; intraperitoneally) or a vehicle. In the second experiment, we recorded field excitatory postsynaptic potentials in the hippocampal CA1 area in anesthetized rats and assessed the effects of mitragynine on baseline synaptic transmission, paired-pulse facilitation, and long-term potentiation. Gene expression of major memory- and addiction-related genes was investigated and the effects of mitragynine on Ca2+ influx was also examined in cultured primary neurons from E16-E18 rats.
RESULTS/OUTCOMES: Escape latency results indicate that animals treated with mitragynine displayed a slower rate of acquisition as compared to their control counterparts. Further, mitragynine treatment significantly reduced the amplitude of baseline (i.e. non-potentiated) field excitatory postsynaptic potentials and resulted in a minor suppression of long-term potentiation in CA1. Bdnf and αCaMKII mRNA expressions in the brain were not affected and Ca2+ influx elicited by glutamate application was inhibited in neurons pre-treated with mitragynine.
CONCLUSIONS/INTERPRETATION: These data suggest that high doses of mitragynine (5 and 10 mg/kg) cause memory deficits, possibly via inhibition of Ca2+ influx and disruption of hippocampal synaptic transmission and long-term potentiation induction.
Method validation in quantitative analysis of phase I and phase II metabolites of mitragynine in human urine using liquid chromatography-tandem mass spectrometry

Lee MJ, Ramanathan S, Mansor SM, Yeong KY, Tan SC

Anal Biochem, 2018 02 15;543:146-161.
PMID: 29248503 DOI: 10.1016/j.ab.2017.12.021

A method using solid phase extraction and liquid chromatography-tandem mass spectrometry to quantitatively detect mitragynine, 16-carboxy mitragynine, and 9-O-demethyl mitragynine in human urine samples was developed and validated. The relevant metabolites were identified using multiple reaction monitoring in positive ionization mode using nalorphine as an internal standard. The method was validated for accuracy, precision, recovery, linearity, and lower limit of quantitation. The intra- and inter-day accuracy and precision were found in the range of 83.6-117.5% with coefficient of variation less than 13%. The percentage of recovery for mitragynine, 16-carboxy mitragynine, and 9-O-demethyl mitragynine was within the range of 80.1-118.9%. The lower limit of quantification was 1 ng/mL for mitragynine, 2 ng/mL for 16-carboxy mitragynine, and 50 ng/mL for 9-O-demethyl mitragynine. The developed method was reproducible, high precision and accuracy with good linearity and recovery for mitragynine, 16-carboxy mitragynine, and 9-O-demethyl mitragynine in human urine.
Kratom and Pain Tolerance: A Randomized, Placebo-Controlled, Double-Blind Study

Vicknasingam B, Chooi WT, Rahim AA, Ramachandram D, Singh D, Ramanathan S, et al.

Yale J Biol Med, 2020 06;93(2):229-238.
PMID: 32607084

Background: Kratom has a long history of traditional medicine use in Southeast Asia. Consumption of kratom products has also been reported in the US and other regions of the world. Pain relief is among many self-reported kratom effects but have not been evaluated in controlled human subject research. Methods: Kratom effects on pain tolerance were assessed in a randomized, placebo-controlled, double-blind study. During a 1-day inpatient stay, participants received a randomized sequence of kratom and placebo decoctions matched for taste and appearance. Pain tolerance was measured objectively in a cold pressor task (CPT) as time (seconds) between the pain onset and the hand withdrawal from the ice bath. Health status, vital signs, objective, and subjective indicators of withdrawal symptoms, self-reported data on lifetime kratom use patterns, and assessments of blinding procedures were also evaluated. Results: Twenty-six males with the mean (SD) age 24.3 (3.4) years were enrolled. They reported the mean (SD) 6.1 (3.2) years of daily kratom consumption. Pain tolerance increased significantly 1 hour after kratom ingestion from the mean (SD) 11.2 (6.7) seconds immediately before to 24.9 (39.4) seconds 1 hour after kratom consumption (F(2,53.7)=4.33, p=0.02). Pain tolerance was unchanged after consuming placebo drinks: 15.0 (19.0) seconds immediately before and 12.0 (8.1) seconds 1 hour after consumption of placebo (F(2,52.8)=0.93, p=0.40). No discomfort or signs of withdrawal were reported or observed during 10-20 hours of kratom discontinuation. Conclusions: Kratom decoction demonstrated a substantial and statistically significant increase in pain tolerance. Further rigorous research on kratom pain-relieving properties and a safety profile is needed.
Kratom (Mitragyna speciosa): worldwide issues

Ramanathan S, McCurdy CR

Curr Opin Psychiatry, 2020 07;33(4):312-318.
PMID: 32452943 DOI: 10.1097/YCO.0000000000000621

PURPOSE OF REVIEW: To inform readers about the increasingly popular Western dietary supplement, kratom (Mitragyna speciosa) and how the products are available in the Western world compared with traditional Southeast Asian use. Kratom has been traditionally used for increasing stamina of outdoor laborers (farmers), mood enhancement, pain, and opium addiction. Interestingly, kratom has been reported to have a paradoxical effect in that stimulant feelings, and sedative feelings can be obtained depending on the amount utilized. There are several biologically active alkaloids present in kratom.
RECENT FINDINGS: Recent studies have been focused on the interactions of mitragynine, the most abundant alkaloid, and opioid-like effects. This has been driven by the harm that kratom products have produced in the Western world, in stark contrast to the lack of harm in Southeast Asian traditional use over centuries. Many users in the Western world ingest kratom for mood enhancement and/or to ween themselves from prescription or illicit opioids. Highly concentrated products and recreational use and misuse have resulted in individuals pushing doses to levels that have not been imagined or ever studied in animal, let alone humans.
SUMMARY: Kratom, as a preparation and how it is utilized is different around the world.
Fulltext Investigation of the Adrenergic and Opioid Binding Affinities, Metabolic Stability, Plasma Protein Binding Properties, and Functional Effects of Selected Indole-Based Kratom Alkaloids

Obeng S, Kamble SH, Reeves ME, Restrepo LF, Patel A, Behnke M, et al.

J Med Chem, 2020 01 09;63(1):433-439.
PMID: 31834797 DOI: 10.1021/acs.jmedchem.9b01465

Selected indole-based kratom alkaloids were evaluated for their opioid and adrenergic receptor binding and functional effects, in vivo antinociceptive effects, plasma protein binding, and metabolic stability. Mitragynine, the major alkaloid in Mitragyna speciosa (kratom), had higher affinity at opioid receptors than at adrenergic receptors while the vice versa was observed for corynantheidine. The observed polypharmacology of kratom alkaloids may support its utilization to treat opioid use disorder and withdrawal.
Fulltext Investigation of antioxidant and hepatoprotective activity of standardized Curcuma xanthorrhiza rhizome in carbon tetrachloride-induced hepatic damaged rats

Devaraj S, Ismail S, Ramanathan S, Yam MF

ScientificWorldJournal, 2014;2014:353128.
PMID: 25133223 DOI: 10.1155/2014/353128

Curcuma xanthorrhiza (CX) has been used for centuries in traditional system of medicine to treat several diseases such as hepatitis, liver complaints, and diabetes. It has been consumed as food supplement and "jamu" as a remedy for hepatitis. Hence, CX was further explored for its potential as a functional food for liver related diseases. As such, initiative was taken to evaluate the antioxidant and hepatoprotective potential of CX rhizome. Antioxidant activity of the standardized CX fractions was determined using in vitro assays. Hepatoprotective assay was conducted against carbon tetrachloride- (CCl4-) induced hepatic damage in rats at doses of 125, 250, and 500 mg/kg of hexane fraction. Highest antioxidant activity was found in hexane fraction. In the case of hepatoprotective activity, CX hexane fraction showed significant improvement in terms of a biochemical liver function, antioxidative liver enzymes, and lipid peroxidation activity. Good recovery was observed in the treated hepatic tissues histologically. Hence, the results concluded that CX hexane fraction possessed prominent hepatoprotective activities which might be due to its in vitro antioxidant activity. These findings also support the use of CX as a functional food for hepatitis remedy in traditional medicinal system.
Intestinal permeability of mitragynine in rats using in situ absorption model

Jagabalan JDY, Murugaiyah V, Zainal H, Mansor SM, Ramanathan S

J Asian Nat Prod Res, 2019 Apr;21(4):351-363.
PMID: 29667422 DOI: 10.1080/10286020.2018.1461088

The intestinal permeability of mitragynine was investigated in situ using a single pass intestinal perfusion (SPIP) absorption model, in small intestine of rat using mitragynine in the absence/presence of the permeability markers, P-gp and/or CYP3A4 inhibitors. Mitragynine demonstrated high intestinal permeability (Peff of 1.11 × 10-4 cm/s) that is in the range of highly permeable drugs such as propranolol (Peff of 1.27 × 10-4 cm/s) indicating that it readily crosses the intestine. The addition of azithromycin (P-glycoprotein inhibitor) and ciprofloxacin (CYP3A4 inhibitor) or combination of both has no effect on intestinal permeability of mitragynine across the rat small intestine.
Fulltext Influence of sonication on the phenolic content and antioxidant activity of Terminalia catappa L. leaves

Annegowda HV, Anwar LN, Mordi MN, Ramanathan S, Mansor SM

Pharmacognosy Res, 2010 Nov;2(6):368-73.
PMID: 21713141 DOI: 10.4103/0974-8490.75457

This study was designed to evaluate the phenolic content and antioxidant activity of ethanolic extracts from T. catappa leaves obtained by different intervals of sonication.
Incomplete hypospadiac urethral duplication with posterior urethral valves

Ramanujam TM, Sergius A, Usha V, Ramanathan S

Pediatr Surg Int, 1998 Nov;14(1-2):134-7.
PMID: 9880724

Urethral duplication (UD) is an uncommon malformation. Obstruction rarely occurs in hypospadiac UD. We describe two children with incomplete hypospadiac UD in association with posterior urethral valves, a combination not previously recognised. The embryonic significance of this anomaly is discussed. Keywords Urethral duplication. Hypospadias. Posterior urethral valve. Megalourethra
Fulltext In vitro and in vivo effects of three different Mitragyna speciosa korth leaf extracts on phase II drug metabolizing enzymes--glutathione transferases (GSTs)

Azizi J, Ismail S, Mordi MN, Ramanathan S, Said MI, Mansor SM

Molecules, 2010 Jan 20;15(1):432-41.
PMID: 20110902 DOI: 10.3390/molecules15010432

In the present study, we investigate the effects of three different Mitragyna speciosa extracts, namely methanolic, aqueous and total alkaloid extracts, on glutathione transferase-specific activity in male Sprague Dawley rat liver cytosol in vitro and in vivo. In the in vitro study, the effect of Mitragyna speciosa extracts (0.01 to 750 microg/mL) against the specific activity of glutathione transferases was examined in rat liver cytosolic fraction from untreated rats. Our data show concentration dependent inhibition of cytosolic GSTs when Mitragyna speciosa extract was added into the reaction mixture. At the highest concentration used, the methanolic extract showed the highest GSTs specific activity inhibition (61%), followed by aqueous (50%) and total alkaloid extract (43%), respectively. In in vivo study, three different dosages; 50, 100 and 200 mg/kg for methanolic and aqueous extracts and 5, 10 and 20 mg/kg for total alkaloid extract were given orally for 14 days. An increase in GST specific activity was generally observed. However, only Mitragyna speciosa aqueous extract with a dosage of 100 mg/kg showed significant results: 129% compared to control.
Fulltext In vitro and in vivo anticandidal activity of Swietenia mahogani methanolic seed extract

Sahgal G, Ramanathan S, Sasidharan S, Mordi MN, Ismail S, Mansor SM

Trop Biomed, 2011 Apr;28(1):132-7.
PMID: 21602779 MyJurnal

Swietenia mahogani crude methanolic (SMCM) seed extract was investigated for the antifungal activity against Candida albicans which has not been evaluated previously. The antifungal activity was evaluated against C. albicans via disk diffusion, minimum inhibition concentration (MIC), scanning electron microscope (SEM), transmission electron microscope (TEM) and time killing profile. The MIC value of SMCM seed extract is 12.5 mg/ml. The SEM and TEM findings showed there is morphological changes and cytological destruction of C. albicans at the MIC value. Animal model was used to evaluate the in vivo antifungal activity of SMCM seed extract. The colony forming unit (CFU) were calculated per gram of kidney sample and per ml of blood sample respectively for control, curative and ketaconazole treated groups. There was significant reduction for the CFU/ml of blood and CFU/g of kidney. This indicated that the extract was observed to be effective against C. albicans in vitro and in vivo conditions.
Fulltext In Vitro antioxidant and xanthine oxidase inhibitory activities of methanolic Swietenia mahagoni seed extracts

Sahgal G, Ramanathan S, Sasidharan S, Mordi MN, Ismail S, Mansor SM

Molecules, 2009 Nov 06;14(11):4476-85.
PMID: 19924080 DOI: 10.3390/molecules14114476

This study examines the in vitro antioxidant activities of the methanol extract of Swietenia mahagoni seeds (SMCM seed extract). The extract was screened for possible antioxidant activities by free radical scavenging activity (DPPH), xanthine oxidase inhibition (XOI), hydrogen peroxide scavenging activity (HPSA) and ferric-reducing antioxidant power (FRAP) assays. The total phenolic and flavonoid contents were also determined. The extract exhibits antioxidant activity of 23.29% with an IC(50 )value of 2.3 mg/mL in the DPPH radical scavenging method, 47.2% in the XOI assay, 49.5% by the HPSA method, and 0.728 mmol/Fe(II)g in the FRAP method at the concentration tested. The amount of total phenolics and flavonoid contents was 70.83 mg gallic acid equivalent (GAE) and 2.5 +/- 0.15 mg of catechin equivalent per gram of dry extract, respectively. High Performance Thin Layer Chromatography (HPTLC) screening indicates the presence of phenolic compounds in the SMCM seed extract. The results indicate that the extract has both high free radical scavenging and xanthine oxidase inhibition activity. The antioxidant activity of SMCM seed extract is comparable with that of other Malaysian tropical fruits and herbal plants.
Fulltext Graphitic carbon nitride/graphene nanoflakes hybrid system for electrochemical sensing of DNA bases in meat samples

Kalaiyarasi J, Pandian K, Ramanathan S, Gopinath SCB

Sci Rep, 2020 07 30;10(1):12860.
PMID: 32732935 DOI: 10.1038/s41598-020-69578-8

This research presents a simple, fast and simultaneous electrochemical quantitative determination of nucleobases, for example guanine (G), adenine (A), and thymine (T) in a beef and chicken livers samples to measure the quality of food products based on hybrids of graphitic carbon nitride/Graphene nanoflakes (g-C3N4/GNF) modified electrode. Graphitic carbon nitride (g-C3N4) made of graphite-like covalent link connects nitrogen, nitride, and carbon atoms in the structural design with improved the electrical properties and low band gap semiconductor. The g-C3N4/GNF nanocomposite was synthesized by the hydrothermal treatment to form a porous g-C3N4 interconnected three dimensional (3D) network of g-C3N4 and GNF. The 3D g-C3N4/GNF/GCE was utilized for the detection of nucleic acid bases with a well resolved oxidation peak for the individual analyte. The electrocatalytic current was established to be a linear range from 0.3 × 10-7 to 6.6 × 10-6, 0.3 × 10-7 to 7.3 × 10-6, and 5.3 × 10-6 to 63.3 × 10-4 M for G, A, and T with a detection limit of 4.7, 3.5 and 55 nM, respectively. The diffusion co-efficient and the kinetic parameters were derived from the chronoamperometry technique. The proposed sensing strategy has been effectively used for the application in real sample analysis and observed that the electrode free from the surface fouling.
Genomic analyses in Cornelia de Lange Syndrome and related diagnoses: Novel candidate genes, genotype-phenotype correlations and common mechanisms

Kaur M, Blair J, Devkota B, Fortunato S, Clark D, Lawrence A, et al.

Am J Med Genet A, 2023 Aug;191(8):2113-2131.
PMID: 37377026 DOI: 10.1002/ajmg.a.63247

Cornelia de Lange Syndrome (CdLS) is a rare, dominantly inherited multisystem developmental disorder characterized by highly variable manifestations of growth and developmental delays, upper limb involvement, hypertrichosis, cardiac, gastrointestinal, craniofacial, and other systemic features. Pathogenic variants in genes encoding cohesin complex structural subunits and regulatory proteins (NIPBL, SMC1A, SMC3, HDAC8, and RAD21) are the major pathogenic contributors to CdLS. Heterozygous or hemizygous variants in the genes encoding these five proteins have been found to be contributory to CdLS, with variants in NIPBL accounting for the majority (>60%) of cases, and the only gene identified to date that results in the severe or classic form of CdLS when mutated. Pathogenic variants in cohesin genes other than NIPBL tend to result in a less severe phenotype. Causative variants in additional genes, such as ANKRD11, EP300, AFF4, TAF1, and BRD4, can cause a CdLS-like phenotype. The common role that these genes, and others, play as critical regulators of developmental transcriptional control has led to the conditions they cause being referred to as disorders of transcriptional regulation (or "DTRs"). Here, we report the results of a comprehensive molecular analysis in a cohort of 716 probands with typical and atypical CdLS in order to delineate the genetic contribution of causative variants in cohesin complex genes as well as novel candidate genes, genotype-phenotype correlations, and the utility of genome sequencing in understanding the mutational landscape in this population.
Formulation and evaluation of wound healing activity of Elaeis guineensis Jacq leaves in a Staphylococcus aureus infected Sprague Dawley rat model

Rajoo A, Ramanathan S, Mansor SM, Sasidharan S

J Ethnopharmacol, 2021 Feb 10;266:113414.
PMID: 32980488 DOI: 10.1016/j.jep.2020.113414

ETHNOPHARMACOLOGICAL RELEVANCE: Medicinal plants are crucial to healing numerous illnesses. Elaeis guineensis Jacq (family Arecaceae) is a medicinal plant traditionally used for the treatment of wounds.
AIM OF THE STUDY: However, there are no scientific reports documented on the wound healing activities of this plant against Staphylococcus aureus infections in the Sprague Dawley male rat model. Thus, the present study was conducted to evaluate the wound healing potential of E. guineensis extract leaves.
MATERIALS AND METHODS: The crude extract was prepared in 10% (w/w) ointment and evaluated for wound healing activity using excision and infected wound models in Sprague Dawley rats. The wound healing activity was evaluated from wound closure rate, CFU reduction, histological analysis of granulation tissue and matrix metalloprotease expression.
RESULTS: The results show that the E. guineensis extract has potent wound healing ability, as manifest from improved wound closure and tissue regeneration supported by histopathological parameters. Assessment of granulation tissue every fourth day showed a significant reduction in the microbial count. The expression of matrix metalloproteinases was well correlated with the other results, hence confirming E. guineensis wound healing activity's effectiveness.
CONCLUSIONS: E. guineensis enhanced infected wound healing in rats, thus supporting its traditional use.
Exploring the Chemistry of Alkaloids from Malaysian Mitragyna speciosa (Kratom) and the Role of Oxindoles on Human Opioid Receptors

Chear NJ, León F, Sharma A, Kanumuri SRR, Zwolinski G, Abboud KA, et al.

J Nat Prod, 2021 04 23;84(4):1034-1043.
PMID: 33635670 DOI: 10.1021/acs.jnatprod.0c01055

Ten indole and oxindole alkaloids (1-10) were isolated from the freshly collected leaves of Malaysian Mitragyna speciosa (Kratom). The chemical structures of these compounds were established on the basis of extensive 1D and 2D NMR and HRMS data analysis. The spectroscopic data of mitragynine oxindole B (4) are reported herein for the first time. The spatial configuration of mitragynine oxindole B (4) was confirmed by single-crystal X-ray diffraction. Simultaneous quantification of the isolated alkaloids in the M. speciosa leaf specimens collected from different locations in the northern region of Peninsular Malaysia was also performed using UPLC-MS/MS. The oxindole alkaloids (1-4) and the indole alkaloid (10) were assessed for binding affinity at opioid receptors. Corynoxine (1) showed high binding affinity to μ-opioid receptors with a Ki value of 16.4 nM. Further, corynoxine (1) was 1.8-fold more potent than morphine in rats subjected to a nociceptive hot plate assay. These findings have important implications for evaluating the combined effects of the minor oxindole alkaloids in the overall therapeutic activity of M. speciosa.
Evaluation of toxicity profile of kratom (Mitragyna speciosa Korth) decoction in rats

Hassan Z, Singh D, Suhaimi FW, Chear NJ, Harun N, See CP, et al.

Regul Toxicol Pharmacol, 2023 Sep;143:105466.
PMID: 37536550 DOI: 10.1016/j.yrtph.2023.105466

Mitragyna speciosa Korth also known as kratom, is an herbal drug preparation for its therapeutic properties and opioid-replacement therapy. Kratom is consumed in a brewed decoction form in Malaysia and to date, no studies have characterized its chemical and toxicity profile. Thus, this study aims to evaluate kratom decoction's safety and toxicity profile after 28 days of treatment. Mitragynine content was quantified in kratom decoction and used as a marker to determine the concentration. Male and female Sprague Dawley rats were orally treated with vehicle or kratom decoction (10, 50 or 150 mg/kg) and two satellite groups were treated with vehicle and kratom decoction (150 mg/kg). Blood and organs were collected for hematology, biochemical and histopathology analysis at the end of treatment. No mortality was found after 28 days of treatment and no significant changes in body weight and hematology profile, except for low platelet count. High amounts of uric acid, AST, ALT and alkaline phosphatase were found in the biochemical analysis. Histological investigation of the heart and lungs detected no alterations except for the kidney, liver and brain tissues. In conclusion, repeated administration of kratom decoction provided some evidence of toxicity in the kidney and liver with no occurrence of mortality.
Fulltext Evaluation of the antinociceptive activity and acute oral toxicity of standardized ethanolic extract of the rhizome of Curcuma xanthorrhiza Roxb

Devaraj S, Esfahani AS, Ismail S, Ramanathan S, Yam MF

Molecules, 2010 Apr;15(4):2925-34.
PMID: 20428088 DOI: 10.3390/molecules15042925

Ethanolic extract of Curcuma xanthorrhiza was used to evaluate the analgesic and toxicity effects in vivo. The extract was standardized using GC-MS, which showed that 1 mg of Curcuma xanthorrhiza ethanolic extract contains 0.1238 mg of xanthorrhizol. The analgesic activity was studied in rats using three different models, namely the hot plate test, tail flick test and formalin-induced pain test. The acute oral toxicity was examined by the oral administration of standardized Curcuma xanthorrhiza ethanolic extract in mice at doses ranging from 300-5,000 mg/kg and observation for 14 days. Standardized Curcuma xanthorrhiza ethanolic extract did not show significant analgesic effect in the hot plate and tail flick tests. However, in the formalin-induced pain test, Curcuma xanthorrhiza ethanolic extract significantly (P < 0.05) suppressed the paw licking time of rats in both early and late phases at doses 200 and 400 mg/kg of the extract, respectively. In the acute oral toxicity study, Curcuma xanthorrhiza ethanolic extract did not show any toxic effects in mice at 5 g/kg. These experimental results suggest that the standardized Curcuma xanthorrhiza ethanolic extract showed peripheral and central antinociceptive activity associated with neurogenic pain as well as a relative absence of toxic effects which could compromise the medicinal use of this plant in folk medicine.
Fulltext Evaluation of antioxidant and antibacterial activities of aqueous, methanolic and alkaloid extracts from Mitragyna speciosa (Rubiaceae family) leaves

Parthasarathy S, Bin Azizi J, Ramanathan S, Ismail S, Sasidharan S, Said MI, et al.

Molecules, 2009;14(10):3964-74.
PMID: 19924042 DOI: 10.3390/molecules14103964

Studies on the antioxidant and antimicrobial activities of Mitragyna speciosa leaf extracts are lacking. In this study the antioxidant properties of water, methanolic and alkaloid M. speciosa leaf extracts were evaluated using the DPPH (2,2-diphenyl-1- picrylhydrazyl) radical scavenging method. The amount of total phenolics and flavanoid contents were also estimated. The DPPH IC(50) values of the aqueous, alkaloid and methanolic extracts were 213.4, 104.81 and 37.08 microg/mL, respectively. The total phenolic content of the aqueous, alkaloid and methanolic extracts were 66.0 mg, 88.4, 105.6 mg GAE/g, respectively, while the total flavanoid were 28.2, 20.0 and 91.1 mg CAE/g respectively. The antioxidant activities were correlated with the total phenolic content. This result suggests that the relatively high antioxidant activity of the methanolic extract compared to aqueous and alkaloid extract could be possibly be due to its high phenolic content. The aqueous, alkaloid and methanolic extracts were screened for antimicrobial activity. The extracts showed antimicrobial activity against Salmonella typhi and Bacillus subtilis. The minimum inhibitory concentrations (MICs) of extracts determined by the broth dilution method ranged from 3.12 to 6.25 mg/mL. The alkaloid extract was found to be most effective against all of the tested organisms.

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