In Vitro Potential of the Acetone Leaf Extract and Fractions of Psychotria capensis (Eckl.) Vatke (Rubiaceae) to Combat Co-Infection of Tuberculosis and Helminthiasis

Tuberculosis (TB) is a disease of global importance that affects millions of people. Approximately a quarter of the world’s population is currently infected with M. tuberculosis, and about 10% of those infected will develop into active disease, particularly immune compromised individuals. Helminthiasis is of global health importance, affecting over 2 billion people mostly in resource-poor countries. Co-infection with tuberculosis (TB) and helminths (worms) is an emerging global public health concern with both affecting about one-third of the global population. Chronic infection with helminths can result in impaired immune responses to TB as well as enhancing failure to TB therapy and BCG vaccination. Antimycobacterial and anthelmintic activities of the acetone extract and fractions of Psychotria capensis were evaluated, including their in vitro safety. In addition, the anti-inflammatory and immunomodulatory effect of the fractions and crude extract of P. capensis were assessed. Antimycobacterial activity of the extract and fractions was tested against four non-tuberculous mycobacteria (Mycobacterium smegmatis, M. fortuitum, M. aurum, M. bovis BCG) and pathogenic M. tuberculosis H37Rv while the Egg Hatch Assay (EHA) was used for the anthelmintic test on eggs of Haemonchus contortus. Cytotoxicity was determined against Vero kidney cells while in vitro immune modulation via cytokine production was determined on activated macrophages. The minimum inhibitory concentration (MIC) values of the Psychotria capensis acetone extract and fractions ranged from 39 to 1,250 μg/ml with the crude extract and hexane fraction having the best MIC values (both 39 μg/ml). In the EHA, the inhibitory concentration (IC50) ranged from 160 to 630 μg/ml with the hexane fraction having the best activity. The hexane and chloroform fractions were relatively non-toxic with LC50 values of 290 and 248 μg/ml respectively, while the acetone crude extract (64 μg/ml) and n-butanol fraction (71 μg/ml) were moderately toxic. The SI values (LC50/MIC) ranged from 0.1 to 7.4 with the hexane fraction having the highest value against M. smegmatis (7.4). The hexane fraction had the best dual anthelmintic and antimycobacterial activity. This fraction had the best NO inhibitory activity and was the least cytotoxic, indicating that its activity was not due to general metabolic toxicity, with 96.54% cell viability. Pro-inflammatory cytokines such as IL-12p70 were upregulated while IL-10 expression was inhibited by the extracts. Compounds were detected using GC-MS analysis, and in both the crude acetone extract and the hexane fraction was the diterpene neophytadiene, which has anti-inflammatory and antimicrobial activity. Finding alternative or complementary approaches to dealing with TB infections by, amongst other things, reducing the incidence of helminth infestations may lessen the burden of TB, contributing to slowing the spread of multi-drug resistance.

Antimicrobial activity by solvents extracted from Ocimum basilicum herb against multidrug resistant gram negative rods.

ABSTRACT: Background: Failure of treatment with antibiotics occurs due to increase in number of Multidrug resistant gram negative bacteria, worldwide. The objective of this study was to find out the antimicrobial activity of crude ethanolic extract and its further three fractions by Ocimum basilicum leaves against multi drug resistant gram negative rods. Material and Methods: This descriptive study was conducted in the Department of Microbiology, University of Health Sciences, Lahore from 1st july 2016 to 30th june 2017. Total 80 multidrug resistant gram negative rods were included in this study. Agar dilution method was performed to determine MIC of crude ethanolic extract and different fractions i-e n-hexane, chloroform and ethyl acetate of Ocimum basilicum leaves against multidrug resistant gram negative rods i-e ESBLs and carbapenemase producers. Muti-inoculater was used for inoculation. Results: The mean MICs of crude ethanolic extract, n-hexane fraction, chloroform fraction, and ethyl acetate fraction of Ocimum basilicum against ESBLs were 100.0±8.00, 168.13±8.00, 176.88±8.00 41.75±8.00 respectively. Similarly, the mean MICs of crude ethanolic extract, n-hexane fraction, chloroform fraction, and ethyl acetate fraction of Ocimum basilicum against carbapenemase producers were 77.50±8.00, 113.75±8.00, 132.50±8.00 and 29.50±8.00 respectively. Conclusions: Ethyl acetate fraction and crude ethanolic extract from leaves of Ocimum basilicum showed good antibacterial effectiveness against ESBLs and carbapenem resistant organisms than other fractions. This finding may also promote the effective use of O. basilicum herb and its components in modern medicine.

Aktivitas Larvasida Fraksi N-Heksan Ekstrak Etanol Daun Mengkudu (Morinda citrifolia. L) terhadap Larva Aedes sp.

Aedes sp. is a vector of the dengue virus that causes Dengue Hemorrhagic Fever (DHF).Larvicides are the optimal method for controlling mosquito development. Temephos is a larvicidalagent of the organophosphate group which is reported to cause side eff ects and ecological hazards,as well as resistance based on reports in several country. This study aims to determine the larvicidalactivity of the n-hexane fraction of Morinda citrifolia leaf ethanol extract on Aedes sp. The compoundgroups in the fraction were identifi ed using TLC through UV light and spray reagents. There were sixtypes of treatment including four concentration fractions (400, 600, 800, and 1000 ppm) as treatment,positive control (temephos 1%) and negative control 1% acetone solution. Twenty-fi ve mosquito larvaeof Aedes sp. tested for each treatment. Larval mortality was recorded and LC50 and LC99 values wereanalyzed using the probit. The results showed that the TLC test of the n-hexane fraction was positivefor terpenoids, anthraquinones, phenols, tannins, and fl avonoids. The results showed that the LC50 andLC99 values were 1040 ppm and 2439 ppm. Therefore, the n-hexane fraction of the ethanol extract hadlarvicidal activity on Aedes sp with li le toxicity.

GC-MS Analysis and Biomedical Therapy of Oil from n-Hexane Fraction of Scutellaria edelbergii Rech. f.: In Vitro, In Vivo, and In Silico Approach

The current study aimed to explore the crude oils obtained from the n-hexane fraction of Scutellaria edelbergii and further analyzed, for the first time, for their chemical composition, in vitro antibacterial, antifungal, antioxidant, antidiabetic, and in vivo anti-inflammatory, and analgesic activities. For the phytochemical composition, the oils proceeded to gas chromatography-mass spectrometry (GC-MS) analysis and from the resultant chromatogram, 42 bioactive constituents were identified. Among them, the major components were linoleic acid ethyl ester (19.67%) followed by ethyl oleate (18.45%), linolenic acid methyl ester (11.67%), and palmitic acid ethyl ester (11.01%). Tetrazolium 96-well plate MTT assay and agar-well diffusion methods were used to evaluate the isolated oil for its minimum inhibitory concentrations (MIC), minimum bactericidal concentration (MBC), half-maximal inhibitory concentrations (IC50), and zone of inhibitions that could determine the potential antimicrobial efficacy’s. Substantial antibacterial activities were observed against the clinical isolates comprising of three Gram-negative bacteria, viz., Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa, and one Gram-positive bacterial strain, Enterococcus faecalis. The oils were also effective against Candida albicans and Fusarium oxysporum when evaluated for their antifungal potential. Moreover, significant antioxidant potential with IC50 values of 136.4 and 161.5 µg/mL for extracted oil was evaluated through DPPH (1,1-Diphenyl-2-picryl-hydrazyl) and ABTS assays compared with standard ascorbic acid where the IC50 values were 44.49 and 67.78 µg/mL, respectively, against the tested free radicals. The oils was also potent, inhibiting the α-glucosidase (IC50 5.45 ± 0.42 µg/mL) enzyme compared to the standard. Anti-glucosidase potential was visualized through molecular docking simulations where ten compounds of the oil were found to be the leading inhibitors of the selected enzyme based on interactions, binding energy, and binding affinity. The oil was found to be an effective anti-inflammatory (61%) agent compared with diclofenac sodium (70.92%) via the carrageenan-induced assay. An appreciable (48.28%) analgesic activity in correlation with the standard aspirin was observed through the acetic acid-induced writhing bioassay. The oil from the n-hexane fraction of S. edelbergii contained valuable bioactive constituents that can act as in vitro biological and in vivo pharmacological agents. However, further studies are needed to uncover individual responsible compounds of the observed biological potentials which would be helpful in devising novel drugs.

The Concentrations of Cholesterol and Reproduction Hormones in Serum of Rabbits Doe That Consumed Moringa oleifera Leaf Extracts

This study was conducted to examine the effect of various kinds of Moringa leaf extracts on cholesterol, estradiol, FSH, LH levels and litter size in rabbit does. The extracts used were hexane fraction extract (HeEF), ethanol fraction extract (EtEF) and ethanol rough extract (EtCE) from moringa leaf flour. Seven treatments were administered including extracts in pellets at a dose of 0, 0.13% HeEF, 0.26% HeEF, 0.37 EtEF, 0.74% EtEF, 0.54% EtCE and 1.08% EtCE. Each treatment was reduplicated on 4 rabbits. Cholesterol levels, estradiol, LH and FSH blood serum, mating readiness and litter size were measured, which results showed that the treatment significantly (P <5%) lowered the cholesterol level and increased the estradiol level in blood serum. On the other hand, the treatment had no significant effect on FSH, LH and litter size levels except at LH levels 2 hours after mating.

Central nervous system depressant activity of fractions of Globimetula braunii Engl. (Loranthaceae) growing on Terminalia catappa L. (Combretaceae) and isolation of lupeol

Globimetula braunii is a parasitic plant belonging to the family Loranthaceae. Traditionally, the plant has been reported to be used in the treatment of insomnia. The study was carried out to investigate the central nervous system (CNS) depressant activity of the fractions of Globimetula braunii growing on Terminalia catappa and to isolate the phytochemical compound(s) present in the most active fraction. The CNS depressant activity of all the fractions was investigated using diazepam induced sleep. The most active fraction was further subjected to the hole board test and beam walk assay. The chromatographic technique was used for the isolation of phytochemical compound. Hexane fraction significantly (p<0.05) reduced latency to sleep and prolonged the sleeping time. Both chloroform and ethylacetate fractions at highest and median doses showed significant increase in the duration of sleep compared to normal saline. The n-butanol fraction at all doses tested do not have any effect on time of onset and duration of sleep when compared with normal saline treated group. Hexane fraction significantly (p<0.05) decreased the number of head dip in a dose dependent manner and delayed the time to reach the goal box compared to normal saline treated group. Lupeol was isolated from n-hexane fraction.

Uji Perbandingan Aktivitas Penyembuhan Luka Sayat Fraksi n-Heksan, Metanol, dan Ekstrak Daun Talas (Colocasia esculenta (L.) Schott)

Talas or taro leaf contains active compounds in the form of flavonoids and saponins that can accelerate the process of epidermal tissue re-epitalization and infiltration of inflammatory cells in the wound area. The purpose of this study was to compare the healing activities of the n-hexane fraction, methanol fraction, and taro leaf extract in New Zealand rabbits. The extraction method using maceration, the fractionation method used liquid-liquid partition with a separating funnel and the treatment method on test animals used was the application of the n-hexane fraction, methanol fraction, and taro leaf extract on open wounds of New Zealand rabbits with positive control and negative control. The data obtained is in the form of the value of open wound narrowing on the rabbit’s back. The result of the data were analyzed by ANOVA (Analysis Of Variant) followed by the LSD (Least Significant Different) test to see which treatment gave different effect in each group. The results of the research or testing for 14 days showed that the extract, n-hexane fraction, and methanol fraction had wound healing activity on the rabbit back with statistical data showing significant differences between treatment groups. The extract had the most optimal wound healing activity compared to other treatment groups.Keywords: talas leaf, extract, fraction, wound AbstrakDaun talas mengandung senyawa aktif berupa flavonoid dan saponin yang mampu mempercepat proses reepitalisasi jaringan epidermis dan infiltrasi sel-sel radang pada daerah luka. Tujuan dari penelitian ini yaitu mengetahui perbandingan aktivitas penyembuhan luka terbuka fraksi n-heksan, fraksi metanol, dan ekstrak daun talas pada kelinci New Zealand. Metode ekstraksi menggunakan maserasi, metode fraksinasi menggunakan partisi cair-cair dengan corong pisah dan metode perlakuan pada hewan uji yang digunakan adalah pengolesan fraksi n-heksan, fraksi metanol, dan ekstrak daun talas pada luka terbuka kelinci New Zealand dengan kontrol positif dan kontrol negatif. Data yang didapatkan adalah berupa nilai penyempitan luka terbuka pada punggung kelinci. Hasil data dianalisis dengan ANOVA(Analysis Of Variant) yang dilanjutkan dengan uji LSD (Least Significant Different) untuk melihat perlakuan mana yang memberikan efek yang berbeda ditiap kelompok. Hasil penelitian atau pengujian selama 14 hari menunjukkan bahwa ekstrak, fraksi metanol, dan fraksi n-heksan mempunyai aktivitas penyembuhan luka pada punggung kelinci dengan data statistik yang menunjukkan adanya perbedaan signifikan antar kelompok perlakuan. Ekstrak mempunyai aktivitas daya sembuh luka yang paling optimal dibandingkan dengan kelompok perlakuan lain.Kata kunci : daun talas; ekstrak; fraksi; luka

Study of stiff differential equations of mathematical description of izomerization of pentane-hexane cut process

This paper provides a brief overview of the existing definitions of a stiff system of differential equations. Further, on the example of the accepted scheme of chemical transformations of the catalytic isomerization process of the pentane-hexane fraction, the stiffness of the system of differential equations was studied. In the course of the work, a method for studying the direct kinetic problem for stiffness is presented. In the Matlab software, the results of solving a system of differential equations by five methods (solvers) were compared. The given method can be tried for solving other problems of chemical kinetics.

Effect of Methanol Extract, Methanol Fraction, and N-Hexane Fraction of Parijoto Fruit (Medinilla speciosa) on Seminal Parameters and Testicular Weight of Male Sprague Dawley Rats

Infertility occurs in 2 million couples or 17% of couples who are married more than 2 years but are not pregnant or have signs of pregnancy. In couples who do not have children, 50% of male infertility factors are caused by abnormalities in semen. Empirically, parijoto fruits (Medinilla speciosa Blume) are used by the people of Kudus, Central Java, Indonesia to increase fertility. The purpose of this research was to find out the effect of methanol extract, methanol fraction, and n-hexane fraction of parijoto fruit (Medinilla speciosa) on seminal parameters and testicular weight of male Sprague Dawley rats. This research used 36 two-month-old Sprague Dawley rats with 200-300 gram of body weight which were divided into 6 groups. Group 1 (normal); groups 2, 3, and 4, used parijoto fruit methanol extract at 100mg/kgBW, 250mg/kgBW, and 500mg/kgBW doses respectively; groups 5 and 6 used methanol fraction and n-hexane fraction of parijoto fruit at 500 mg/kgBW dose respectively for 14 days. Rats were dissected and had examinations on sperm motility, morphology, abd testicular weight. The data were analyzed using Kruskal Wallis and Mann Whitney. The results found that groups 1, 2, 3, 4, 5 and 6 had mean spermatozoa motility (%) that were 56.5±2.43, 72.5±6.89, 77.6±12.99, 83.3±7.53, 84.7±3.98, and 74.2±11.58, mean spermatozoa morphology (%) that were 95.5±3.67, 95±2.76, 92.6±5.13, 96.5±3.27, 94.3±4.37, and 93.2±6.11, and mean testicular weight (gram) that were 1.08±0.10, 0.90±0.14, 0.98±0.10, 1.18±0.21, 1.28±0.43, and 1.02±0.13. There were significant differences between the normal group and all treatments (P <0.05) on spermatozoa motility. There was no significant difference on spermatozoa morphology and testicular weight. Based on the results, it can be concluded that methanol extract, methanol fraction, and n-hexane fraction of parijoto fruit can increase spermatozoa motility of male Sprague Dawley rats.

Effect of Enhydra fluctuans on Kidney Function in Alloxan-induced Diabetic Rats

AIM: The aim of this study was to explore the effect of n-hexane fraction of Enhydra fluctuans aerial on kidney function in alloxan induced diabetic rats. METHODS: Five groups of diabetic Wistar rats were studied: Group 1 was given 0.5% Na-CMC (G0), group 2 was given glibenclamide 0.45 mg/kg (G1), groups 3, 4, and 5 were given a dose of n-hexane fraction 57.03, 114.06, and 171.09 mg/kg respectively. The experiment was completed in 21 days. Blood glucose was estimated on day 0 and day 21 of treatment. Histology of kidney, creatinine, and blood urea nitrogen (BUN) was examined. ANOVA was used to evaluate quantitative data, which was then followed by Duncan's new multiple range test (p < 0.05). RESULTS: Our results demonstrate that n-hexane fraction dosages of 57.03 mg/kg and 114,06 mg/kg significantly improved blood glucose profile, BUN, and creatinine in diabetic rats. Moreover, the dosage of 57.03 mg/kg is effective to counteract necrosis and fibrosis of kidney cells. CONCLUSION: Our findings revealed that the administration of the n-hexane fraction of E. fluctuans aerial improved the kidney function of diabetic rats, especially at the dosage of 57.03 mg/kg. Therefore, E. fluctuans can be relied upon to be a drug to prevent the development of diabetes mellitus and diabetic nephropathy.