039180   GHAURI A O, AHMAD S, REHMAN T, BILAL M, ARSAD M A, ABBASI W M (Jinnah Women Univ, Pakistan, Email: aymenowais04@gmail.com) : Hypoglycemic and hypolipidemic effects of a poly herbal formulation in streptozotocin induced diabetic rats. Indian J Tradit Know 2020, 19, 137-44.

Diabetes mellitus is a chronic disease with several comorbidities. Currently used synthetic medicines have various side effects. Herbal medicines are being widely used for management of diabetes. The present study was conducted to evaluate the effect of poly herbal formulation (PHF) (mixture of Citrullus colocynthis, Syzygium cumini and Holarrhena antidysenterica) in diabetic rats. We evaluated enzyme inhibition activity of PHF by ELISA and anti-hyperglycemic action was assessed in streptozotocin induced diabetic rats. Animals were divided into 5 groups, i.e., blank, negative control, standard control and PHF 250 and 500 mg/kg dosage respectively. Blood glucose level assessed through one touch method blood glucose monitoring system. The statistical analysis was performed by using one-way ANOVA p value of ≤0.05 was considered significant. Results of study indicated that PHF sufficiently inhibited DPPH at 0.5 mg/mL and serial dilutions with IC₅₀ of 197 μg/mL while α- glucosidase inhibition assay showed marked inhibition of PHF at 0.5 mg/mL and serial dilutions with IC₅₀ of 235 μg/mL. Oral intake of PHF reduced blood glucose level, serum triglyceride and cholesterol levels in wistar albino rats in dose dependent manner. Considering the multidirectional effects of PHF, it is concluded that PHF may be a therapeutic option in management of diabetes.

3 illus, 4 tables, 46 ref

039177   KUMARI N, MENGHANI E, MITHAL R (Microbiology Dep, JECRC Univ, Jaipur- 303 905, Email: nalineek9@gmail.com) : GCMS analysis & assessment of antimicrobial potential of rhizospheric actinomycetes of AIA3 isolate. Indian J Tradit Know 2020, 19, 111-9.

Plants have been used for medicine to support human health in many regions in the world by researchers since ancient times. Plants and soil organisms have been found to have very high therapeutic potential as they produce many natural products. Evolving drug resistance towards nearly all anti-infection drugs, lead to the fast development of new drugs. Many natural products or secondary metabolites have been used for animal and human health. Recently, many new secondary metabolites from actinomycetes have been isolated and reported as important compounds with different activities like anti-microbial, anti-oxidant, anti-inflammatory, anti-androgenic and anticancer agents, etc. In this study isolation of actinomycetes was carried out on actinomycetes isolation agar media (AIA). Characterization and biochemical tests were performed and followed by fermentation and solvent extraction by four solvents for example- Benzene, pet ether, ethyl acetate, chloroform. GCMS was performed for identification of compounds present in culture broth. Major compounds present were Octanal,Pyrrolo[1,2-a]pyrazine-1,4-dione,hexahydro-3-(2-methylpropyl), Dibutyl phthalate, N-hexadecanoic acid, 1-nonadecene, Heptadecane, Octadecanoic acid, 3,7,11,15-tetramethyl-2-hexadecene, Dihydroergotamine, Hexadecanoic acid, 2-hydroxy-1-(hydroxymethyl) ethyl ester, Octadecanoic acid, 2,3-dihydroxypropyl ester, 13-docosenamide, and 4-tert-butylcalix[4]arene. Crude obtained was checked for their antimicrobial activity and inhibition zones (IZ) were noted on Mullar Hinton agar (MHA) media against indicator organisms like Staphylococcus aureus (MTCC-3160) (IZ=Ben-18 mm, E.A-25 mm), Pseudomonas aeruginosa (MTCC 1688) (IZ=Ben-11 mm, Chl-14 mm, E.A-24 mm), Klebsiella pneumonia (MTCC-432) (IZ=Ben-19 mm, Chl-20 mm, E.A-34 mm), Proteus vulgaris (MTCC-7306) (IZ=Benzene-10 mm, E.A-30 mm), Bacillus subtilis (MTCC-441). Identification of compounds was carried out by NIST 14 library.

2 illus, 4 tables, 32 ref

039158   KUMAR S D, SOJIN K, SANTHANAM P, DHANALAKSHMI B, LATHA S, KIM M-K (Daegu R & D Fusion Center, Daegu- 427 13, South Korea, Email: dinesk@bdu.ac.in) : Reciprocal response of nitrogen for enhancing growth and proximate compositions of marine microalga Tetraselmis sp. under low saline conditions. Indian J Geo-Mar Sci 2020, 49(2), 326-32.

The present investigation was aimed to study reciprocal response of nitrogen to enhance the cell multiplication and proximate balance from marine microalgae Tetraselmis sp. under low saline conditions. The strain has been isolated from Yellow Sea, South Korea and the algae were cultured in the different nitrogen concentrations (0N, 1N & 2N) under low saline conditions (15, 20 and 25 psu).The results revealed that the Tetraselmis sp. have significantly higher (P < 0.001) optical density, cell density, biomass concentration and total lipid in moderate nitrogen concentration (1N) under 25 psu of salinity condition. However, the total protein and carbohydrate were significantly higher (P < 0.001) in the high nitrogen concentration (2N) under 20 psu of salinity. The present study reveals that the combined effects made a huge impact on growth and biochemical variation in the microalgae.

4 illus, 1 table, 32 ref

039131   KIM M J, MOON J Y, KIM H S (Biological Science Dep, Ajou Univ, Korea, Email: hsunkim@ajou.ac.kr) : Phosphorylation of α-syntrophin is responsible for its subcellular localization and interaction with dystrophin in muscle cells. Indian J Biochem Biophys 2020, 57, 79-85.

Syntrophin is a well-known adaptor protein that links intracellular proteins with the dystrophin-glycoprotein complex (DGC) at the sarcolemma. However, little is known about the underlying mechanism that regulates the intracellular localization of α-syntrophin and its interaction with dystrophin. In this study, we demonstrate that α-syntrophin phosphorylation determines its intracellular localization and interaction with dystrophin in muscle cells. α-Syntrophin, a predominant isoform in skeletal muscles, directly interacts with ion channels, enzymes, receptors, and DGC proteins. Despite α-syntrophin being a potential signaling molecule, most studies focus on its function as a dystrophin-associated protein. However, we previously reported that α-syntrophin has a variety of DGC-independent functions to modulate cell migration, differentiation, survival, and protein stability. According to the results of the in vitro phosphorylation assays using subcellular fractions, the phosphorylated α-syntrophin accumulated only at the plasma membrane, and this event occurred regardless of dystrophin expression. However, the α-syntrophin interacting with dystrophin at the membrane was not in a phosphorylated state. We also identified that protein kinase C (PKC) was involved in the phosphorylation of α-syntrophin, which restricted α-syntrophin to interact with dystrophin. In conclusion, we demonstrate that the phosphorylation of α-syntrophin by PKC regulates its intracellular localization and interaction with dystrophin.

6 illus, 1 tables, 30 ref

039130   BALKHAIR A, GHAFARI A A, TASHKANDI H, ALTURKI S, DOGHAITHER H A, ABUSANAD A, ALKHAYYAT S, YEZERSKI A, ZEENELDIN A (Cancer and Mutagenesis Unit, Egypt, Email: abalghafari@kau.edu.sa) : Contribution of ABCG2 gene polymorphisms (G34A and C376T) in the prognosis of colorectal cancer. Indian J Biochem Biophys 2020, 57, 73-8.

This study aims to assess the association of two single nucleotide polymorphisms (SNPs), G34A and C376T, in the ABCG2 gene with the risk of developing CRC. To the best of our knowledge, this is the first study that determined the role of genetic variations in the ABCG2 gene with the risk of CRC in Saudi Arabia. The gDNA was extracted from the blood of 58 CRC patients and 48 healthy subjects. The DNA sequencing was used to determine the distribution of genotypes. The results showed that CRC patients carried a heterozygous (GA) genotype for SNP G34A had a low risk of developing CRC (odds ratio=0.015, 95 % CI [0.00–0.12]; risk ratio=0.35, 95 % CI [0.25–0.12], P <0.0001). On the other hand, patients that carried a heterozygous (CT) genotype for SNP C376T had a high risk of developing CRC (odds ratio=13.83, 95 % CI [4.31–44.38]; risk ratio=4.88, 95 % CI [1.95–12.24], P <0.0001). In conclusion, the results indicated that a heterozygous (GA) genotype in SNP G34A may decrease the risk of CRC development, whereas, the heterozygous (CT) genotype in SNP C376T may increase the risk of CRC. The results may suggest a protective role of ABCG2 SNP G34A against CRC and a deleterious effect of ABCG2 SNP C376T for increasing the risk of CRC.

3 tables, 27 ref

039129   OVEY I S, GULER Y (Physiology Dep, Alanya Alaaddin Keykubat Univ, Turkey, Email: suat.ovey@alanya.edu.tr) : Apoptotic efficiency of capecitabine and 5-fluorouracil on human cancer cells through TRPV1 channels. Indian J Biochem Biophys 2020, 57, 64-72.

The expression of transient receptor potential protein channels increases intensively colon and breast cancer cells. We aimed to reveal the role of 5-fluorouracil (5FU) and capecitabine along with Transient Receptor Potential Vanilloid 1 (TRPV1) channels in breast and colon cancer cells. Breast (MCF-7) and colon (Caco-2) cells were cultured and the study was planned as 7 main groups. Cells in the group were incubated with 5FU and capecitabine for 24 h and then incubated with TRPV1 channel antagonist capsazepine and stimulator capsaicin. The effects of medicines were investigated on molecular pathways of apoptosis. It was concluded that the administration of TRPV1 channel stimulator capsaicin in both cancer cells significantly increased the degree of intracellular Ca²⁺ levels and apoptosis levels compared to the control group whereas, the use of TRPV1 channel inhibitor capsazepine, significantly decreased the degree of apoptosis levels. The apoptotic effects of 5FU and capecitabine on both colon and breast cancer cells are directly related to TRPV1 channels and TRPV1 channels play an important role in the apoptosis. The apoptotic cell lines activity of capecitabine was higher in breast cancer cells, while that of 5FU was more pronounced in colon cancer cells.

3 illus, 2 tables, 34 ref

039128   GANESHPURKAR A, SALUJA A (AR Coll of Pharmacy, Vallabh Vidyanagar- 388 120, Email: akspharmacy@yahoo.com) : Immunomodulatory effect of rutin, catechin, and hesperidin on macrophage function. Indian J Biochem Biophys 2020, 57, 58-63.

The inflammation was observed by a sequence of comprehensive, correlated prevalence which causes enrollment of macrophages along with by removal of foreign particles and instigation of tissue repair. Though, massive release of chemokine and cytokines from macrophages can cause the progression of auto-inflammation and auto-immune diseases. The control overproduction of these mediators without damaging macrophages seems to be a novel strategy to control inflammation. Plants have been an integral part of the health-care system since ancient times. Many modern-day pharmaceuticals find their origin from phytochemicals. Rutin, catechin, and hesperidin are well-recognized for analgesic, anti-inflammatory and antiarthritic effects. Scientific reports rationalize their protective effect on humoral and cell-mediated immunity. The present study is focused to determine the immunomodulatory effect of rutin, catechin, and hesperidin on macrophage function. Rat peritoneal macrophages were harvested and cultured in the presence of lipopolysaccharides (LPS) and hesperidin. MTT assay was performed to determine macrophage viability. The levels of TNF-α, IL-1β, IL-6 were determined by ELISA kits whereas NO levels were determined by Griess method. The results of the present study revealed a decrease in levels of TNF-α, IL-1β, IL-6 and NO in LPS treated groups. In conclusion, LPS activated the macrophage by promoting the production of immune mediators, whereas rutin, catechin, and hesperidin treatment showed anti-inflammatory activities by suppressing the cytokine production in rat macrophages.

1 illus, 43 ref

039127   KANNAPAN S G P, RAGHUNATH G, SIVANESAN S, VIJAYARAGHAVAN R, SWAMINATHAN M (Anatomy Dep, Saveetha Institute of Medical and Technical Sciences, Chennai- 602 105, Email: senthilganesh52@gmail.com) : Inhibition of oxidative stress, inflammation and apoptosis by Terminalia arjuna against acetaminophen-induced hepatotoxicity in wistar albino rats. Indian J Biochem Biophys 2020, 57, 51-7.

Overuse of therapeutic drugs such as acetaminophen often affects liver, and may lead to inflammatory mediated liver cell death. Here, we studied the effect of Terminalia arjuna (TA) bark against acetaminophen (APAP) induced liver cell death/injury by testing the antioxidant levels, oxidative stress, and inflammation and apoptosis markers. Wistar albino male rats weighing 180-280 mg/kg were made into 5 groups of 6 animals each and were treated as follows: Gr. I, control; Gr. II, acetaminophen (APAP); GR. III, N-acetylcysteine (NAC); Gr. IV & V, Terminalia arjuna (TA) 250 and mg/kg. The antioxidant glutathione (GSH), lipid peroxidation (MDA), interleukin 1β (IL-1β) levels, caspase-9 levels, and Protein kinase B (P-AKT) gene expression levels were assessed. The rGr. V animals pre-treated with Terminalia arjuna high dose bark showed increased glutathione (GSH) levels, but decreased malondialdehyde (MDA) levels; inhibited IL-1β and caspase-9 levels; and also elevated gene expression level of P-AKT to regulate the cell signaling pathway. Apparently, the results demonstrated that a high dose of TA 500 mg/kg ameliorated acetaminophen-induced hepatotoxicity.

7 illus, 42 ref

039125   OTAIBI W A A, ALKHATIB M H, WALI A N (Regenerative Medicine Unit, Saudi Arabia, Email: mhalkhatib@kau.edu.sa) : Protective role of nanoemulsion containing Roman chamomile oil against mitomycin C-induced toxicity in Ehrlich ascites carcinoma bearing mice. Indian J Biochem Biophys 2020, 57, 33-44.

Mitomycin-C (MC), an anticancer agent, induces oxidative stress in normal tissues causing severe toxicity. The present study aimed to evaluate whether the side effects of MC may be diminished by the incorporation of MC into nanoemulsion containing Roman chamomile oil (RCM-NE). Fifty mice were equally divided into five groups. The first and second groups were the control and the untreated Ehrlich ascites carcinoma bearing mice (EAC), respectively, while the other three groups were EAC-bearing mice treated once intraperitoneally with a dose of 200 μL of RCM-NE, 1 mg/kg⁻¹ MC/200 μL normal saline, and 1 mg/kg⁻¹ MC/200 μL RCM-NE. The protective effect of the RCM-NE was examined by measuring the alterations in complete blood count, organs weight indices, and levels of serum biochemical parameters. The oxidative stress was assessed by measuring lipid peroxidation and enzyme activities of catalase, superoxide dismutase, glutathione reductase, and thioredoxin reductase-1. MC-RCM-NE has significantly reduced the toxicity of MC on the white blood cells and platelets. It improved the renal and cardiac biomarker levels and the antioxidants of the kidney, heart, and thymus. The protective effect of the RCM-NE on the blood and organs against MC toxicity was associated with the decreasing of oxidative stress and maintaining the activity of the antioxidants.

7 illus, 1 table, 33 ref

039124   SARKAR S, PAL A, CHOUNI A, PAUL S (Botany Dep, Calcutta Univ, Kolkata-712 136, Email: spaul_1971@yahoo.com) : A novel compound β-sitosterol-3-O-β-D-glucoside isolated from Azadirachta indica effectively induces apoptosis in leukemic cells by targeting G0/G1 populations. Indian J Biochem Biophys 2020, 57, 27-32.

Azadirachta indica, popularly known as  Neem, is an important plant in the Ayurveda system of medicine. It is known to prevent about at least 40 types of diseases in home-practice. A previous report showed that the methanolic extract of this plant can effectively control proliferation of leukemia cells. Here, we explored the eEthanolic extract dried neem leaves for a new molecule with potent anti-leukemic property.Four biomolecules were isolated viz. rutin, isoquercetin, quercetrin and β-sitosterol-3-O-β-D-glucoside from the ethanolic extract by repeated column chromatography and HPLC.Quercetrin structures of the isolated molecules were confirmed by Mass, ¹HNMR, ¹³C NMR spectra analysis. MTT assay revealed that β-sitosterol-3-O-β-D-glucosideeffectively reduced the proliferation of MOLT 4 leukemic cells in a dose and time dependent manner. DAPI staining with confocal microscopy indicated that β-sitosterol-3-O-β-D-glucoside efficiently induced nuclear DNA fragmentation in MOLT 4 cells. Finally, flow cytometry after PI staining showed that the compound has potential to check the cell cycle progression at sub G1 phase. With the available results, it can be concluded that β-sitosterol-3-O-β-D-glucoside has potential to be good therapeutic drug in leukemia treatment.

4 illus, 24 ref

039123   SHARMA S, MISHRA V, SRIVASTAVA N (Jiwaji Univ, Gwalior- 474 011, Email: nalinis21@yahoo.com) : Protective effect of Trigonella foenum-graecum and Cinnamomum zeylanicum against diabetes induced oxidative DNA damage in rats. Indian J Biochem Biophys 2020, 57, 15-26.

Diabetes mellitus has become a global epidemic as its prevalence is steadily increasing everywhere at an alarming rate, most markedly in the India. Traditional Indian system of medicine based on herbal therapies is being practiced since ancient times and has been proved very effective against number of diseases. Trigonella foenum-graecum (Tfg) and Cinnamomum zeylanicum (Cz), the two spices, possess multiple medicinal properties. Hence present study is aimed to evaluate the protective effects of these plants on the oxidative DNA damage in diabetic rats. The presence of DNA base oxidation was investigated by modified comet assay which uses bacterial repair enzymes, formamidopyrimidine glycosylase and Endonuclease III which can measure oxidatively damaged purines and pyrimidines, respectively. Diabetes induced alteration in total antioxidant capacity (TAC), generation of superoxide anion and the levels of GSH, GSSG, NADH, NAD⁺, NADPH and NADP⁺ were monitored. Diabetes was induced by intravenous injection of alloxan, followed by oral administration of aqueous suspension of Tfg seeds and bark of Cz. Treatment of Tfg and Cz significantly restored the altered level of superoxide anion, TAC, ratios of GSH/GSSG, NAD(P)H/NAD(P)⁺ and NADH/NAD⁺ in the diabetic group. Furthermore, both the treatment effectively decreased the level of oxidatively damaged purine and pyrimidine bases.

2 illus, 4 tables, 49 ref

039119   KUMAR S S, SUDARSHAN M, CHAKRABORTY A (Physics Dep, Kanchi Mamunivar Centre for Post Graduate Studies (Autonomous), Puducherry- 605 008, Email: santhosh.physics@gmail.com) : Trace elements analysis of Biophytum sensitivum (L.) DC. by EDXRF spectroscopy and its therapeutic role in Diabetes mellitus. Indian J Biochem Biophys 2020, 57, 8-14.

Minerals and trace elements, as structural components of tissues, play vital role in cellular and basal metabolism. The Little Tree Plant, Biophytum sensitivum (L.) DC., locally called Lajalu, is a well-known medicinal herb, traditionally considered as one of the 10 flowers used since ancient time for healthy life. In Ayurveda, it is used to treat various ailments viz., asthma, insomnia, convulsions, cramps, stomach ache, chest complaints, inflammation, tumours and skin disorders. In this study, we analyzed the elemental contents of its leaves and roots using EDXRF spectroscopy, and tried to find possible correlation with its nutritional and curative effects. The elements Ca, K, Si, Cl, P, and S were found in the highest concentration in leaf, in the descending order, but in the root, K took the leading position. The results have been discussed with reference to the established role of these elements in physiology and pathology of human life with special focus on its role in bone building, controlling the sugar level and wound healing capacity of patients with diabetes mellitus.

2 illus, 1 tables, 46 ref

039114   NEGRETA C P, ACOSTA M A F, MULIA J, ROSAS L A M, GONZALEZ D O (Autonomous Univ, Mexico, Email: dog@uaemex.mx) : Structural variations induced by temperature changes in rotavirus VP6 protein immersed in an electric field and their effects on epitopes of the region 300-396. J Nucl Phys Mat Sci Rad A 2020, 7(2), 189–94.

Rotavirus diarrhea is an infectious intestinal disease that causes about 215 thousand deaths annually in infants under five years old. This virus is formed by three layers of concentric proteins that envelop its genome, from which VP6 structural protein is the most conserved among rotavirus serotypes and an excellent vaccine candidate. Recent studies have shown that structural proteins are susceptible to losing their biological function when their conformation is modified by moderate temperature increments, and in the case of VP6, its antigen efficiency decreases. We performed an in silicoanalysis to identify the structural variations in the epitopes 301-315, 357-366, and 376-384 of the rotavirus VP6 protein-in a hydrated medium- when the temperature is increased from 310 K to 322 K. In the latter state, we applied an electric field equivalent to a low energy laser pulse and calculated the fluctuations per amino acid residue. We identified that the region 301-315 has greater flexibility and density of negative electrical charge; nevertheless, at 322 K it experiences a sudden change of secondary structure that could decrease its efficiency as an antigenic determinant. The applied electric field induces electrical neutrality in the region 357-366, whereas in 376-384 inverts the charge, implying that temperature changes in the range 310 K-322 K are a factor that promotes thermoelectric effects in the VP6 protein epitopes in the region 300-396.

4 illus,1 tables, 27 ref

039112   LI Q, DOU W, QI J, QIN X, CHEN S, HE Y (Southwest Univ, China, Email: chenshanchun@cric.cn) : Genomewide analysis of the CIII peroxidase family in sweet orange (Citrus sinensis) and expression profiles induced by Xanthomonas citri subsp. citri and hormones. J Genet 2020, 99, 1-10.

Class III peroxidase (CIII prx) is a plant-specific multigene family that regulates the physiological and stress responses. This research aimed to exhaustively annotate and analyse the CIII prx family in sweet orange and to explore the regulated expression profiles by Xanthomonas citri subsp. citri (Xcc) and plant hormones. We further assessed the relationship between CIII prxs and citrus bacterial canker. The phylogeny, gene structure, conserved motifs, gene duplications and microsynteny of the CIII prx family were analysed. Expression profiles of specific CsPrxs induced by Xanthomonas citri subsp. citri and plant hormones were detected by quantitative reverse transcription-polymerase chain reaction. Subcellular localization was analysed through transient expression assessments. A total of 72 CIII prx members were identified from the genomes of sweet orange. In all chromosomes of sweet orange, the CsPrxs could be detected except in chromosome 8. In addition, three segmental duplications, four tandem duplications and 11 whole-genome duplications occurred among the CsPrxs, contributing to the family size expansion. From the K_a/K_s ratios, 15 of 18 duplicated CsPrxs pairs have experienced purifying selection process. A total of 15 conserved motifs were detected in CsPrxs, four of which were detected in all complete CsPrxs. A total of 12 expressed genes were identified from the EST database. The expression trends of 12 CsPrxs were differently expressed at different stages of infection by Xcc, five of which were potential candidate genes involved in Xcc resistance. These genes could be induced by salicylic acid and methyl jasmonate, and were extracellular proteins. These results further support our understanding of CIII prxs in citrus, particularly in citrus bacterial canker studies.

8 illus, 1 tables, 33 ref

039105   LIU H, LU Y, LAN B, XU J (Beijing Forestry Univ, China, Email: jcxu282@sina.com) : Codon usage by chloroplast gene is bias in Hemiptelea davidii. J Genet 2020, 99, 1-8.

The base composition of the chloroplast genes is of great interest because they play a highly significant role in the evolutionary development of the plants. Evaluation of the 48 chloroplast protein-coding genes of Hemiptelea davidii showed that the average GC content was about 37.32 %, while at the third codon base position alone the average GC content was only 27.80 %. The 48 genes were classified into five groups based on the gene function and each group displayed specific codon characteristics. Based on the relative synonymous codon usage analysis, a total of 30 high-frequency codons and 11 optimal codons were identified, most of them ended with A or T. Neutrality plot, ENC-plot and PR2-plot analyses showed that the codon usage bias of the chloroplast genes of H. davidii was greatly influenced by natural selection pressures. Meanwhile, the frequency of codon usage of chloroplast genes among different plant species displayed similarities, with some synonymous codons were preferred to be used in H. davidii. In this study, the codon usage pattern of the chloroplast proteincoding genes of H. davidii provides us with a better understanding of the expression of chloroplast genes, and may advice the future molecular breeding programmes.

4 illus, 6 tables, 42 ref

039104   GARCIA M E R, VINAGRE F J C, CANALES E A, ARAGON A M D, SACHEZ L H, FORNES F R, RODRIGUEZ P C, CONEJO M M, HERNANDEZ E M, AZORIN F M (Centro de Investigacio´n Biome´dica en Red de Enfermedades Raras, Spain, Email: fmartinez@h12o.es) : A novel de novo mutation in the PURA gene associated with a new clinical finding: Large brainstem. J Genet 2020, 99, 1-7.

We report the case of a Caucasian Spanish boy, who showed profound neonatal hypotonia, feeding difficulties, apnea, severe developmental delay, epilepsy, bilateral convergent strabismus, poor verbal language development and a large brainstem. Whole-exome sequence uncovered a novel de novo mutation in the purine-rich element binding protein A gene (PURA; NM_005859.4:c.72del:p.(- Gly25AlafsTer53)) that encodes the transcriptional activator protein Pur-alpha (PURA). Mutations in this gene have been identified in patients with PURA syndrome, a rare disorder characterized by an early hypotonia, developmental delay, severe intellectual disability with or without epilepsy, and disability in expressive language development. Although, up to 75 cases have been identified worldwide, to the best of our knowledge, this is the first patient described with a brainstem larger than normal. In conclusion, our data expand both genetic and phenotypic spectrum associated with PURA gene mutations.

2 illus, 1 table, 15 ref

039103   ALINY Z, AKBARI M T, DEILAMANI F K (Tehran Medical Genetics Laboratory, Iran, Email: akbarimt@gmail.com.) : Assessment of FMR1 triplet repeats in patients affected with mental retardation, fragile X syndrome and primary ovarian insufficiency. J Genet 2020, 99, 1-6.

The CGG repeats in the FMR1 gene expand in patients with fragile X syndrome, fragile X-associated tremour/ataxia syndrome and fragile X-associated primary ovarian failure. In this study, the CGG repeats in the FMR1 gene were studied in 449 males and 207 females using traditional polymerase chain reaction and triplet repeat primed PCR methods, also 18 CVS samples (six males and 12 females) were tested for prenatal diagnosis. Further, methylation sensitive multiplexed ligation dependent probe amplification was performed on some samples to confirm the results. Regarding the male patients, 1.1 % and 9.7 % had premutation (PM) and full mutation (FM) alleles, respectively. Also three (0.66 %) male patients were mosaic for PM and FM alleles. Among females, 1.9 % were GZ carriers and 5.8 % were PM carriers. Prenatal diagnosis resulted in detection of two PM and one FM males as well as one FM carrier female. Our results were in concordance with the previously published results.

1 illus, 2 tables, 24 ref

039010   MORSHEDY S A, BASYONY M, ABDELMODATHER A, ZAHRAN S A, HASSAN M A (Fish and Animal Production Dep, Alexandria Univ, Egypt) : Effect of oral administration of wheat (Triticum aestivum L.) germ, Eruca sativa Mill. seed oils and their mixture on semen evaluation of V-line rabbit bucks. Asian J Anim Sci 2020, 14(2), 44-53.

Different natural feed additives have been used to maintain semen quality characteristics of rabbit bucks at heat stress. This study was carried out to investigate the effect of administration of vegetable oils on the semen quality of male rabbits. Twenty adult V-Line rabbit bucks 6-7 months old, with average initial body weight (BW) 2792.40±66.39 kg were randomly distributed into four experimental groups (n = 5). The control group administered with 1.0 mL of distilled water, the second group received rocket seed oil (0.3 mL kg^-1 b.wt., Rocket Seed Oil (RSO)), the third group received wheat germ oil (0.3 mL kg^-1 b.wt., Wheat Germ Oil (WGO)) and the fourth group received 0.15 mL of RSO and 0.15 mL of WGO kg^-1- b.wt., mixture. The rabbits  were  given  oils orally once  daily  for  8  weeks  under  summer  conditions  (27.50EC  and  76 %  relative  humidity). Feed  intake  of  rabbits  decreased (p = 0.0001) with orally received Rocket Seed Oil (RSO) and didn’t differ with Wheat Germ Oil (WGO) or their mixture compared to the control group. Body weight change was tended to increase in rabbits received RSO compared to the control group. The treatments with oils significantly improved semen quality compared to the control group. The oral administration of RSO, WGO and their mixture improved semen quality characteristics under summer conditions induced heat stress.

2 illus, 3 tables, 68 ref

039007   MAHMOUD U T, EL-SAYED M S A, KHALIL N S A (Animal and Poultry Behavior and Management Dep, Assiut Univ, Assiut- 71526, Egypt) : Litter number modulates some behavioral and physiological aspects in Egyptian native breed ewes (Ovis aries). Asian J Anim Sci 2020, 14(1), 25-32.

Lactation is considered as one of the most stressful conditions along the entire life of  female mammals. From the welfare point of view, nursing several lambs represents emerging health challenge both on the levels of dam and offspring. Therefore, this study was designed to explore the effects of litter number on Egyptian native breed sheep welfare by evaluating some physiological and behavioral parameters in ewes from 0D-30D after giving birth. Twelve multiparous ewes were identified into 2 groups based on their litter  number as follow: Single Lamb Ewe (SLE) group include 6 ewes, each  one  had  birth  for one lamb and Twin Lamb Ewe (TLE) group include 6 ewes, 5 ewes each one of them had birth for two lambs and one ewe had birth for 3 lambs). TLE group ewes had significantly higher total movement and maternal rejection behavioral activities and significantly lower resting and total ingestive behavioral activities than SLE group ewes. In addition, results showed that TLE group at days 0 and 30 postpartum had significantly higher levels of cortisol and prolactin (PRL) and lower calcium level than SLE group. At day 0, the plasma progesterone (P4) and glucose levels were significantly higher in TLE group than that of SLE group. Increased litter number may affect negatively on the behavioral and physiological conditions of  ewes as it put them under stress.
 

2 illus, 2 tables, 51 ref

039005   MPENDA F N, LYANTAGAYE S L, BUZA J (Global Health Dep, Nelson Mandela African Institution of Science and Technology, Arusha, Tanzania) : Association of chicken mx1 polymorphisms with susceptibility in chicken embryos challenged with virulent newcastle disease virus. Asian J Anim Sci 2020, 14(1), 12-21.

 Genetic selection of chicken genotypes that are less susceptible to Newcastle disease virus (NDV) is a promising  option.  The  present  study  aimed  to  investigate  polymorphism  of  chicken  Mx1 gene G2032A SNP in different chicken breeds in Tanzania and association with chicken embryos susceptibility to virulent NDV. A  total of 355 (87 Sasso, 129 Kuroiler and 139 local) 16 days old chicken embryos were infected with 0.1 mL minimum lethal dose (103/0.1 mL) of virulent NDV suspension. The time of death was recorded and selective genotyping was deployed where chicken embryos from high (15 %) and less (15 %) susceptible cohorts were genotyped by PCR-length polymorphism. Two forward and a reverse primers were used to generate 2 different sizes of PCR product of A and G alleles of chicken Mx1 gene G2032A SNP. As expected chicken embryos survival was highly variable within a breed. The frequency of allele A was higher than allele G in chicken breeds. Furthermore, using Pearson’s X2 test of independence, it was demonstrated that chicken Mx1 gene G2032A genotypes (AA, AG and GG) were associated (p<0.05) with chicken embryos susceptibility to virulent NDV infection. The frequency of genotype AA was higher in less susceptible chicken embryos, whereas, frequency of genotype GG and AG was higher in high susceptible chicken embryos.  Results from the present study provide valuable information on the potential role of Mx1 gene polymorphism for selection of chickens that are resistant to virulent NDV infection.
 

2 iilus, 4 tables, 36 ref

039004   NAJIB H, AL-HOMIDAN I, FATHI M M, AL-SUHIM A A (Animal and Fish Production Dep, King Faisal Univ, Hofuf- 31982, Saudi Arabia) : Black seeds (Nigella sativa) and ginger powder (Zingiber officinale) effect on growth performance and immune response of broiler chickens. Asian J Anim Sci 2020, 14(1), 1-8.

 Antibiotic has been banned for animal use since 2006. Some studies provided evidences that some medicinal plants have antimicrobial properties and can replace antibiotics. This study was carried out to investigate the effect of  adding 3 levels of black seeds and 3 levels of ginger root powder in broiler diets (starter and grower) on productive performance, immunity indicators and bacterial population of small intestine. A total of 700 day-old unsexed broiler chicks (Ross 308) were randomly allocated into 7 groups: A control group (corn-soybean basal diet with no supplement) and 6 dietary treatment groups receiving 10, 20 and 30 g kg^-1 of black seeds and 10, 20 and 30 g kg^-1 of ginger root powder. These chicks were distributed into 5 floor pens (replicates)/treatment (20 chicks each).This study showed that black seed may play a potential role as an antioxidant in the components of blood more than ginger root powder does. Both black seed and ginger powder have a significant inhibitory effect on pathological bacteria in broiler's gut. Black seeds can be used as a feed additive to improve live body weight, antioxidant profile and gut microbiota of broilers. The recommended dosage of BS in the diets of broilers is 20 g kg^-1. Although, ginger powder did not affect growth performance and immune response, it can be used for improving antioxidant status and microbiota profile in broilers.

6 tables, 45 ref

006603   KASANGANA P B, EID H M, NACHAR A, STEVANOVIC T, HADDAD P S (Wood Sciences Dep, Laval Univ, Quebec, Canada, Email: pierre-betu.kasangana.1@ulaval.ca) : Further isolation and identification of anti-diabetic principles from root bark of Myrianthus arboreus P. Beauv.: The ethyl acetate fraction contains bioactive phenolic compounds that improve liver cell glucose homeostasis. J Ethnopharmacol 2019, 245, 112167.

We recently reported that ethanol extract of Myrianthus arboreus P. Beauv. root bark demonstrated antidiabetic activity by modulating hepatocyte glucose homeostasis. This activity was associated significantly to the ethylacetate (EAc) fraction. The current study sought to identify the active compounds responsible of the antidiabetic effect of M. arboreus in the EAc fraction using bioassay-directed sub-fractionation. EAc fraction was sub-fractionated using Flash chromatography. Preparative HPLC was used to isolate the pure compounds. The structures of the isolated compounds were confirmed by analysis of NMR spectroscopic and mass spectrometric data. Hepatic (H4IIE, HepG2) cells were treated with maximum non-toxic concentrations of ethanol extract, its EAc fraction and isolated compounds thereof. Glucose-6-phosphatase (G6Pase) activity was measured using the glucose oxidase method. To measure glycogen synthase (GS) activity, radioactive assays were used. Phosphorylation of AMP-activated protein kinase (AMPK) and Glycogen Synthase Kinase-3 (GSK-3) were probed by Western blot. Six sub-fractions were obtained, and the antidiabetic activity was found in two sub-fractions (SFE1 and SFE2). For the first time, two known C-glycosylflavone regio-isomers, isoorientin (1) and orientin (2) were detected and isolated from M. arboreus plant, especially from SFE2 as well as protocatechuic acid, 3,4-dihydroxybenzaldehyde (4), and chlorogenic acid isolated from SFE1. The compounds 1, 2, 4 were determined to decrease the activity of G6Pase by increasing AMPK phosphorylation and to stimulate GS through GSK-3 phosphorylation. Isoorientin which is one of the main compounds of EAc fraction, expressed the strongest effect in all bioassays, similar to that of the EAc fraction. A significant and linear correlation was found between the phosphorylation of AMPK and the activity of G6Pase modulated by all samples (R² = 0.54; p < 0.05). Similar to G6Pase assay, a correlation was determined between the capacity of M. arboreus extracts/fractions/compounds to stimulate GS activity and to phosphorylate GSK-3 (R² = 0.57, p < 0.01). Results demonstrate that 1, 2, 4 are responsible, at least in part, for the antidiabetic activity of M. arboreus. These compounds can be used to ensure the quality and efficacy of M. arboreus antidiabetic preparations and standardize such preparations.

37 ref

006581   ESAKKIMUTHU S, NAGULKUMAR S, DARVIN S S, BUVANESVARAGURUNATHAN K, SATHYA T N, NAVANEETHAKRISHAN K R, KUMARAVEL T S, MURUGAN S S, SHIROTA O, BALAKRISHNA K ET.AL (Tokushima Bunri Univ, Kagawa- 769-2193, Email: oshirota@kph.bunri-u.ac.jp) : Antihyperlipidemic effect of iridoid glycoside deacetylasperulosidic acid isolated from the seeds of Spermacoce hispida L. - A traditional antiobesity herb. J Ethnopharmacol 2019, 245, 112170.

The interest on herbal health supplements for obesity is increasing globally. Our previous ethnobotanical survey in Tiruvallur district, Tamil Nadu, India indicated the use of Spermacoce hispida L. seeds for the treatment of obesity. This study was aimed to validate the traditional claim and to identify the antihyperlipidemic principle in the seeds of Spermacoce hispida using bioassay guided fractionation method. Bioassay monitored fractionation of the aqueous extract from Spermacoce hispida seeds was carried out using triton WR 1339 induced hyperlipidemic animals. It yielded deacetylasperulosidic acid (DAA) as the active ingredient. Pharmacokinetic properties of DAA were predicted using DataWarrior and SwissADME tools. In vitro antiobesity and antihyperlipidemic effects of DAA were evaluated in 3T3L1 preadipocytes and HepG2 cells, respectively. The chronic antihyperlipidemic efficacy of DAA was evaluated in high fat diet fed rats. DAA did not show any mutagenic and tumorigenic properties. It bound with PPARα with comparable ligand efficiency as fenofibrate. The treatment with DAA significantly lowered the proliferation of matured adipocytes, but not preadipocytes. The treatment of steatotic HepG2 cells with DAA significantly decreased the LDH leakage by 43.03% (P < 0.05) at 50 μM concentration. In triton WR 1339 induced hyperlipidemic animals, the treatment with 50 mg/kg dose significantly lowered the TC, TG and LDL-c levels by 40.27, 46.00 and 63.65% respectively. In HFD fed animals, the treatment at 10 mg/kg decreased BMI and AC/TC ratio without altering SRBG. It also improved serum lipid, transaminases and phosphatases levels of HFD fed animals. The treatment lowered adipocyte hypertrophy and steatosis of hepatocytes.This preliminary report supported the traditional use of Spermacoce hispida for the treatment of obesity. Further detailed investigations on the long term safety, efficacy and molecular mode of action of Spermacoce hispida and DAA will throw more light on their usefulness for the management of obesity. 

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006639   SABEL R, FRONZA A S, CARRENHO L Z B, MAES A, BARROS M L, POLLO L A E, BIAVATTI M W, HERDE K D, VANDENABEELE P, KREUGER M R O (Centro Univ, Avantis, Brazil, Email: lzbcarrenho@gmail.com) : Anti-inflammatory activity of the sesquiterpene lactone diacethylpiptocarphol in dextransulfate sodium-induced colitis in mice. J Ethnopharmacol 2019, 245, 112186.

Sesquiterpene lactones are organic compounds derived mainly from plants that exhibit anti-inflammatory and antitumor activities being one of the key mechanism of action of NF-kB pathway and synthesis of cytokines such as IL-1 and TNF- α. The overall objective of the present study was to evaluate the anti-inflammatory action of a sesquiterpene lactone diacethylpiptocarphol (DPC) from Vernonia scorpioides (Lam.) Pers. and parthenolide (PTH) in Balb-c mice with DSS-induced colitis. The anti-inflammatory effects of Intraperitonial administration of DPC (5 mg/kg/day) were evaluated in Balb/c mice with DSS-induced colitis, and further the body weight measurement, TNF-α and TGF-β level was determined. After intraperitoneal treatment for one week, DSS-induced colitis was significantly reduced in mice treated with either of both sesquiterpenes lactones, as witnessed by reduced cellular infiltration, tissue damage, TNF-α production, and enhanced production of TGF-β. Sesquiterpene lactone DPC, isolated from Vernonia scorpioides showed anti-inflammatory activity, in this experimental model of colitis the sesquiterpene lactones DPC and PTH exhibit equal anti-inflammatory activity.

28 ref

006610   LI P, LIU S, LIU Q, SHEN J, YANG R, JIANG B, HE C, XIAO P (Chinese Academy of Medical Sciences, Beijing- 100 193, Email: lee_p1214@163.com) : Screening of acetylcholinesterase inhibitors and characterizing of phytochemical constituents from Dichocarpum auriculatum (Franch.) W.T. Wang & P. K. Hsiao through UPLC-MS combined with an acetylcholinesterase inhibition assay in vitro. J Ethnopharmacol 2019, 245, 112185.

The genus Dichocarpum is endemic to East Asia, and many of them are traditionally used folk medicine in China. Dichocarpum auriculatum (Franch.) W. T. Wang et P. K. Hsiao has the effect of clearing away heat, removing toxicity, and relieving swelling in southwestern China. Intriguingly, its root and whole herb also used as remedy for the neurological disease epilepsy. However, there are not any scientific reports on the phytochemistry and pharmacological activities of D. auriculatum. Traditional and folk medicinal knowledge would be useful for finding new pharmaceutical resources. There are many evidences over the years reported that an interaction probably exists between epilepsy and Alzheimer's disease (AD). The aim of the study was to investigate the potential AChE inhibitors and the phytochemical profiles of the specie D. auriculatum. The AChE inhibitory activity of plant extracts of D. auriculatum and other 6 species from different regions of the genus Dichocarpum were evaluated in vitro assays and the UPLC-Q-TOF-MS technique was used to analyze the chemical constituents. Moreover, UPLC-ESI-MS/MS was used to determine the distribution of 12 standard compounds in samples. As a preferred source of potential acetylcholinesterase inhibitors of the genus Dichocarpum, D. auriculatum has been further investigated. The screening results show that the ability of root extracts from D. auriculatum (IC₅₀ = 0.15 mg·mL^-1) to inhibit AChE was better than other samples, it is consistent with traditional medicinal records. The phytochemical constituents of D. auriculatum was surveyed firstly by UPLC-Q-TOF-MS analysis, and 36 compounds, including 14 alkaloids, 16 flavonoids, 6 others, were identified tentatively. Further experiments showed that five compounds (columbamine, palmatine, dauricine, jatrorrhizine and berberine) from D. auriculatum were confirmed the potential inhibition of AChE activity in vitro (IC₅₀: 0.24–6.37 μM) and UPLC-ESI-MS/MS results showed that the content of most active compounds in roots was much higher than in aerial parts. Palmatine (IC₅₀ = 0.34 μM) and columbamine (IC₅₀ = 0.24 μM) showed prominent AChE inhibitory activity among the tested compounds. This is the first report about the evaluation of AChE inhibitory activity and phytochemical profiles of D. auriculatum, led to the identification of 36 compounds including alkaloids and flavonoids, and five alkaloids exhibited a significant AChE inhibitory activity and had the potential as AChE inhibitors. This study provided scientific experimental basis for the traditional efficacy of neurological disease of the plant.

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006625   OYEDEMI B O, KOTSIA E M, STAPLETON P D, GIBBONS S (Research of Pharmaceutical and Biological Chemistry Dep, UCL School of Pharmacy, London, UK, Email: simon.gibbons@ucl.ac.uk) : Capsaicin and gingerol analogues inhibit the growth of efflux-multidrug resistant bacteria and R-plasmids conjugal transfer. J Ethnopharmacol 2019, 245, 111871.

Capsicum and ginger are used widely in human diets and in folklore medicines. Chemically, gingerol is a relative of capsaicin and both classes of compounds are notable for their spiciness and characteristic pungent aroma. Previous studies have demonstrated that these compounds contain antimicrobial compounds with robust pharmacological importance. The present study evaluated the in vitro antibacterial activities of capsaicinoids and gingerols against a panel of clinical MRSA strains and their inhibitory effect on the conjugal transfer of R-plasmids harboured in Escherichia coli Crude methanol extract of C. annum was fractionated using solid phase extraction (SPE) and screened for R-plasmid transfer inhibition: TP114, PUB 307, PKM 101, R6K and R7K. The bio-guided assay led to the isolation of bioactive compounds with strong R-plasmid transfer inhibition. The compounds were identified using Nuclear Magnetic resonance (NMR) and Mass spectroscopy (MS). Capsaicin analogues nonivamide, 6-gingerol, 6-shogaol, capsaicin and dihydrocapsaicin were screened for antimicrobial activity against a panel of methicillin-resistant Staphylococcus aureus (MRSA) and Gram-negative bacteria strains using microdilution method while the plasmid transfer inhibition assay of the compounds was determined by broth mating method. The bioactive fraction Ca-11 showed good inhibition rates (8.57–25.52%) against three R-plasmids PUB307, PKM 101, TP114 followed by the crude extract of C. annum (8.59%) respectively leading to the bioassay-guided isolation of capsaicin and dihydrocapsaicin as the bioactive principles. The antiplasmid effect of pure capsaicin and dihydrocapsaicin were broad and within active ranges (5.03–31.76%) against the various antibiotic resistance-conferring plasmids including R6K, R7K. Capsaicin, 6-gingerol and 6-shogaol had good broad antibacterial activity with MIC values ranging from 8 to 256 mg/L against effluxing MRSA strains SA1199B (NorA), XU212 (TetK) and RN4220 (MsrA). While they exhibited moderate antibacterial activity (128–512 mg/L) against the Gram-negative bacteria. The effect of 6-gingerol, 6-shogaol and nonivamide on the plasmids were very active on PKM 101 (6.24–22.16%), PUB 307 (1.22–45.63%) and TP114 (0.1–7.19%) comparative to the positive control plumbagin (5.70–31.76%). These results are suggestive that the R-plasmids could possess substrate for capsaicinoids-like compounds and for their ability to inhibit the plasmid conjugation processes. Plant natural products possess the potential value of antibacterial and mechanistic antiplasmid activity as demonstrated by the compounds and should be evaluated in developing antimicrobial leads to novel mechanism against multidrug-resistant bacteria.

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006563   BARBOSA C C, RODRIGUES T C, ATAIDES C F S, SANTOS M L, GHEDINI P C, JUNIOR W D, ANDERSON M L, MAZARO-COSTA R (Pharmacology Dep, Federal de Goiás Univ, GO, Brazil, Email: mazaro.renata@gmail.com) : Protective effects of Hydrocotyle umbellata var. bonariensis Lam. (Araliaceae) on memory in sleep-impaired female mice. J Ethnopharmacol 2019, 245, 112183.

Hydrocotyle umbellata var.bonariensis Lam. (Hb), popularly known in Brazil as acariçoba and outside Brazil Hb by a number of names including marsh-pennywort, and many-flower, has traditionally been used in Ayurvedic medicine in the retardation of aging (Rasayana effect). The present study evaluated the effect of Hb treatment before and during paradoxical sleep deprivation (PSD) and sleep restriction (SR) on learning, memory, and acetylcholinesterase (AChE) brain activity. Adult Swiss nulliparous female mice were randomly distributed among the experimental groups. The treated groups received the aqueous solution of Hb leaves orally at concentrations of 500 and 1.000 mg/kg. PSD and SR were induced by the multiple platform method, in which the animals remained for 3-days in PSD or 15-days in SR for 22 h per day. The collection of the vaginal epithelium occurred daily to determine the estrous cycle. Body mass gain was determined. The animals were submitted to the passive avoidance test and were then euthanized for the collection of brain tissue and the determination of cerebral cholinesterase activity. The aqueous solution of Hb was associated with a significant reduction in cholinesterase activity at both doses in the SR model, and at the dose of 1.000 mg/kg in the PSD model. Regarding the learning and memory test, the PSD group treated with 1.000 mg/kg presented significant improvement, whereas in the SR experiment none of the treated-groups showed any improvement in learning and memory. In the analysis of SR/PSD interference and/or Hb treatment on the estrous cycle, it was possible to observe that the treatment acted as a protector in the SR group, maintaining a normal cycle. The analyses showed that Hb was safe to use during periods of SR or PSD, acting as an adaptogen for these situations, in addition to being able to reduce cholinesterase activity, which suggests its neuroprotective action. In relation to the estrous cycle, Hb can act as a protector in SR situations.

8 illus, 45 ref

006624   OMAR W F N W, GIRIBABU N, KARIM K, SALLEH N (Physiology Dep, Malaya Univ, Kuala Lumpur, Malaysia, Email: naguibsalleh@um.edu.my) : Marantodes pumilum (Blume) Kuntze (Kacip Fatimah) stimulates uterine contraction in rats in post-partum period. J Ethnopharmacol 2019, 245, 112175.

Marantodes pumilum (Blume) Kuntze has traditionally been used to firm the uterus after delivery, however scientific evidences behind this claim is still lacking. To demonstrate Marantodes pumilum leaves aqueous extract (MPE) has an effect on uterine contraction after delivery and to elucidate the molecular mechanisms involved. Day-1 post-delivery female rats were given MPE (100, 250 and 500 mg/kg/day) orally for seven consecutive days. A day after the last treatment (day-8), rats were sacrificed and uteri were harvested and subjected for ex-vivo contraction study using organ bath followed by protein expression and distribution study by Western blotting and immunohistochemistry techniques, respectively. The proteins of interest include calmodulin-CaM, myosin light chain kinase-MLCK, sarcoplasmic reticulum Ca²⁺-ATPase (SERCA), G-protein α and β (Gα and Gβ), inositol-triphosphate 3-kinase (IP3K), oxytocin receptor-OTR, prostaglandin (PGF)2α receptor-PGFR, muscarinic receptor-MAChR and estrogen receptor (ER) isoforms α and β. Levels of estradiol and progesterone in serum were determined by enzyme-linked immunoassay (ELISA). Ex-vivo contraction study revealed the force of uterine contraction increased with increasing doses of MPE. In addition, expression of CaM, MLCK, SERCA, Gα, Gβ, IP3K, OTR, PGF2α, MAChR, Erα and ERβ in the uterus increased with increasing doses of MPE. Serum analysis indicate that estradiol levels decreased while progesterone levels remained low at day-8 post-partum in rats receiving 250 and 500 mg/kg/day MPE. These findings support the claims that MPE help to firm the uterus and pave the way for its use as a uterotonic agent after delivery.

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006663   ZIN S R M, KASSIM N M, MOHAMED Z, FATEH A H, ALSHAWSH M A (Anatomy Dep, Malaya Univ, Kuala Lumpur- 50603, Email: normadiah_mk@um.edu.my) : Potential toxicity effects of Anastatica hierochuntica aqueous extract on prenatal development of Sprague-Dawley rats. J Ethnopharmacol 2019, 245, 112180.

Anastatica hierochuntica (A. hierochuntica) is a plant consumed in folk medicine for the treatment of reproductive system related problems and metabolic disorders. It is of concern that the herb is commonly consumed by pregnant women towards the end of pregnancy to ease the process of labour, despite the lack of studies evaluating its safety. This study aimed to investigate the potential toxicity effects of A. hierochuntica in pregnant Sprague-Dawley rats and their developing foetuses. Experiments were conducted in accordance to the Organisation for Economic Co-operation and Development guideline 414. Animals were randomly divided into four groups (n = 10 females per group): negative control (received the vehicle only), experimental animals received 250, 500, and 1000 mg/kg A. hierochuntica aqueous extracts (AHAE), respectively. Treatment was administered daily by oral gavage from gestational day (GD) 6–20, and caesarian section performed on GD21. There were significant reduction in the corrected maternal weight gain of dams and body weight of foetuses in the lowest and highest dose of AHAE-treated animals compared to the control. These findings were associated with the increase in anogenital distance index and multiple congenital anomalies observed in some of the offspring. On the other hand, rats treated with 500 mg/kg showed higher embryonic survival rate with absence of significant treatment-related effect. Findings showed that highest and lowest doses of AHAE have prenatal toxicity effects in SD rats. Therefore, AHAE is potentially harmful to the developing foetuses especially when consumed during the period of implantation and organogenesis. As for the rats treated with 500 mg/kg AHAE, there was no significant treatment-related effect. Hence, we postulate that this finding suggests that the disruption on the hormonal regulation could have been compensated by negative feedback response. The compensated effects of AHAE at 500 mg/kg and the presence of lowest observed adverse effect level (LOAEL) at 250 mg/kg has resulted in a non-monotonous dose response curve (NMDRC), which complicates the determination of the value of no-observed-adverse effect level (NOAEL). 

66 ref

006597   HWANG S-H, LORZ L R, YI D-K, NOH J K, YI Y-S, CHO J-Y (Integrative Biotechnology Dep, Sungkyunkwan Univ, Suwon- 16419, Email: sohyun031195@naver.com) : Viburnum pichinchense methanol extract exerts anti-inflammatory effects via targeting the NF-κB and caspase-11 non-canonical inflammasome pathways in macrophages. J Ethnopharmacol 2019, 245, 112161.

Viburnum pichinchense Benth. Mainly found in Ecuador and Colombia has been ethnopharmacologically utilized as a remedy for various female disorders with kidney inflammation and uterine relaxant. The pharmacological activity of Viburnum pichinchense has never been studied, therefore, this study explored anti-inflammatory activity of Viburnum pichinchense methanol extract (Vp-ME). Anti-inflammatory activities of Vp-ME were evaluated in lipopolysaccharide (LPS)-stimulated RAW264.7 cells and HCl/EtOH-induced gastritis mice by MTT assay, nitric oxide (NO) production assay, semi-quantitative reverse-transcriptase-polymerase chain reaction (RT-PCR), luciferase reporter assay, Western blotting, and enzyme-linked immunosorbent assays (ELISA). Anti-inflammatory compounds in Vp-ME were identified by high performance liquid chromatography (HPLC). Vp-ME inhibited NO production in RAW264.7 cells stimulated with pam3CSK4, poly I:C or LPS and in LPS-stimulated peritoneal macrophages without cytotoxicity and downregulated mRNA expression of inflammatory enzymes, inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) and pro-inflammatory cytokines, tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and IL-6. The anti-inflammatory activity was accomplished by inhibiting nuclear factor-kappa B (NF-κB) transcriptional activation, upstream signaling molecules in the NF-κB pathway, and caspase-11 non-canonical inflammasome in RAW264.7 cells. Moreover, Vp-ME exhibited in vivo anti-inflammatory activity by ameliorating gastritis symptoms, inhibiting iNOS and IL-6 mRNA expression and IκBα activation in mice. HPLC analysis identified resveratrol, quercetin, luteolin, and kaempferol as the anti-inflammatory components in Vp-ME. This study demonstrated Vp-ME has the anti-inflammatory activity via targeting NF-κB and caspase-11 non-canonical inflammasome pathways in macrophage-mediated inflammatory responses, suggesting Vp-ME could be developed as anti-inflammatory ethnopharmacological remedies to prevent and treat inflammatory diseases.

42 ref

006623   OLORUKOOBA A B, ODOMA S (Pharmacology and Therapeutics Dep, Ahmadu Bello Univ, Zaria, Nigeria, Email: aolorukooba@gmail.com) : Elucidation of the possible mechanism of analgesic action of methanol stem bark extract of Uapaca togoensis pax in mice. J Ethnopharmacol 2019, 245, 112156.

Uapaca togoensis is a medicinal plant used traditionally in Africa for the treatment of rheumatism, epilepsy, cough, pneumonia, vomitting and fever. Previously, the analgesic activity of its methanol stem bark extract has been scientifically demonstrated. However, the mechanism responsible for this activity remains to be investigated. To elucidate the possible mechanism(s) through which the methanol stem bark extract of Uapaca togoensis (MEUT) exhibits analgesic activity in mice. Analgesic activity of MEUT was evaluated using acetic acid-induced abdominal writhing test in mice at doses of 250, 500 and 1000 mg/kg orally. For the mechanistic studies, mice were pre-treated with Naloxone (2 mg/kg), Atropine (1 mg/kg), Yohimbine (1 mg/kg), Glibenclamide (10 mg/kg), Prazosin (1 mg/kg) and Yohimbine (1 mg/kg) 15 min prior to MEUT (1000 mg/kg) administration, then assessed using AAWT 1 h later. Data was analysed using One way Anova followed by Bonferroni post hoc test. The extract (at the doses of 250, 500 and 1000 mg/kg) and morphine (10 mg/kg) significantly (p < 0.05) decreased the number of abdominal writhes. Naloxone (opioid receptor antagonist), Atropine (muscarinic receptor antagonist) and Glibenclamide (ATP-sensitive K+ channel blocker) significantly (p < 0.05) reversed the analgesic effect of MEUT. On the other hand, Prazosin and Yohimbine (α₁ and α₂ receptor antagonists respectively) had no effect on the analgesic action of MEUT. The results obtained from this study suggests the possible involvement of opioidergic, cholinergic and sensitive potassium ATP channel pathways in the analgesic activity of the methanol stem bark extract of Uapaca togoensis. 

40 ref

006584   FAN S, LIU C, JIANG Y, GAO Y, CHEN Y, FU K, YAO X, HUANG M, BI H (Sun Yat-sen Univ, Guangzhou- 510 006, Email: bihchang@mail.sysu.edu.cn) : Lignans from Schisandra sphenanthera protect against lithocholic acid-induced cholestasis by pregnane X receptor activation in mice. J Ethnopharmacol 2019, 245, 112103.

Cholestasis is a clinical syndrome caused by toxic bile acid retention that will lead to serious liver diseases. Ursodeoxycholic acid (UDCA) and obeticholic acid (OCA) are the only two FDA-approved drugs for its treatment. Thus, there is a clear need to develop new therapeutic approaches for cholestasis. Here, anti-cholestasis effects of the lignans from a traditional Chinese herbal medicine, Schisandra sphenanthera, were investigated as well as the involved mechanisms. Adult male C57BL/6J mice were randomly divided into 9 groups including the control group, LCA group, LCA with specific lignan treatment of Schisandrin A (SinA), Schisandrin B (SinB), Schisandrin C (SinC), Schisandrol A (SolA), Schisandrol B (SolB), Schisantherin A (StnA) and Schisantherin B (StnB), respectively. Mice were treated with each drug (qd) for 7 days, while the administration of lithocholic acid (LCA) (bid) was launched from the 4th day. Twelve hours after the last LCA injection, mice were sacrificed and samples were collected. Serum biochemical measurement and histological analysis were conducted. Metabolomics analysis of serum, liver, intestine and feces were performed to study the metabolic profile of bile acids. RT-qPCR and Western blot analysis were conducted to determine the hepatic expression of genes and proteins related to bile acid homeostasis. Dual-luciferase reporter gene assay was performed to investigate the transactivation effect of lignans on human pregnane X receptor (hPXR). RT-qPCR analysis was used to detect induction effects of lignans on hPXR-targeted genes in HepG2 cells. Lignans including SinA, SinB, SinC, SolA, SolB, StnA, StnB were found to significantly protect against LCA-induced intrahepatic cholestasis, as evidenced by significant decrease in liver necrosis, serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP) activity. More importantly, serum total bile acids (TBA) and total bilirubin (Tbili) were also significantly reduced. Metabolomics analysis revealed these lignans accelerated the metabolism of bile acids and increased the bile acid efflux from liver into the intestine or feces. Gene analysis revealed these lignans induced the hepatic expressions of PXR-target genes such as Cyp3a11 and Ugt1a1. Luciferase reporter gene assays illustrated that these bioactive lignans can activate hPXR. Additionally, they can all upregulate hPXR-regulate genes such as CYP3A4, UGT1A1 and OATP2.

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006579   DUBOST J-M, LAMXAY V, KRIEF S, FALSHAW M, MANITHIP C, DEHARO E (Federal Univ in Toulouse, UPS, France, Email: eric.deharo@ird.fr) : From plant selection by elephants to human and veterinary pharmacopeia of mahouts in Laos. J Ethnopharmacol 2019, 244, 112157.

To what extent has animal observation contributed to the development of human pharmacopeias? We approach this question here through the study of mahouts’ knowledge regarding the responses by elephants to their health problems, and the human medicinal uses of plants and the care of domestic animals that result from their observations. 32 mahouts were interviewed in Thongmyxay district and 28 at the Elephant Conservation Center in the province of Xayabury. Interviews focused on the elephants’ diet, health problems, plant items they consume in particular physiological or pathological contexts and the treatments that mahouts provide them. For each plant mentioned, the part of the plant consumed and mode of preparation and administration if used by mahouts were recorded. Species samples were then collected and later identified by specialists. 114 species were recorded as being consumed by elephants during interviews with mahouts and forest outings with them to collect samples. Twenty species were identified as used by elephants in particular pathological conditions or physiological states. According to interviewed mahouts, the consumption of certain plants improves the health of the elephant. We observed clear convergences between the observations interpreted by the mahouts as self-medication behaviour from elephants and their own medicinal practices (for human and veterinary purposes). Beyond a mere reproduction of elephant self-medication behaviours observed, the human or veterinary medicinal cares derived from these observations are the result of complex arrangements integrating all available medicinal and conceptual resources into elaborate preparations.

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006587   GAWRON G, KRZYCZKOWSKI W, LEMKE K, OIDAK A, KADZINSKI L, BANECKI B (Molecular and Cellular Biology Dep, Medical Univ of Gdansk, Gdansk, Poland, Email: leszek.kadzinski@biotech.ug.edu.pl) : Nigella sativa seed extract applicability in preparations against methicillin-resistant Staphylococcus aureus and effects on human dermal fibroblasts viability. J Ethnopharmacol 2019, 244, 112135.

Nigella sativa L. seed extracts and oils have been embraced by traditional medicine of cultures inhabiting Middle East and North Africa for centuries. Among other uses, it has been applied against dermatitis and eczema often worsened by staphylococcal colonization of the skin. The study was conducted to evaluate applicability of N. sativa seed extract in antibacterial skin formulations by examination of its activity against methicillin-resistant Staphylococcus aureus as well as cytotoxicity against human dermal fibroblasts. Two variants of N. sativa seed extract containing 9.91 and 2.10 % of thymoquinone were prepared by supercritical carbon dioxide extraction. The extracts and standards of their major volatile ingredients; thymoquinone, thymol, p-cymene alongside with the reference antiseptics; chlorquinaldol and a combination of amylmetacresol with 2,4-dichlorobenzyl alcohol were subjected to evaluation of antibacterial efficacy against a collection of Staphylococcus aureus strains. The preparation based on Vaseline containing 1% of N. sativa extract was applied on Mueller-Hinton agar plates and its ability to inhibit S. aureus growth was examined. The MTT assay was employed to study cytotoxic effects of the thymoquinone-rich N. sativa seed extract against HDFa fibroblasts. N. sativa seed extract and thymoquinone have shown potent bacteriostatic and bactericidal effect against Staphylococcus aureus, including methicillin-resistant strains (MRSA) isolated in Poland. Results suggest that N. sativa seed extract activity against S. aureus should mainly be attributed to thymoquinone, which was effective in concentrations of 4–16 μg/ml. Regarding the activity against S. aureus, thymoquinone was more efficient than a combination of amylmetacresol with 2,4-dichlorobenzyl alcohol and comparable to chlorquinaldol. The Vaseline-based preparation containing N. sativa extract caused growth inhibition comparable to an equally concentrated DMSO solution of the extract. The IC50 of N. sativa extract against HDFa fibroblast was determined at 0.2 mg/ml, which was 2-fold higher than the average MIC and MBC of the extract against S. aureus. The observed effectiveness of N. sativa seed extracts against bacteria was found to be dominantly dependent on concentration of thymoquinone. Its efficiency against Staphylococcus aureus isolates as well as results of cytotoxicity examination against human dermal fibroblasts indicate on its applicability as an antibacterial agent for topical use and motivates further research in this area.

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006574   CHEN R, WANG J, ZHAN R, ZHANG L, WANG X (Guangdong Pharmaceutical Univ, Guangzhou- 510 006, Email: 975201721@qq.com) : Fecal metabonomics combined with 16S rRNA gene sequencing to analyze the changes of gut microbiota in rats with kidney-yang deficiency syndrome and the intervention effect of You-gui pill. J Ethnopharmacol 2019, 244, 112139.

A myriad of evidence have shown that kidney-yang deficiency syndrome (KYDS) is associated with metabolic disorders of the intestinal microbiota, while TCMs can treat KYDS by regulating gut microbiota metabolism. However, the specific interplay between KYDS and intestinal microbiota, and the intrinsic regulation mechanism of You-gui pill (YGP) on KYDS’ gut microbiota remains largely unknown so far. In the present study, fecal metabonomics combined with 16S rRNA gene sequencing analysis were used to explore the mutual effect between KYDS and intestinal flora, and the intrinsic regulation mechanism of YGP on KYDS's gut microbiota. Rats' feces from control (CON) group, KYDS group and YGP group were collected, and metabolomic analysis was performed using ¹H NMR technique combined with multivariate statistical analysis to obtain differential metabolites. Simultaneously, 16S rRNA gene sequencing analysis based on the Illumina HiSeq sequencing platform and ANOVA analysis were used to analyze the composition of the intestinal microbiota in the stool samples and to screen for the significant altered microbiota at the genus level. After that, MetaboAnalyst database and PICRUSt software were apply to conduct metabolic pathway analysis and functional prediction analysis of the screened differential metabolites and intestinal microbiota, respectively. What's more, Pearson correlation analysis was performed on these differential metabolites and gut microbiota. Using fecal metabonomics, KYDS was found to be associated with 21 differential metabolites and seven potential metabolic pathways. These metabolites and metabolic pathways were mainly involved in amino acid metabolism, energy metabolism, methylamine metabolism, bile acid metabolism and urea cycle, and short-chain fatty acid metabolism. Through 16S rRNA gene sequencing analysis, we found that KYDS was related to eleven different intestinal microbiotas. These gut microbiota were mostly involved in amino acid metabolism, energy metabolism, nervous, endocrine, immune and digestive system, lipid metabolism, and carbohydrate metabolism. Combined fecal metabonomics and 16S rRNA gene sequencing analysis, we further discovered that KYDS was primarily linked to three gut microbiotas (i.e. Bacteroides, Desulfovibrio and [Eubacterium]_coprostanoligenes_group) and eleven related metabolites (i.e. deoxycholate, n-butyrate, valine, isoleucine, acetate, taurine, glycine, α-gluconse, β-glucose, glycerol and tryptophan) mediated various metabolic disorders (amino acid metabolism, energy metabolism, especially methylamine metabolism, bile acid metabolism and urea cycle, short-chain fatty acid metabolism. nervous, endocrine, immune and digestive system, lipid metabolism, and carbohydrate metabolism). YGP, however, had the ability to mediate four kinds of microbes (i.e. Ruminiclostridium_9, Ruminococcaceae_UCG-007, Ruminococcaceae_UCG-010, and uncultured_bacterium_f_Bacteroidales_S24-7_group) and ten related metabolites (i.e. deoxycholate, valine, isoleucine, alanine, citrulline, acetate, DMA, TMA, phenylalanine and tryptophan) mediated amino acid metabolism, especially methylamine metabolism, bile acid metabolism and urea cycle, short-chain fatty acid metabolism, endocrine, immune and digestive system, and lipid metabolism, thereby exerting a therapeutic effect on KYDS rats. Overall, our findings have preliminary confirmed that KYDS is closely related to metabolic and microbial dysbiosis, whereas YGP can improve the metabolic disorder of KYDS by acting on intestinal microbiota. Meanwhile, this will lay the foundation for the further KYDS's metagenomic research and the use of intestinal microbiotas as drug targets to treat KYDS.

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006658   ZHAO L, LIU S, WANG M, ZHI M, GENG S, HOU C, WANG W, ZHAO D (Hebei Medical Univ, Shijiazhuang- 050 017, Email: zhaodingd@163.com) : Berberine restored nitrergic and adrenergic function in mesenteric and iliac arteries from streptozotocin-induced diabetic rats. J Ethnopharmacol 2019, 244, 112140.

Perivascular neuropathy was reported to involve in the vascular disorders associated with diabetes. The dried rhizomes of Coptis chinensis Franch. (Latin name: Coptidis Rhizoma; common name: Huang Lian in China), used frequently in Traditional Chinese medicine to treat diabetes (Xiaoke), have been confirmed to possess beneficial effects on diabetic peripheral neuropathy by modern clinical and pharmacological studies. Berberine (BBR), the main effective component of Huang Lian in the treatment of diabetes, is reported to ameliorate diabetic central and peripheral neuropathy. However, the effects of BBR on nerve function of mesenteric and iliac arteries are unclear. To investigate the effects of BBR on the diabetes-induced changes in nitrergic and adrenergic function in mesenteric and iliac arteries. In this study, the animals were randomized into three groups: control rats, diabetic rats, and diabetic rats gavaged with BBR. We established diabetic rat model using intraperitoneal injection of streptozotocin (STZ, 55 mg kg⁻¹). Two weeks after model establishment, those in the BBR-treated groups were gavaged with berberine chloride (Sichuan Xieli Fharmaceutical. Co., Ltd; 200 mg·kg⁻¹·day⁻¹) diluted in distilled water for another 2 weeks. The superior mesenteric artery and iliac artery were excised. Electric field stimulation (EFS) was used to induce arterial vasoconstriction and explore (1) the diabetes-induced changes in neurogenic function of the superior mesenteric artery and iliac artery; (2) the effects of BBR on neurovascular dysfunction in the early stage of STZ-induced diabetic rats. Nitric oxide (NO) and noradrenaline (NA) released from the nitrergic and adrenergic nerves were quantified using fluorescence assays and ELISA, respectively. EFS induced frequency-dependent vasoconstrictions in both superior mesenteric and iliac artery, and the contractile responses of arteries were abolished by 0.1 μmol·L⁻¹ tetrodotoxin (TTX), or inhibited by 1 μmol·L⁻¹ phentolamine or increased by 0.1 mmol·L⁻¹ N^ω-Nitro-L-arginine methyl ester hydrochloride (L-NAME). In superior mesenteric artery, but not in iliac artery, the changes of contractile responses with L-NAME were significantly decreased in diabetic rats, and NO release was less also. In contrast, in iliac artery of diabetic rats, but not in superior mesenteric artery, the changes of contractile responses with phentolamine were increased, and NA release was increased significantly. All these changes in diabetic rats on both superior mesenteric artery and iliac artery were reversed by treated with BBR. In the STZ-induced early diabetic rats, neural control of mesenteric and iliac vasomotor tone are altered differently. The diminished nitrergic nerve in superior mesenteric artery and enhanced adrenergic nerve in iliac artery both contributed to increased vasocontrictor responses. All these changes in diabetic rats were reversed by BBR, suggesting a novel mechanism of BBR in balance of neural regulation of vascular tone.

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006659   ZHAO Q, REN X, CHEN M, YUE S-J, ZHANG M-Q, CHEN K-X, GUO Y-W, SHAO C L, WANG C-Y (Ocean Univ of China, Qingdao- 266 003, Email: shaochanglun@ouc.edu.cn) : Effects of traditional Chinese medicine formula Le-Cao-Shi on hepatitis B: In vivo and in vitro studies. J Ethnopharmacol 2019, 244, 112132.

Formula Le-Cao-Shi (LCS) is a traditional Chinese medicine (TCM), which has long been used as a folk remedy against hepatitis B in China. The present study was conducted to evaluate the anti-hepatitis B effects of aqueous extract of LCS in vivo and in vitro. we investigated the anti-HBV effects of LCS in vivo and in vitro with duck hepatitis B model and HepG2.2.15 cell line model, respectively. The serologic and cellular biomarkers and the histopathological changes were examined. By a duck hepatitis B model, the extract of LCS was found to restrain the expressions of duck hepatitis B surface antigen (DHBsAg), hepatitis B e antigen (DHBeAg), and HBV-DNA (DHBV-DNA). Moreover, LCS could decrease the levels of aspartate and alanine aminotransferases (AST and ALT) and ameliorate duck liver histological lesions. Correspondingly, in a HepG2.2.15 cellular model, LCS could also significantly inhibit the secretions of HBsAg and HBeAg. LCS exerted potent anti-hepatitis effects against the infection of HBV. The above results demonstrated the first-hand experimental evidences for the anti-hepatitis B efficiency of LCS. Our study provides a basis for further exploration and development of this promising compound prescription to treat hepatitis B disease.

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006661   ZHENG H, WANG X, ZHANG Y, CHEN L, HUA L, XU W (Fujian Univ of Traditional Chinese Medicine, Fujian- 350 122, Email: zhyfjtcm@163.com) : Pien-Tze-Huang ameliorates hepatic fibrosis via suppressing NF-κB pathway and promoting HSC apoptosis. J Ethnopharmacol 2019, 244, 111856.

Pien Tze Huang (PZH), a Chinese herbal formula, has various forms of pharmacological activity including anti-inflammation, liver protection and anti-tumor. To investigate the anti-hepatic fibrosis effect of PZH and its potential mechanisms on experimental animal model of hepatic fibrosis and hepatic stellate cell (HSC). Rats were intraperitoneally administered with CCl₄ to induce hepatic fibrosis and were simultaneously treated with PZH. Liver pathology were observed by hematoxylin and eosin (H&E) staining and Masson staining. Serum pro-inflammatory cytokines were determined by enzyme-linked immunosorbent assay (ELISA) kits. Moreover, the effects of PZH on HSC proliferation and apoptosis were assessed via MTT, flow cytometry and immunofluorescence analysis. Nuclear factor-κB (NF-κB) pathway activation and Bcl-2 family members were evaluated by reverse transcription quantitative polymerase chain reaction (real-time PCR) and western blotting. PZH significantly alleviated CCl₄-induced liver injury, inflammation and fibrogenesis in rats. PZH also markedly decreased the production of hepatic-fibrosis biomarker, including ALT, AST, IV collagen and PCIII (Procollagen III), as well as inflammatory cytokines such as IL-1β, IL-6 and TNF-α. Importantly, PZH strongly inhibited HSC proliferation correlated with cell apoptosis induction as evidenced by modulating Bcl-2 family members and caspase activity. Moreover, PZH administration significantly increased the expression IκB-α, an inhibitor of NF-κB and suppressed expression of anti-apoptotic genes (Bcl-2, etc.). Collectively, these results suggested that PZH could promote HSC apoptosis through inhibiting NF-κB signaling pathway. Our study demonstrates that PZH ameliorates hepatic fibrosis and inflammation, chiefly through suppressing the NF-κB pathway and promoting HSC apoptosis. PZH is more likely to be a promising antifibrotic agent in chronic disease.

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006572   CHEN B, LI J, XIE Y, MING X, LI G, WANG J, LI M, LI X, XIONG L (Integrative Medicine on Pediatrics Dep, Shanghai Jiao Tong Univ, Shanghai- 200 127, Email: lijijun523@163.com) : Cang-ai volatile oil improves depressive-like behaviors and regulates DA and 5-HT metabolism in the brains of CUMS-induced rats. J Ethnopharmacol 2019, 244, 112088.

Cang-ai volatile oil (CAVO) is a traditional Chinese medicine (TCM) inhalational preparation for the treatment of some depressive and emotive disorders. This research aimed to evaluate the efficiency and possible mechanism of intranasal CAVO administration on depression in chronic unpredictable mild stress (CUMS)-induced rats compared to lavender volatile oil (LVO) treatment after CUMS exposure and bilateral olfactory bulb impairment (OBI) in rats. Forty depressive-like model rats induced by CUMS were evaluated by the forced swim test (FST), open field test (OFT), and sucrose preference test (SPT). The model rats were divided into five groups: CUMS (n = 8), CAVOh + CUMS (n = 8), CAVOl + CUMS (n = 8), LVO + CUMS (n = 8), and OBI + CAVO + CUMS (n = 8). The CUMS-induced rats were treated for a period of 4 weeks. The other healthy rats were regarded as the control (CTR, n = 8) subjects. The levels of serotonin (5-HT) and dopamine (DA) and their respective metabolites homovanillic acid (HVA) and 5-hydroxyindol acetic acid (5-HIAA) were measured in brain tissue homogenates of CUMS-induced rats using enzyme-linked immunosorbent assay (ELISA). CAVO ameliorated depressive-like behaviors (p < 0.05). The levels of DA in the CUMS group were lower than those in the CTR and CAVOh groups (**p < 0.01 and *p < 0.05). The levels of HVA were lower in the CUMS group than in the CTR, LVO, OBI + CAVOh and CAVOh groups (**p < 0.01 and *p < 0.05) and lower in the OBI + CAVOh group than in the CAVOh group (**p < 0.01). The levels of 5-HT in the CUMS group were lower than those in the CTR and CAVOh groups (**p < 0.01). The levels of 5-HIAA were lower in the CUMS and OBI + CAVOh groups than in the CTR, LVO and CAVOh groups (**p < 0.01). CAVO can improve depressive-like behaviors concomitant with the regulation of DA and 5-HT metabolism in the brains of CUMS-induced rats.

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006575   CHENG C-F, LIN J C-F, TSAI F-J, CHEN C-J, CHIOU J-S, CHOU C-H, LI T-M, LIN T-H, LIAO C-C, HUANG S-M (China Medical Univ, Taichung, Taiwan, Email: jimmy.bbww@hotmail.com) : Protective effects and network analysis of natural compounds obtained from Radix dipsaci, Eucommiae cortex, and Rhizoma drynariae against RANKL-induced osteoclastogenesis in vitro. J Ethnopharmacol 2019, 244, 112074.

Osteoporosis is one of the most common bone diseases; it is characterized by bone loss and is a risk factor for hip fracture. Chinese herbal medicines (CHMs) and their related natural compounds have been used for treating many diseases, including bone diseases, since ancient times in China and are regarded as a cost-effective complementary therapy. The goal of this study was to investigate the osteoprotective mechanisms of these three Chinese herbs and their related natural compounds. The effects of CHMs and related natural compounds on RANKL-induced osteoclastogenesis in vitro were investigated. A network pharmacology method was applied to study CHM-related natural compounds and their osteoporosis targets. In addition, their effect on RANKL-induced osteoclastogenesis in RAW264.7 cells was also investigated in vitro. Radix dipsaci, Eucommiae cortex, and Rhizoma drynariae exhibited protective effects against mortality in hip fracture patients. Furthermore, these three herbs inhibited RANKL-induced TRAP activities and reduced the expression of bone resorption-related genes in RAW264.7 cells. Network analysis of natural compound (ingredient)-target interactions identified 11 natural compounds. Signal pathway analyses suggested that these compounds may target cytokine-cytokine receptor interactions, including RANKL-induced osteoclastogenesis. Five novel natural compounds exhibited reduced RANKL-induced TRAP activities and bone resorption-related gene expression. The clinically used CHMs, Radix dipsaci, Eucommiae cortex, and Rhizoma drynariae, and natural compounds obtained from them may suppress RANKL-induced osteoclastogenesis in vitro. 

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006566   BRITO V G B, CHAVES-NETO A H, BARROS T L D, OLIVEIRA S H P (Basic Sciences Dep, São Paulo State Univ, SP, Brazil, Email: victorbalera@gmail.com) : Soluble yerba mate (Ilex Paraguariensis) extract enhances in vitro osteoblastic differentiation of bone marrow-derived mesenchymal stromal cells. J Ethnopharmacol 2019, 244, 112131.

Yerba mate (Ilex paraguariensis) consumption has been associated with beneficial effects on bone health. The purpose of this study was to evaluate the mechanism by which soluble yerba mate (SYM) stimulates osteoblast differentiation of bone marrow-derived mesenchymal stromal cells (BM-MSCs). BM-MSCs from male Wistar rats were induced towards osteoblastic differentiation with different concentrations of SYM (10, 20, and 50 μg/mL). Osteoblastic differentiation was evaluated by measuring proliferation rates, alkaline phosphatase activity, MMP-2 activity, mineralization, and gene expression of Runx2, Osterix, β-catenin (Catnb), collagen type I (Col1a1), osteopontin (Opn), osteocalcin (Ocn), bone sialoprotein (Bsp), bone morphogenetic protein-2 (Bmp2), osteoprotegerin (Opg), and Rankl. We also analyzed cytokine production and MAP kinase pathways. SYM (10 μg/mL) did not show a cytotoxic effect and induced a slight increase in ALP activity; however, a great increase in mineralization was observed. SYM was also able to reduce TNF-α and IL-10 production; increase the expression of transcription factors Runx2, Osterix, and Catnb; and increase matrix proteins Opn, Bsp, Ocn, and Bmp2. We also observed a decrease in intracellular signaling of ERK, JNK, and p38 MAPK, which seemed to be related to the SYM response. Together, these results help to explain the promoting effect on osteoblast differentiation produced by a low SYM concentration. However, a higher SYM concentration presented deleterious effects, including cytotoxicity, decreased ALP activity, increased cytokine production, decreased bone marker gene expression, increased MAPK signaling, and significant mineralization reduction. In conclusion, our results suggest a concentration-specific direct stimulatory effect of SYM on osteoblastic differentiation in vitro. 

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006562   BALDIN V P, SCODRO R B D L, FERNANDEZ C M M, LEQUE A L, CALEFFI-FERRACIOLI K R, SIQUEIRA V L D, ALMEIDA A L D, GONCLAVES J E, CORTEZ D A G, CARDOSO R F (Estadual de Maringa Univ, Maringa- 87020-900, Email: vanessapbaldin@gmail.com) : Ginger essential oil and fractions against Mycobacterium spp. J Ethnopharmacol 2019, 244, 112095.

Zingiber officinale (ginger) is a perennial herbaceous plant native in tropical Asia and generally cultivated in most American tropical countries with widespread use in popular medicine. Ginger essential oil (GEO) has been reported to exhibit several biological activities, such as antimicrobial. The aim of this study was to determine the composition and the property of GEO and related fractions against Mtb and NTM, as well as their cytotoxicity. GEO was obtained by hydrodistillation and fractionation was performed. Chemical characterization of GEO and fractions were carried out by gas chromatography/mass spectrometry. The antimycobacterial activity was evaluated by resazurin microtiter assay plate and broth microdilution method for Mtb and NTM, respectively. The cytotoxicity in Vero cells was assessed by MTT colorimetric assay. The analyses showed 63 compounds in the GEO sample, characterized by a high number of monoterpenes and sesquiterpenes. GEO fractionation rendered 11 fractions (FR1 to FR11). GEO and fractions minimum inhibitory concentration ranged from 31.25 to >250 μg/mL against Mtb and from 15.6 to >250 μg/mL against NTM. GEO showed better activity against NTM, M. chelonae, and M. abscessus sub. massiliense, than the semi-pure fractions. One fraction (FR5), containing γ-eudesmol as the main compound, was the most active against Mtb and NTM. The GEO and semi-pure fractions cytotoxicity assay showed CC₅₀ 63.3 μg/mL, and 36.3–312.5 μg/mL, respectively. In general, GEO showed a mix of monoterpenes and sesquiterpenes and a better antimycobacterial activity than the semi-pure fractions. Cytotoxic effects of GEO and its fractions should be better investigated.

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006571   CHEGE B M, WAWERU M P, FREDRICK B, NYAGA N M (Medical Physiology Dep, Nairobi Univ, GPO- 30197-00100, Email: bmchege87@gmail.com) : The freeze-dried extracts of Rotheca myricoides (Hochst.) steane & mabb possess hypoglycemic, hypolipidemic and hypoinsulinemic on type 2 diabetes rat model. J Ethnopharmacol 2019, 244, 112077.

Rotheca myricoides (Hochst.) Steane & Mabb is a plant species used in traditional medicine for the management of diabetes in the lower eastern part of Kenya (Kitui, Machakos and Makueni Counties, Kenya) that is mainly inhabited by the Kamba community. This study investigated the antihyperglycaemic, antidyslipidemic and antihyperinsulinemic activity of the freeze-dried extracts of Rotheca myricoides (Hochst.) Steane & Mabb (RME) in an animal model of type 2 diabetes mellitus.Type 2 diabetes was induced by dietary manipulation for 56 days via (high fat- high fructose diet) and intraperitoneal administration of streptozocin (30 mg/kg). Forty freshly-weaned Sprague Dawley rats were randomly assigned into the negative control (high fat/high fructose diet), low dose test (50mg/kg RME, high dose test (100mg/kg RME and positive control (Pioglitazone, 20mg/kg) groups. Fasting blood glucose and body weight were measured at weekly intervals. Oral glucose tolerance tests were performed on days 28 and 56. Lipid profile, hepatic triglycerides, fasting serum insulin levels and serum uric acid were determined on day 56. The RME possessed significant antihyperglycemic [FBG: 6.5 ± 0.11 mmol/l (negative control) vs. 4.62 ± 0.13 mmol/l (low dose test) vs. 5.25 ± 0.15 mmol/l in (high dose test) vs. 4.33 ± 0.09 mmol/l (positive control): p < 0.0001] and antihyperinsulinemic effects [1.84 ± 0.19 (negative control) vs. (0.69 ± 0.13 (low dose test) vs. (0.83 ± 0.17 (high dose test) vs. (0.69 ± 0.10 (positive control): F (3, 36) = 0.6421: p < 0.0001. The extracts also possessed significant antidyslipidemic effects [LDL levels: 3.52 ± 0.19 mmol/l (negative control) vs. 0.33 ± 0.14 mmol/l (low dose test) vs. 0.34 ± 0.20 mmol/l (high dose test) vs. 0.33 ± 0.01 mmol/l (positive control): p < 0.0001].RME significantly lowered plasma uric acid levels, as well as hepatic triglycerides and hepatic weights. Network pharmacology analysis indicated that the observed pharmacological effects are mediated via the modulation of Peroxisome proliferator-activated gamma receptor. The freeze dried extracts of Rotheca myricoides possessed significant antihyperglycemic and antidyslidemic effects. In addition it lowered serum uric levels, as well as hepatic triglycerides and hepatic weight. These results appear to validate the traditional use of this plant species in the management of diabetes mellitus.  

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006635   QIN D, ZHANG H, ZHANG H, SUN T, ZHAO H, LEE W-H, (Kunming Institute of Zoology, Kunming- 650 223, Email: leewh@mail.kiz.ac.cn) : Anti-osteoporosis effects of osteoking via reducing reactive oxygen species. J Ethnopharmacol 2019, 244, 112045.

 Osteoking is a Traditional Chinese Medicine consisting of seven types of medicinal herbs originated from Yi nationality and has been used in clinic to treat bone diseases for thousands of years in China. Osteoking shows excellent clinical therapeutic effects on osteoporosis, but it is not clear whether Osteoking could exhibit beneficial effects against osteoporosis via reducing reactive oxygen species (ROS). To explore whether the protective effects of Osteoking on osteoporosis related to ROS, we investigated the effects of Osteoking on osteogenesis differentiation under oxidative stress. The ovariectomized (OVX) osteoporosis model was established by ovarian surgery, and Osteoking was orally administrated for 84 days. Then the pathogenesis changes of femur were analyzed by Hematoxylin and eosin (H&E) and Masson's trichrome staining. The levels of ROS, malondialdehyde (MDA)and superoxide dismutase (SOD) from rats' serum were further measured. In vitro, mouse pre-osteoblastic MC3T3-E1 cells pre-treated with or without 0.25 mM tert-butyl hydroperoxide (t-BHP) for 2 h were cultured and treated with different dilutions of Osteoking or 20 μM N-Acetyl-L-cysteine for another 24 h, respectively. The intracellular ROS production and markers of oxidative damage of the MC3T3-E1 cells were determined using corresponding kits, respectively. The expressions of alkaline phosphatase (ALP), collagen type I, osteoprotegerin (OPG), TGF-β1, β-catenin, receptor activator of nuclear factor-κB ligand (RANKL) and interleukin-6 (IL-6) were further analyzed by qRT-PCR and western blotting upon treatment. Our results showed that Osteoking significantly improving trabecular microstructure by promoting collagen fiber repair and new bone or cartilage regeneration was demonstrated in OVX osteoporosis rat models by micro-CT analysis and histological staining results. Osteoking supplementation reduced the levels of ROS and MDA in OVX rat serum and increased SOD activities. In addition, Osteoking could also up-regulate the proteins expression levels of Runx2, osteocalcin (BGP) and osteoprotegerin (OPG) but reducing the expression of tartrate-resistant acid phosphatase (TRAP). In vitro, Osteoking could effectively inhibit the t-BHP-induced intracellular excessive ROS production and protect cells from oxidative stress in mouse pre-osteoblastic MC3T3-E1 cells. Meanwhile, the mRNA expressions of ALP, collagen type I, OPG, TGF-β1 and β-catenin were also up-regulated whereas the RANKL and IL-6 were down-regulated in Osteoking-treated MC3T3-E1 cells. A novel therapeutic mechanism of Osteoking on osteoporosis reveals by present investigation. Clinic effects of Osteoking to treat osteoporosis are closely related to its ability to reduce oxidative stress.

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006606   KIM S-B, SEO Y-S, KIM H S, LEE A Y, CHUN J M, MOON B C, KWON B-I (Herbal Medicine Research Div, Korea Institute of Oriental Medicine, Daejeon- 34054, Email: kbi34812@sangji.ac.kr) : Anti-asthmatic effects of lepidii seu Descurainiae semen plant species in ovalbumin-induced asthmatic mice. J Ethnopharmacol 2019, 244, 112083.

Lepidii seu Descurainiae Semen (LDS) is used as a traditional herbal medicine in northeast Asia, mainly in Korea, Japan, and China to treat lung disorders including coughs and phlegm caused by acute and chronic airway inflammation. Recently, interest regarding health problems incurred by air pollution has rapidly grown. Herbal medicines are being considered as alternative agents to treat various diseases. In the present study, we evaluated and compared the anti-inflammatory effects of LDS, which is derived from Lepidium apetalum Willd. extracts (LAE) and Descurainia sophia (L.) Webb ex Prantl extracts (DSE), on allergic airway inflammation. We established an ovalbumin-induced asthmatic mouse model to evaluate the efficacy of LDS extracts. We performed histological examination and measured relevant inflammatory mediators and cells in bronchoalveolar lavage fluid and lung. Furthermore, we conducted an in vitro T helper 2 (Th2) polarization assay, flow cytometry, and western blot analysis. Asthmatic phenotypes were attenuated by LDS extract treatments. LDS extract administration significantly reduced mucus production, inflammatory cell infiltration into airways, and eosinophil activation. Furthermore, LDS extracts reduced the expression of type 2 cytokines and inhibited differentiation and activation of Th2 cells. LDS alleviated eosinophilic inflammation by inhibiting Th2 cell differentiation, and DSE was more effective in attenuating allergic lung inflammation than LAE.

45 ref

006650   TOIT A D, KOOY F V D (North-West Univ, Potchefstroom- 2520, Email: annekedutoit00@gmail.com) : Artemisia afra, a controversial herbal remedy or a treasure trove of new drugs?. J Ethnopharmacol 2019, 244, 112127.

Artemisia afra is one of the most widely used herbal remedies in South Africa. This highly aromatic shrub is used to treat various disorders including coughs, colds, influenza, and malaria. Due to the long tradition of use and popularity of A. afra, it has been successfully commercialised and can currently be bought from various internet stores and pharmacies. The most notable indication is for the prophylaxis and treatment of Plasmodium falciparum infections. In 2013, the Medicine Control Council (MCC) of South Africa banned the sale of A. afra for the treatment of malaria because it lacks scientific evidence of efficacy. This resulted in a lawsuit being filed in 2017 against the MCC by an herbal company which claimed that artemisinin was responsible for A. afra's antiplasmodial activity. At the time, no scientific literature reported that A. afra contained artemisinin. This review aims to collate all available scientific literature regarding the phytochemistry and biological activity, focusing on antimalarial activity, of A. afra published from 2009 to 2019 and follows on our earlier review, which covered all literature until 2009. All scientific literature in English published between 2009 and June 2019 were retrieved from scientific databases (Scifinder scholar, Web of Science, Scopus, PubMed, Google scholar) and a number of books regarding medicinal plants in South Africa were also consulted. In the last decade very few compounds have been identified in A. afra, none of which were novel compounds. Based on all the tests that have been conducted using extracts and compounds of A. afra in a disparate variety of in vitro and in vivo bioassays, the results indicate only weak biological activity. The activity of extracts, and in some cases pure compounds, exhibited IC₅₀ or MIC values of 1000–10 000 fold less active than the positive controls. In contrast, and quite surprisingly, two randomised controlled trials were recently conducted (Schistosoma mansoni and Plasmodium falciparum infected patients) and although criticised based on design, execution, statistical analysis and ethical concerns, showed remarkably positive results. Pre-clinical in vitro and in vivo animal experiments failed to yield any promising drug leads. However, if the recent randomised controlled trials can be independently replicated in well-designed and executed clinical trials it might indicate that A. afra contain powerful ‘prodrugs’. Future research on A. afra should therefore focus on reproducing the randomised controlled trials and on artificially metabolising A. afra extracts/compounds in order to identify the presence of any ‘prodrugs’.

65 ref

006580   ELOUADDARI A, ELAMRANI A, MOUTIA M, OUBRIM N, HABTI N, EDDINE J J (Hassan Univ, Casablanca, Morocco) : Chemical composition and evaluation of antioxidant, antimicrobial and cytotoxic activities of Moroccan Cladanthus mixtus essential oil and extracts. J Essent Oil Bear Plants 2019, 22(6), 1450-66.

The methanolic and deodorized aqueous extracts of the Moroccan Cladanthus mixtus collected from Bouznika were studied for its antioxidant, antimicrobial and cytotoxic activities. The essential oil was isolated by hydrodistillation and analyzed using GC/FID and GC/MS. The major component found was 2- methyl-2, trans-butenyl methacrylate (34 %). The essential oil showed good antioxidant activity for DPPH (IC₅₀=0.342±0.002 mg/mL) and a strong antimicrobial activity against all the tested microorganisms. Although, the deodorized aqueous extract had a high amount of phenols (38.2±0.4 mg GAE/g), it has shown no inhibiting effect against the growth of tested bacterial strains. The methanolic extract of C. mixtus which was characterized with higher flavonoid content (3.2 ±0.01 mg QE/g) was analyzed by the HPLC and showed the presence of gallic, vanillic, caffeic and syringic acids and catechin. For the methanolic extract, higher antioxidant activity was recorded than the deodorized aqueous extract using DPPH test (IC50 = 1.933 ± 0.048 mg/mL) and ABTS test (IC₅₀ = 2.030 ± 0.006 mg/mL). The essential oil showed good cytotoxic activity with an IC₅₀ = 85.27 μg/mL greater than that of the methanol extract (IC₅₀ = 125.5 μg/mL). The apigenin (42 , 5, 7,-trihydroxyflavone), a natural compound, isolated for the first time from C. mixtus, showed a great cytotoxic activity (IC₅₀ = 17.9 μg/mL). The 7,4'- dipropargyl ether, a semi synthetic derivative of apigenin obtained by alkylation reaction of apigenin, showed a lower cytotoxic activity (IC₅₀ =88 μg/mL) than apigenin. This present study also elucidated the role of hydroxyl groups at positions C7 and C4’on the cytotoxic activity.

6 illus, 6 tables, 50 ref

006636   QU Z-Y, JIN Y-P, CUI L-I, LI Y-I, REN Y, WANG H-C, WANG Y-P (Chinese Academy of Agricultural Sciences, Jilin- 130 112) : Chemical composition and cytotoxic activity of the essential oils from Orobanche cernua loefling whole plant. J Essent Oil Bear Plants 2019, 22(6), 1427-34.

Orobanche cernua is one of the species of Orobanche in Orobanchaceae family, which has been used in folk medicines for boost immunity and relieve swelling. The present study aimed to analyse the chemical constituents of the essential oils (EO) from O. cernua by GC/MS and evaluate the cytotoxic activity in vitro. Twenty-eight compounds, representing 83.41 % of the total EO were identified and the major volatile components were diethylhexyl adipate (35.34 %), followed by 2-methylheptane (12.65 %), α-cadinol (7.25 %), diethylphthalate (5.17 %), dibutyl phthalate (4.07 %), 3-methylheptane (3.51 %) and 1-tetradecanol (3.39 %). The EO cytotoxicity on the tumor cell lines (SH-SY5Y and BV-2) was determined by MTT assay and cell imaging, the results indicated that the potent cytotoxic effects with the IC₅₀ values (55.89 μg/mL) in SH-SY5Y cells but weak in BV-2 cells with the IC₅₀ (482.1 μg/mL) for 24 h, respectively. This paper is the first report of the chemical composition and cytotoxic activity of EO from O. cernua.

2 illus, 1 table, 32 ref

006617   MONDAL G, DEVI S D, KHAN Z A, LABALA R K, CHATTORAJ A (Animal Science Dep, Kazi Nazrul Univ, Asansol- 713 340, Email: asamanja.chattoraj@gmail.com) : Light, feeding and melatonin: An interplay in the appetite regulation in the gut of zebrafish (Danio rerio). J Endocrinol Reprod 2019, 23(2), 81-97.

Every physiological function, including feeding and energy homeostasis is vital for animal sustainability. Melatonin is the neuroendocrine transducer of circadian photoperiod and is able to synchronize these physiological functions. The present study demonstrates the daily variation in gut melatonin and mRNA expression of appetite regulating hormone [leptin, nesfatin-1, orexin, ghrelin and ghrelin o-acetyltransferase (goat)] in relation to Gastrointestinal Somatic Index (Ga-SI) in the gut of zebrafish (Danio rerio), under various lighting schedule- LD (12L:12D), LL (continuous Light), and DD (continuous dark)- and scheduled feeding. The result exhibited a change in the peak level of the Ga-SI according to different photic conditions. But no change in Ga-SI was found after melatonin treatment under normal photoperiod (LD). Peak expression of anorexigenic peptide hormone genes (leptin and nesfatin-1 ) were found with the highest level of Ga-SI but the highest-level mRNA expression of orexigenic peptide gene hcrt was at the time of highest feeding and ghrelin with goat were after 6 hr of highest Ga-SI. These patterns changed in continuous photic conditions. This result indicates light as the critical dominant factor and, may be through melatonin, it can modulate the components of appetite regulation in this peripheral organ. This finding supports the hypothesis about the “light pollution” and its silent desynchronization of feeding behavior and related physiological functions, thereby the biodiversity and the sustainability of organisms.

8 illus, 2 tables, 79 ref

006593   GUPTA C, NAIK S, KURIAN P, KALTHUR S, PALIMAR V, D’SOUZA A (Anatomy Dep, Kasturba Medical Coll, Manipal- 576 104, Email: chandnipalimar@gmail.com) : A morphological and morphometric study of distal end of ulna with its clinical implications. J Datta Meghe Inst Med Sci 2019, 14(4), 360-4.

The distal end of the ulna is of utmost anatomical importance for normal hand functioning. It can get damaged in various injuries such as avulsion of the ulnar styloid process or ulnar styloid triquetral impaction syndrome. Hence, the aim of this study is to measure the various dimensions of distal end of ulna and to look for its various morphological features. The study was conducted on fifty dry ulnae of unknown sex. Length ofstyloid process, width of fovea, and pole and height ofseat were measured. Various morphological features of the distal end of ulna were also noted down.  The mean length of styloid process, width of fovea, and pole were 0.55, 0.47, and 0.56 cm, respectively. The mean height of seat was 0.72 cm. In 59.57% of cases, the styloid process was straight and in 78.7% of cases, the tip of the styloid process was blunt. The most common shape of pole was kidney shape in 48% of cases. The results of this study will be useful for hand surgeons in dealing with distal ulnar injuries and even for reconstruction of the distal radioulnar joint with prosthesis of distal end of ulna.

4 illus, 3 tables, 8 ref