Cellular toxicity and DNA damage induced by Newbouldia laevis used for male infertility treatment in prokaryotic and eukaryotic models.

Leaves of Newbouldia laevis have been extensively used in solving problems associated with infertility and childbirth in many African countries. Yet, information is very limited on the DNA damaging potential of this plant. This study evaluated the cytogenotoxic effect of the aqueous extract of N. laevis leaf using prokaryotic models (Ames Salmonella fluctuation test using TA100 and TA98 strains of Salmonella typhimurium and SOS Chromotest with Escherichia coli PQ37) and eukaryotic model (Allium cepa root cells). Identification of the volatile organic compounds (VOCs) and phytochemical screening of the plant extract were also performed. Onion bulbs were grown on each concentration (1 to 50%; v/v, extract/tap water) of the extract for chromosomal aberrations and root growth analyses. Results of the Ames test indicated that the extract is mutagenic while the SOS Chromotest results showed good complementation to the Ames test results, although the E. coli PQ37 system showed slightly higher sensitivity in the detection of mutagenicity and genotoxicity of the extract. The plant extract was cytotoxic when compared to the control, inducing a significant (p < 0.05) concentration-dependent inhibition of root growth from 5 to 50% concentrations. At 50% concentration, the extract completely inhibited cell division in the A. cepa. Also, chromosomal aberration increased significantly (p < 0.05) in exposed onions from 5 to 20% concentrations. The mutagenicity and cytogenotoxicity recorded in this report were believed to be caused by the presence of VOCs such as 1,2,3-benzene-triol, 1,2-benzenediol, and 5-hydroxymethylfurfural, and alkaloids in the extract an indication of the cytogenotoxicity of the aqueous extract of N. laevis leaf even at low concentration.

Effective removal of malachite green oxalate from aqueous solution using Newbouldia laevis husk/MWCNTs nanocomposite: equilibrium, kinetics, and thermodynamics.

The discharge of colored effluent into water bodies is a big concern; hence, the current work was designed to fabricate a superior nanocomposite (NBM) using the Newbouldia laevis husk (NB) and functionalized multiwalled carbon nanotubes (f-MWCNTs) for the adsorption of malachite green oxalate (MGO). Brunauer-Emmett-Teller (BET) surface analysis was used to assess the specific surface area of NB (0.7699 m2 g-1) and NBM (94.006 m2 g-1). Fourier transform infrared spectroscopy (FTIR) was employed to determine the chemical moieties on the surface of the adsorbent. Field emission scanning electron microscopy (FESEM) and thermogravimetric analysis (TGA) were used to analyze the surface morphology and the thermal behavior of the adsorbents. Essential factors of the adsorption process were investigated, and it was revealed that pH 6.0, adsorbent dose of 0.05 g, contact time 80 min, concentration of 100 mg dm-3 and maximum adsorption capacity of 35.78 mg g-1 (NB) and 69.97 mg g-1 (NBM) were the optimal parameters. The NB and NBM adsorption processes followed a pseudo-first-order kinetic model. The exothermic and endothermic adsorptive processes were noticed to be the best descriptions of MGO elimination by NB and NBM, respectively. The uptake of MGO by NB and NBM was best described by models of Freundlich and Langmuir isotherms. Besides, NBM demonstrated uptake efficiency that is >80% after the fourth adsorption/desorption cycle. As a result, NBM has a wide range of possible uses in environmental remediation.

The husk of Newbouldia laevis is a frequent waste that must be managed properly. This paper describes the application of Newbouldia laevis husk as a value-added material for the design of a water treatment agent. The use of carbon nanotube in the modification of Newbouldia laevis husk would have a synergistic effect on the overall property of the nanocomposite. Nanocomposite synthesized from multiwalled carbon nanotubes (MWCNTs) and Newbouldia laevis husk were characterized and used for the sequestration of malachite green oxalate from contaminated water. Our primary goal is to optimize the nanocomposite by varying factors of adsorption such as solution pH, equilibrium, kinetic, thermodynamic, and regeneration studies. We believe that this study will contribute to the existing knowledge of Newbouldia laevis husk. Owing to the exceptional potential of the nanocomposite, this adsorbent can be extended to possible field applications.

Pharmacognostic profiles, evaluation of analgesic, anti-inflammatory and anticonvulsant activities of Newbouldia laevis (P. Beauv.) Seem. ex Bureau leaf and root extracts in Wistar rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Newbouldia laevis is a popular medicinal plant whose leaves and roots are used in Nigeria as ethnomedicinal prescriptions for pain, inflammation, convulsion, and epilepsy. These claims have not been scientifically verified prior to this study. AIM OF THE STUDY: To determine pharmacognostic profiles of the leaves and roots and evaluate the analgesic, anti-inflammatory, and anticonvulsant activities of methanol leaf and root extracts in Wistar rats. MATERIAL AND METHODS: The pharmacognostic profiles of the leaves and roots were determined using standard procedures to serve as fingerprints for the plant. The methanol leaf and root extracts of Newbouldia laevis were tested for acute toxicity using the OECD's up and down method at the maximum dose of 2000 mg/kg (orally) in Wistar rats. Analgesic studies were carried out in acetic acid-induced writhing in rats and tail immersion. The anti-inflammatory activity of the extracts was evaluated using carrageenan-induced rat paw-oedema and formalin-induced inflammation in rats' mode. The anticonvulsant activity was determined using strychnine-induced, pentylenetetrazol-induced, and maximal electroshock-induced rat convulsion models. For each of these studies, the extracts doses of 100, 200 and 400 mg/kg were administered to the rats following the oral route. RESULTS: The pharmacognostic profiles showed that the leaves possessed deep-sunken paracytic stomata (5-8-16 mm2; adaxial, 8-11-24 mm2; abaxial epidermis), vein islets (2-4-10 mm2; adaxial), vein terminations (10-14-18 mm2; adaxial), palisade ratio (8.3-12.5-16.4 mm2; adaxial, 2.5-6.8-12.2 mm2; adaxial), covering unicellular trichome (8-14; adaxial), spheroidal calcium oxalate crystals (3-5 µm), and oval-shaped striated starch grain with no hilum (0.5-4.3 µm). The transverse section of the leaf showed the presence of spongy and palisade parenchyma as well as a closed vascular bundle. The root powder showed the presence of brachy sclereid, fibers without lumen, and lignin. All physicochemical parameters fall within the acceptable limits, phytochemical contents showed mainly glycosides, alkaloids, and steroids while acute oral toxicity (LD50) of the parts for 14 days did not produce any toxicity signs or mortality in the rats. The extracts produced dose-dependent (100-400 mg/kg) analgesic involving opioid receptors, anti-inflammatory, and anticonvulsant activities in the rats which were significant (p ≤ 0.05) when compared to the standard drugs. The leaf extract possessed the most potent analgesic and anti-inflammatory effects in the rats, while the most anticonvulsant effects were observed in rats treated with the leaf extract. Both extracts showed elevated levels of protection against strychnine-induced, pentylenetetrazol-induced, and maximal electroshock-induced seizure in rats. CONCLUSION: Our study revealed some pharmacognostic profiles of Newbouldia laevis leaves and roots that are vital for its identification from closely related species often used for adulteration in traditional medicine. The study further showed that the leaf and root extracts of the plant possessed dose-dependent analgesics, anti-inflammatory and anti-convulsant activities in rats, thus, justifying its use for the treatment of these diseases in Nigerian traditional medicine. There is a need to further study its mechanisms of action towards drug discovery.

Newboulasides A and B, two new caffeic acid glycosides from Newbouldia laevis with α-amylase inhibitory activity.

Chemical investigation of the ethanol extract of the leaves of Newbouldia laevis (P. Beauv) led to the isolation of two new caffeic acid glycosides, Newboulasides A (1) and B (2). The structures of these compounds were determined on the basis of extensive spectroscopic methods, including 1D-, 2D-NMR and MS data. The extracts and fractions and the isolated compounds were evaluated for their inhibition of α-amylase enzyme activity. The extract showed inhibition of α-amylase activity with IC50 value of 102.91 µg/mL, while the isolated compounds (1 and 2) exhibited pronounced inhibition with IC50 values of 4.95 and 4.44 µg/mL respectively, comparable to the standard - Acarbose with IC50 value of 4.05 µg/mL. Our findings demonstrated that the inhibition of α-amylase activity may be part of the mechanisms through which N. leavis exhibits antidiabetic effect.

Secondary metabolites of endophytic fungi from Newbouldia laevis and Cassia tora leaves: prospecting for new antimicrobial agents.

AIM: This study tries to prospect for new antimicrobial agents using some Nigerian plants Background: Antimicrobial compounds from fungi endophytes have shown great promise in mitigating the threats of resistant pathogens. OBJECTIVE: The study evaluated the in vitro antimicrobial property of secondary metabolites of endophytic fungi isolated from Newbouldia laevis and Cassia tora leaves. METHODS: Ten endophytic fungi were isolated from the two plants' leaves and later fermented on local rice for 21 days. Thereafter, their secondary metabolites were extracted using ethyl acetate. The antibacterial activity of the extracts on the test organisms were determined using agar diffusion and agar dilution methods, while the bioactive constituents were identified using High performance liquid chromatography coupled to diode array detector. RESULTS: Nine of the crude extracts (NL1, NL3, NL6, NL10, NL12, CT2, CT7, CT9 and CT10) of the fungi isolates inhibited at least one of the microorganisms studied with maximum and minimum Inhibition-Zone-Diameter of 14 mm and 2 mm respectively while CT1 did not inhibit any of the tested microorganisms at tested concentrations. The extracts exhibited good antifungal activity, inhibiting the growth of both C. albicans and Trichophyton tested with an InhibitionZone-Diameter ranging between 4-8 mm and 7-14 mm respectively. The endophytic fungi extracts- CT2 and NL1- exhibited the best antimicrobial activity, inhibiting most of the tested microorganisms. HPLC-DAD analysis of the endophytic fungal extracts identified some classes of compounds such as catechin derivatives, benzoic acid derivatives and apigenin, which were previously reported to have antimicrobial potentials. CONCLUSION: Newbouldia laevis and Cassia tora leaves house endophytic fungi capable of yielding secondary metabolites with potential as anti-infective agents.

Newbouldia laevis improved glucose and fat homeostasis in a TYPE-2 diabesity mice model.

ETHNOPHARMACOLOGICAL RELEVANCE: Newbouldia laevis (P. Beauv) has a long-standing ethnomedicinal use in the management of diabetes mellitus. However, no scientific evidence has established its potentials in the management of obesity dependent diabetes. AIM: This study aimed at filling the gap in knowledge about the antidiabetic activity of the leaf extract of N. laevis in a type-2 diabesity mice model. MATERIALS AND METHOD: The ethanol leaf extract was subjected to liquid-liquid partitioning successively with n-hexane, ethyl acetate and butanol to obtain respective fractions soluble in these solvents. The butanol (most active) fraction at 302 and 604 mg/kg was further tested on high-fat diet STZ-NAD induced type-2 diabetic mice for 10 weeks with glibenclamide (10 mg/kg) and pioglitazone (30 mg/kg) as standards. The effect on food intake, body weight, fasting blood glucose, oral glucose tolerance test (OGTT), oral fat tolerance test (OFTT), insulin secretion, insulin resistance and lipid profile were determined prior to treatment, mid-way and at the end of 10 weeks treatment. RESULTS: Higher food consumption was recorded in diabetic (D) animals on high-fat diet (HFD) compared to the normal diet (ND)-fed groups. Treatment of these diabetic mice on HFD with 604 mg/kg of butanol fraction produced significant (p < 0.05) reduction in body weights of these animals from the 2nd week to the 9th week. Ten weeks treatment with butanol fraction achieved a marked decrease in blood glucose and also an increase in fat clearance. Just like pioglitazone, treatment with butanol fraction at both 302 and 604 mg/kg doses produced significant (p < 0.05) decrease in HFD mediated elevation of serum insulin and a non-significant (p > 0.05) increase in STZ-NAD mediated depletion of serum insulin. Butanol fraction at 604 mg/kg also produced reduction in insulin resistance as indicated by significant (p < 0.05) decrease in HOMA-IR value on the 5th and 10th week just like pioglitazone (30 mg/kg). CONCLUSION: N. laevis exhibited wide actions in the regulation of glucose and fat homeostasis making it a potential novel agent for the management of diabetes, obesity and their likely associated complications.

Anxiolytic- and antidepressant-like activities of hydroethanol leaf extract of Newbouldia laevis (P.Beauv.) Seem. (Bignoniaceae) in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Newbouldia laevis (Bignoniaceae) is a woody tropical plant commonly found in southwest Nigeria. Ethnobotanical survey and literature revealed its application in the management of CNS disorders e.g. psychosis, insomnia, convulsions and associated anxiety and depression. This study evaluated the anxiolytic and antidepressant activities of the hydroethanol leaf extract of N. laevis in mice. MATERIALS AND METHODS: The hole-board (HBT), elevated plus maze (EPMT), light/dark exploration (LDET), open field (OFT), social interaction (SIT) (anxiolytic activity), forced swim (FST) and tail suspension (TST) (antidepressant property) tests were employed in this investigation. Mice randomly allotted to different groups were treated orally with distilled water (10 ml/kg), diazepam (1 and 3 mg/kg), imipramine (20 mg/kg) and N. laevis (25-200 mg/kg). The mice were subjected to the various tests 60 min post-treatment. RESULTS: In the HBT, N. laevis (25 and 100 mg/kg) increased the number of sectional crossings significantly (p < 0.05). In the OFT, N. laevis (25-200 mg/kg) increased the number of general square crossings, centre square crossings, rearings and assisted rearings (p < 0.05). In the EPMT, the extract (25 and 50 mg/kg) increased the open arms time spent, number of head dips and entry (p < 0.05). In the LDET, N. laevis increased the number of transitions at 100 and 200 mg/kg (p < 0.05). For the SIT, N. laevis (25 and 200 mg/kg) increased the frequency and duration of interaction respectively (p < 0.05). In the FST and TST, N. laevis (25-200 mg/kg) increased the latency and reduced the total duration of immobility (p < 0.05). The effect of the extract on duration of immobility was significantly reversed by sulpiride (Dopamine D2 receptor antagonist). CONCLUSION: The hydroethanol leaf extract of N. laevis possesses anxiolytic- and antidepressant-like activities, the later possibly mediated by dopaminergic enhancement(s).

In Vitro Reversible and Time-Dependent CYP450 Inhibition Profiles of Medicinal Herbal Plant Extracts Newbouldia laevis and Cassia abbreviata: Implications for Herb-Drug Interactions.

This study evaluated the effects of Newbouldia laevis and Cassia abbreviata extracts on CYP450 enzyme activity. Recombinant CYP450 enzyme and fluorogenic substrates were used for evaluating inhibition, allowing the assessment of herb-drug interactions (HDI). Phytochemical fingerprinting was performed using UPLC-MS. The herbal extracts were risk ranked for HDI based on the IC50 values determined for each CYP enzyme. Newbouldia laevis inhibited CYP1A2, CYP2C9, and CYP2C19 enzyme activities with Ki of 2.84 µg/mL, 1.55 µg/mL, and 1.23 µg/mL, respectively. N. laevis exhibited a TDI (4.17) effect on CYP1A2 but not CYP2C9 and CYP2C19 enzyme activities. Cassia abbreviata inhibited CYP1A2, CYP2C9, and CYP2C19 enzyme activities showing a Ki of 4.86 µg/mL, 5.98 µg/mL, and 1.58 µg/mL, respectively. TDI potency assessment for Cassia abbreviata showed it as a potential TDI candidate (1.64) for CYP1A2 and CYP2C19 (1.72). UPLC-MS analysis showed that Newbouldia laevis and Cassia abbreviata possess polyphenols that likely give them their therapeutic properties; some of them are likely to be responsible for the observed inhibition. The observations made in this study suggest the potential for these herbal compounds to interact, especially when co-administered with other medications metabolized by these CYP450 enzymes.

In vitro antioxidant capacity and free radical scavenging evaluation of active metabolite constituents of Newbouldia laevis ethanolic leaf extract

BACKGROUND: The aim of the present study was to evaluate the in vitro antioxidant and free radical scavenging capacity of bioactive metabolites present in Newbouldia laevis leaf extract. RESULTS: Chromatographic and spectrophotometric methods were used in the study and modified where necessary in the study. Bioactivity of the extract was determined at 10 µg/ml, 50 µg/ml, 100 µg/ml, 200 µg/ml and 400 µg/ml concentrations expressed in % inhibition. The yield of the ethanolic leaf extract of N.laevis was 30.3 g (9.93%). Evaluation of bioactive metabolic constituents gave high levels of ascorbic acid (515.53 ± 12 IU/100 g [25.7 mg/100 g]), vitamin E (26.46 ± 1.08 IU/100 g), saponins (6.2 ± 0.10), alkaloids (2.20 ± 0.03), cardiac glycosides(1.48 ± 0.22), amino acids and steroids (8.01 ± 0.04) measured in mg/100 g dry weight; moderate levels of vitamin A (188.28 ± 6.19 IU/100 g), tannins (0.09 ± 0.30), terpenoids (3.42 ± 0.67); low level of flavonoids (1.01 ± 0.34 mg/100 g) and absence of cyanogenic glycosides, carboxylic acids and aldehydes/ketones. The extracts percentage inhibition of DPPH, hydroxyl radical (OH.), superoxide anion (O2 .-), iron chelating, nitric oxide radical (NO), peroxynitrite (ONOO-), singlet oxygen (1O2), hypochlorous acid (HOCl), lipid peroxidation (LPO) and FRAP showed a concentration-dependent antioxidant activity with no significant difference with the controls. Though, IC50 of the extract showed significant difference only in singlet oxygen (1O2) and iron chelating activity when compared with the controls. CONCLUSIONS: The extract is a potential source of antioxidants/free radical scavengers having important metabolites which maybe linked to its ethno-medicinal use.

Uterine contractile effects of the aqueous and ethanol leaf extracts of Newbouldia Laevis (Bignoniaceae) in vitro.

Based on traditional reports, the aqueous and ethanol extracts of the leaves of N. laevis were tested on isolated uterine preparations of non-pregnant rats. The effects of increasing cumulative concentrations of the extracts on the amplitude and frequency of spontaneously contracting uterine tissues were tested. Direct effects of the extracts and acetylcholine on uterine smooth muscle were also tested in organ baths containing aerated physiological salt solution maintained at 37 degrees . The EC(50) and Emax were determined and analyzed using one way ANOVA with Dunnett's post hoc test. The extract significantly increased the frequency (P<0.05) of spontaneous contractions without significantly affecting the amplitude. The extracts and acetylcholine were observed to directly stimulate uterine contractions, however there were significant differences (P<0.05) in their EC(50) and Emax. In conclusion, the leaves of N. laevis increase the frequency of spontaneously contracting tissues and directly stimulate uterine contractions which may account for the use of the leaf extract traditionally.