Marco Antonio Botelho

Sida cordifolia Leaf Extract Reduces the Orofacial Nociceptive Response in Mice

In this study, we describe the antinociceptive activity of the ethanol extract (EE), chloroform (CF) and methanol (MF) fractions obtained from Sida cordifolia, popularly known in Brazil as “malva branca” or “malva branca sedosa”. Leaves of S. cordifolia were used to produce the crude ethanol extract and after CF and MF. Experiments were conducted on Swiss mice using the glutamate and formalin‐induced orofacial nociception. In the formalin test, all doses of EE, CF and MF significantly reduced the orofacial nociception in the first (p < 0.001) and second phase (p < 0.001), which was also naloxone‐sensitive. In the glutamate‐induced nociception test, only CF and MF significantly reduced the orofacial nociceptive behavior with inhibition percentage values of 48.1% (100 mg/kg, CF), 56.1% (200 mg/kg, CF), 66.4% (400 mg/kg, CF), 48.2 (200 mg/kg, MF) and 60.1 (400 mg/kg, MF). Furthermore, treatment of the animals with EE, CF and MF was not able to promote motor activity changes. These data demonstrate that S. cordifolia has a pronounced antinociceptive activity on orofacial nociception. However, pharmacological and chemical studies are necessary in order to characterize the responsible mechanisms for this antinociceptive action and also to identify other bioactive compounds present in S. cordifolia. Copyright

Citral reduces nociceptive and inflammatory response in rodents

Citral (CIT), which contains the chiral enantiomers, neral (cis) and geranial (trans), is the majority monoterpene from Lippia alba and Cymbopogon citratus. The present study aimed to evaluate CIT for antinociceptive and anti-inflammatory activities in rodents. Antinociceptive and anti-inflammatory effects were studied by measuring nociception through acetic acid and formalin tests, while inflammation was verified by inducing peritonitis and paw edema with carrageenan. All tested doses of CIT had significant protection (p<0.001) against acetic acid (0.8%) induced nociceptive behavior and the effects were also similar to morphine while formalin induced nociception was significantly protected (p<0.05) only at higher dose (200 mg/kg) of CIT in the first phase of the test. CIT significantly reduce (p<0.001) nociceptive behavior emanating from inflammation in second phase at all the doses.The pretreatment with CIT (100 and 200 mg/kg) significantly reduced the paw edema induced by carrageenan. Moreover, systemic treatment with CIT (100 and 200 mg/kg) significantly reduced (p<0.001) the leukocyte migration in the carrageenan-induced migration to the peritoneal cavity. Our investigation shows that CIT possess significant central and peripheral antinociceptive effects. It was also verified an anti-inflammatory activity. All together these results suggest that CIT might represent important tool for treatment of painful conditions.

Bioassay‐guided Evaluation of Antinociceptive Properties and Chemical Variability of the Essential Oil of Hyptis fruticosa

The composition of three samples of essential oil (EO) extracted from the leaves and flowers of Hyptis fruticosa (Lamiaceae) were investigated by GC/MS and GC‐FID. The variability of the constituents and biological activity were evaluated in the oil samples. Acetic acid‐induced abdominal constrictions and formalin‐induced pain tests in mice were used for screening the antinociceptive activity. The possible antagonism of the essential oils or morphine (MOR) antinociceptive effects by pretreatment with naloxone, showed no influence on the antinociceptive action of the oils in the acetic acid‐induced writhing test. All examined oil samples presented antinociceptive activity. The oil sample obtained from the leaves collected during the vegetative growth stage, near São Cristóvão at Sítio Tujubeba exhibited the highest effect. The same oil sample had a main percentage of 1,8‐cineole (18.70%). Nevertheless, the oil obtained from flowers collected at the same location, showed a significant difference (p < 0.05) in the response intensity in the first phase of paw licking (100 mg/kg) possibly due to the higher contents of α‐pinene (20.51%) and β‐pinene (13.64%). The results provide evidence for the use of H. fruticosa by traditional medicine practitioners in the management of pain. Copyright

Hyptis pectinata: Redox Protection and Orofacial Antinociception

Hyptis pectinata L. Poit, known as ‘sambacaitá’, is used in Brazil to treat inflammatory and painful disorders. In this study, the antioxidant and orofacial antinociceptive properties of the aqueous extract of H. pectinata leaves (AEPH) were assessed using in vitro and in vivo models. Thus, AEPH reduced the 2,2‐diphenyl‐1‐picrylhydrazyl radical up to 72.10% with an EC50 of 14.56 µg/ml. It also inhibited 40.80% of the lipoperoxidation induced by 2′‐azobis (2‐amidinopropane) dihydrochloride in the thiobarbituric acid‐reactive substances assay. The orofacial antinociceptive activity was evaluated in mice pre‐treated with AEPH (100, 200 and 400 mg/kg, p.o.) and morphine (5 mg/kg, i.p.), which received afterwards formalin‐ (20 µl, 2% solution, s.c.), glutamate‐ (40 µl, 25 mM, s.c.) and capsaicin‐ (20 µl, 2.5 µg, s.c.) to induce orofacial nociception. AEPH at all doses reduced (p < 0.001) the nociceptive response in the first (43–62%) and second (47–80%) phases of the formalin test. Besides, the effect of AEPH (400 mg/kg) was not changed in the presence of naloxone (1.5 mg/kg, i.p.), an opioid antagonist. AEPH significantly inhibited mice face rubbing for capsaicin (23–69%, p < 0.05) and glutamate (48–77%, p < 0.001) at all doses. The findings suggested the AEPH has peripheral and central antinociceptive activities, which are not related to opioid receptors. Copyright

Phythochemical screening and antimicrobial activity phythochemical of essential oil from Lippia gracillis

The chemical composition of the essential oil obtained from the fresh and dried leaves of Lippia gracillis Schauer, Verbenaceae, was analyzed by gas chromatography and gas chromatography/mass spectrometry (GC⁄MS). The yield of essential oil extracted from the dried leaves was significantly higher (p<0.05) when compared to the fresh leaves. Seventeen components were identified. The monoterpenes and sesquiterpene hydrocarbons with 96.26% (w/w) of the total oil obtained of fresh leaves and 86.99% (w/w) of the total oil obtained of dried leaves were the principal compound groups. Thymol was observed dominant (44.42%; 21.3%), followed by carvacrol (22.21%; 21.30%), p-cymene (6.23%; 8.58%), α-pinene (5.65%; 19.42%), β-caryophyllene (5.61%; 3.57%) and other minor constitutes, respectively. Microbiological results obtained by agar diffusion method, micro dilution method and minimum inhibitory concentration (MIC) showed that the essential oil has a relevant antimicrobial activity against E. coli (ATCC 10536), E. coli (Ec 27), Pseudomonas aeruginosa (ATCC 15442), S. aureus (ATCC 12692) and S. aureus (Sa 358), with their inhibition zones ranging from 9 to 13 mm and the MIC ranging from 64 to 512 μg/mL.

Nanostructured transdermal hormone replacement therapy for relieving menopausal symptoms: a confocal Raman spectroscopy study

OBJECTIVE: To determine the safety and efficacy of a transdermal nanostructured formulation of progesterone (10%) combined with estriol (0.1%) + estradiol (0.25%) for relieving postmenopausal symptoms. METHODS: A total of 66 postmenopausal Brazilian women with climacteric symptoms of natural menopause received transdermal nanostructured formulations of progesterone and estrogens in the forearm daily for 60 months to mimic the normal ovarian secretory pattern. Confocal Raman spectroscopy of hormones in skin layers was performed. Clinical parameters, serum concentrations of estradiol and follicle-stimulating hormone, blood pressure, BI-RADS classification from bilateral mammography, and symptomatic relief were compared between baseline and 60 months post-treatment. Clinicaltrials.gov: NCT02033512. RESULTS: An improvement in climacteric symptoms was reported in 92.5% of women evaluated before and after 60 months of treatment. The serum concentrations of estradiol and follicle-stimulating hormone changed significantly (p<0.05) after treatment; the values of serum follicle-stimulating hormone decreased after 60 months from 82.04±4.9 to 57.12±4.1 IU/mL. A bilateral mammography assessment of the breasts revealed normal results in all women. No adverse health-related events were attributed to this hormone replacement therapy protocol. CONCLUSION: The nanostructured formulation is safe and effective in re-establishing optimal serum levels of estradiol and follicle-stimulating hormone and relieving the symptoms of menopause. This transdermal hormone replacement therapy may alleviate climacteric symptoms in postmenopausal women.

EFFECTS OF A NEW TESTOSTERONE TRANSDERMAL DELIVERY SYSTEM, BIOLIPID B2®-TESTOSTERONE IN HEALTHY MIDDLE AGED MEN: A CONFOCAL RAMAN SPECTROSCOPY STUDY

The Transdermal Delivery System (BIOLIPID B2®) is a nanoemulsion that can be a pplied directly to the skin by a metered pump to deliver hormones into the systemic circulation. The ai ms of this study were to assess the efficacy of BIOLIPID B2 ® to deliver testosterone systemically and evaluate the short - term effects on Thyroid - Stimulating Hormone (TSH), C - reactive protein (CRP) serum levels and blood pressure in middle aged men with tes tosterone deficiency . An open label randomized prospective study enrolled 14 Brazilian middle aged men with testosterone deficiency . The volunteers received in the forearm 50mg of transdermal BIOLIPID B2® - testosterone nanostructured formulation daily for 3 months. Confocal Raman Spectroscopy (CRS) was used to determine depth and concentration of testosterone on skin layers. Clinical parameters and serum concentrations were compared between the base line and 3 months after treatment. Serum concentrations of t estosterone, CPR and TSH changed significantly (p 0.05) and no changes were observed in the blood pressure . N o adverse events were attribu ted to this transdermal hormone replacement therapy protocol. The BIOLIPID B2 ® nanoemulsion formulation is safe and effective in reestabli shing testosterone and Thyroid - Stimulating Hormone (T SH) serum levels. This transdermal protocol may serve as a strate gy for hormone replacement therapy in middle aged men.

Chemical composition of the essential oil of Lippia gracilis Schauer leaves and its potential as modulator of bacterial resistance

Lippia gracilis Schauer (Verbenaceae) is an aromatic plant widely used in folk medicine in the Brazilian semi-arid region. This work aims at evaluating the anti-microbial activity of essential oils from the leaves of L. gracilis (EOLG) when associated with commercial antibiotics against bacterial resistance. The essential oil obtained by hydrodistillation has 38 chemical constituents with thymol (44.4%), carvacrol (22.2%), p-cymene (6.2%) and α-pinene (5.6%) as major constituents. Microbiological assays were conducted by direct and gaseous contacts. We demonstrated that EOLG has significant synergism with all antibiotics tested. EOLG decreased the minimal inhibitory concentration of the aminoglycosides and improved their zone of inhibition more than 300% against Staphylococcusaureus and 270% against Pseudomonasaeruginosa. These results demonstrate that natural substances present in EOLG have the capacity to modulate bacterial resistance and can be used as an adjuvant therapy against multi-resistant microorganisms such as S. aureus, Escherichia coli and P. aeruginosa.

Effect of Collection Time on Composition of Essential Oil of Lippia gracilis Schauer (Verbenaceae) Growing in Northeast Brazil

Abstract The specie Lippia gracilis Schauer is largely utilized for their antimicrobial properties, which are attributed to the presence of phenolic monoterpenes such as thymol and carvacrol. The aim of this study was to determine the yields and chemical composition of the essential oils obtained from fresh and dry leaves of collected at different times in the city of Crato-CE, in January 2009. The essential oils were extracted by hydrodistillation, and its components were identified by gas chromatography coupled with mass spectrophotometry. The time of plant collection should coincide with the moment of greatest expression of the active principle.

Enhancement of the antibiotic activity of erythromycin by volatile compounds of Lippia alba (Mill.) N.E. Brown against Staphylococcus aureus.

Background: Lippia alba (Mill.) N.E. Brown, popularly known as “erva-cidreira,” is commonly found in northeastern Brazil. The leaves tea is used to treat digestive disturbances, nausea, cough, and bronchitis. Objective: This work reports the chemical composition and erythromycin-modifying activity by gaseous contact against Staphylococcus aureus. Materials and Methods: The leaves of L. alba were subjected to hydrodistillation, and the essential oil extracted was examined with respect to the chemical composition, by gas chromatography-mass spectrometry (GC-MS), and the essential oil extracted was evaluated for antibacterial and antibiotic-modifying activity by gaseous contact. Results: The overall yield of essential oil obtained by hydrodistillation was 0.52%. The GC-MS analysis has led to the identification of the main components: geranial (31.4%) and neral (29.5%). It was verified that the essential oil interfered with erythromycin antibiotic activity against S. aureus ATCC 25923 was enhanced (221.4%) in the presence of 12% essential oil. The 3% essential oil increased the effect against S. aureus ATCC 25923 (41.6%) and S. aureus ATCC 6538 (58.3%). Conclusion: The essential oil of L. alba influences the activity of erythromycin and may be used as an adjuvant in antibiotic therapy against respiratory tract bacterial pathogens. Conclusion: The essential oil of L. alba influences the activity of erythromycin and may be used as an adjuvant in antibiotic therapy against respiratory tract bacterial pathogens.