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Comparison of slow freezing and vitrification methods for Venda cockerel’s spermatozoa

Open Journal of Animal Sciences 2012年第3期2卷 204-210页

作者： Masindi L. Mphaphathi dibungi luseba Ben Sutherland Tshimangadzo L. Nedambale Agricultural Research Council Animal Production Institute Germplasm Conservation & Reproductive Biotechnologies Irene RSA Tshwane University of Technology Faculty of Science Department of Animal Sciences Pretoria RSA University of the Free State Department of Animal Wildlife and Grassland Sciences Bloemfontein RSA

An improvement in avian semen cryopreservation is essential and has the potential to improve the cryo-gene banking efficiency. This study compared two cryopreservation methods (slow freezing and vitrification) and the effect of different thawing/warming temperatures (5℃, 25℃ and 41℃) on Venda cockerel’s spermatozoa. Semen samples from Venda cockerels were diluted with modified Kobidil+ extender supplemented with 8% dimethyl sulfoxide. Semen from each ejaculate was stained with nigrosin/eosin for viability examination. The cryopreserved samples were either slow cooled in 0.25 mL straw or vitrified in a solid surface vitrification (SSV) device. Semen straw or cryovial was stored in liquid nitrogen container. The straw or cryovial with sperm was thawed or warmed at 5?C, 25?C and 41℃ and analysed by a Computer-Aided Sperm Analysis (CASA). There was a significant difference in live/normal sperm between the semen donors. Cockerels spermatozoa cryopreserved by slow freezing (43%) and thawed at 5?C had a significantly higher survival and motility rate compared to vitrification (2.5%) method. In conclusion, there was higher rate of live/normal morphology sperm. Cryopreservation process reduces sperm motility and velocity rate regardless of cryoprevervation method and thawing or warming temperatures. However, slow freezing was a better method to maintain motility of spermatozoa following cryopreservation.

关键词： Venda Cockerels Dimethyl Sulfoxide Sperm Motility Cryopreservation

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Cellular Antioxidant Activity and Peroxidase Inhibition of Infusions from Different Aerial Parts of Cassia occidentalis

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Journal of Biosciences and Medicines 2019年第4期7卷 83-94页

作者： Nadege Kabamba Ngombe Claude Ngolo Ngolo Didi Mana Kialengila Arvy Luzayana Wamba Patricia Mbombo Mungisthi Pascal Tshisekedi Tshibangu Pascal Kalenda T. dibungi Paul Tshilumbu Kantola Paulin Mutwale Kapepula Centre d’Etudes des Substances Naturelles d’Origine Végétale (CESNOV) Faculty of Pharmaceutical Sciences University of Kinshasa Kinshasa Democratic Republic of Congo Laboratory of Drug Analysis Faculty of Pharmaceutical Sciences University of Kinshasa Kinshasa Democratic Republic of Congo Laboratory of Experimental and Pharmaceutical Microbiology Faculty of Pharmaceutical Sciences University of Kinshasa Kinshasa Democratic Republic of Congo

Cassia occidentalis L. is widely used in the world in traditional medicine and especially in some African countries for the treatment of various diseases. The aim of this study was to report the microscopic features, the chromatographic fingerprints and the cellular antioxidant activity and the peroxidase inhibition of infusions from different parts of this plant. Microscopically, leaf can be characterized by cells of the spongy mesophyll and parenchyma numerous cluster crystals of calcium oxalate, paracytic stomata, isolated calcium oxalate cluster crystals, covering and glandular trichomes, scalariform vessels, polyedric starch granules, lignified fibers;flowers by abundant covering and glandular trichomes, spirally thickened vessels and associated parenchyma, abundant pollen grains. Seeds were characterized by pluricellular non-glandular trichomes, epidermis of the testa with underlying oil cells, parenchymatous layers of the testa, thicker-walled cells of the endosperm, pollen grain. Phytochemical analysis revealed the presence of phenolic acids, flavonoids, iridoids, tannins and terpenes. TLC fingerprints of different parts were different and characteristic. They showed the presence of glycosylated flavonoids and phenolic acids as main phytochemicals for flowers, leaves and seedpods. ABTS and DPPH assays showed that infusion extracts have the ability to scavenge free radicals connected with their IC50 values ranging from 21.43 ± 1.25 to 566.24 ± 176.7 mg·mL-1. All extracts showed a weaker capacity to scavenge DPPH radical. Aqueous extracts displayed high cellular antioxidant activity at the concentrations range of 1 - 20 μg·mL-1 using LO-12 on monocytes HL 60. Flower and leave extracts showed more efficient effects on extracellular ROS production. Phenolic compounds could be major contributors to antioxidant activity of infusions of Cassia parts. In MPO (Myeloperoxidase) direct technique, all infusion extracts exhibited a dose-dependent inhibitory effect on MPO

关键词： Cassia occidentalis HRP Manalaria Monocytes HL 60 MPO Nutraceutical

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Microscopic Characteristics, Chromatographic Profiles and Inhibition of Peroxidase Activity of the Leaves of Manihot esculentaand Manihot glaziovii, Consumed as Traditional Vegetables

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Journal of Biosciences and Medicines 2021年第9期9卷 59-73页

作者： Paulin Mutwale Kapepula Patricia Mbombo Mungitshi Dieudonné Tshitenge Tshitenge Thierry Franck Dieudonné Mumba Ngoyi Pascal dibungi T. Kalenda Monique Tits Michel Frédérich Nadege Kabamba Ngombe Ange Mouithys-Mickalad Centre d’Etudes des Substances Naturelles d’Origine Végétale (CESNOV) Faculty of Pharmaceutical Sciences University of Kinshasa Kinshasa Democratic Republic of the Congo Centre for Oxygen Research and Development (CORD) University of Liège Liège Belgium Faculty of Medicine University of Kinshasa Kinshasa Democratic Republic of the Congo Laboratory of Pharmacognosy Center for Interdisciplinary Research on Medicines (CIRM) University of Liège Liège Belgium

Methanolic extracts from the leaves of Manihot esculenta (Two cultivars) and Manihot glaziovii, consumed as traditional vegetables in DR. Congo was chemically characterized by Thin layer Chromatography and High Performance Liquid Chromatography. In vitro biochemical activities of extracts against Radical Oxidative Species (ROS) production were assessed in cellular models, on enzymes, Myeloperoxidase (MPO) and Horseradish Peroxidase (HRP) involved in inflammation. The microscopic analysis of the powder of leaves showed that each species displays specific and discriminating botanical microscopic features. Varieties of M. esculenta had a chemical fingerprint different from M. glaziovii. The majority of compounds were polyphenols, represented mainly by rutin, kaempferol-3-O-rutinoside, amentoflavone, phenolic acids such as gallic acid. All extracts exhibited high cellular antioxidant activity in the range of 0.1 to 10 μg·mL-1 using lucigenin with neutrophils, but a moderate cellular antioxidant activity ranging between 10 and 100 μg·mL-1 with DCFDA on HL60 monocytes. Extracts from Manihot leaves showed a pronounced inhibitory effect on the production of extracellular ROS, on HRP and myeloperoxidase activity. Cellular antioxidant activities, the inhibitory effect on HRP of extracts from M. glaziovii, M. esculenta cultivar Mwambu were significantly higher, but their inhibitory effect on the activity of MPO was lower than those of M. esculenta cultivar TEM 419. The biological activities of Manihot esculenta and Manihot glaziovii were well correlated to their phytochemicals that could justify their traditional use as vegetables, potential functional foods or nutraceutical resources and medicines.

关键词： Horseradish Peroxidase Kahemba Konzo Manihot esculenta Manihot glaziovii Myeloperoxidase Traditional Vegetable

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Investigation of the Quality of Antibiotics-Based Amoxicillin for Monitoring of Some Different Medicine Markets of Democratic Republic of Congo

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American Journal of Analytical Chemistry 2018年第8期9卷 366-385页

作者： Nicodème Kalenda Tshilombo Patient Ciza Hamuli Jérémie K. Mbinze Védaste Habyalimana dibungi T. Kalenda Don Jethro Mavungu Pierrot Mwamba Philippe Hubert Roland Djang’eing’a Marini Laboratory of Analytical Chemistry CIRM Department of Pharmacy Liege University Liège Belgium Laboratory of Chromatography Faculty of Pharmaceutical Sciences University of Kinshasa Kinshasa Democratic Republic of Congo Laboratory of Drug Analysis Department of Galenic Pharmacy University of Kinshasa Kinshasa Democratic Republic of Congo Medical Procurement and Production Division Rwanda Biomedical Center Kigali Rwanda Laboratory of Pharmacognosy CIRM Department of Pharmacy Liege University Liège Belgium Ecole Régionale Postuniversitaire d’Aménagement et de Gestion Intégrée des Forêts et Territoires Tropicaux University of Kinshasa Kinshasa Democratic Republic of Congo Department of Pharmacy Galenic University of Lubumbashi Lubumbashi Democratic Republic of Congo

In order to combat the counterfeiting of drugs, adapted HPLC analytical USP methods were applied to evaluate the quality of the amoxicillin (with or without potassium clavulanate) powder for suspension sold in some Congolese markets. The adaptation has been done by modifying the column dimensions and adjusting the flow rate. According to the intended deployment of these methods in Democratic Republic of Congo (DRC), 3 factors (analyst, day and equipment) were involved in the validation step while applying the classic total error measurement approach with an accuracy profile as decision tool. Since adequate results were obtained in terms of selectivity, precision, trueness and accuracy (tolerance limits of life expectancy: ?6.0% and 3.8%) for levels of interest concentration, the methods have been considered for routine use on several samples from different provenances and collected in 4 major DRC cities. Out of 278 samples collected, 200 were eligible for analysis from which 28% were found under standards with several figures: pH failure, out of specification for amoxicillin content, absence of potassium clavulanate, physical modifications of the powders. As evidenced by these findings, medicines of low-quality continue to be a major public health problem requiring appropriate action to effectively address this problem.

关键词： Poor Quality Medicines Amoxicillin LC Method Validation Survey Study

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Peroxidase Inhibition and Antioxidant Activity of Bulk-Marketed Black Tea (Camellia sinensisL.) from the Democratic Republic of the Congo

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Journal of Biosciences and Medicines 2019年第9期7卷 66-80页

作者： Nadege Ngombe Kabamba Joyce Mbuyi Katalayi Jimmy Kabeya Kabengele Grace Nkakapen Djoumbissi Marie Joe Kabongo Kapinga Thé ophile Mbemba Fundu Dieudonné Tshitenge Tshitenge Pascal dibungi T. Kalenda Paulin Mutwale Kapepula Centre d’Etudes des Substances Naturelles d’Origine Vé gé tale (CESNOV) Faculty of Pharmaceutical Sciences University of Kinshasa Kinshasa The Democratic Republic of the Congo Laboratoire d’Analyses et de Recherche en Alimentation et Nutrition (LARAN) Department of Biology Faculty of Sciences University of Kinshasa Kinshasa The Democratic Republic of the Congo

The widely known leaves of Camellia sinensis (L.) derive the most consumed beverage globally. All over the world, like in the Democratic Republic of the Congo (DR Congo), they are marketed in the form of packaged and in bulk black tea. The aim of this study was to report the microscopic botanical features, the chromatographic fingerprints, the antioxidant activities, and the peroxidase inhibition of bulk-marketed black teas consumed in DR Congo. The microscopic analysis of powders from unpackaged black tea marketed in DR Congo allowed the identification of specific microscopical features, which are characteristic to C. sinensis (L.) such as numerous asterosclerites, numerous fragments of unicellular trichomes;fragments of abaxial epidermis consisting of cells with slightly wavy walls, numerous anomocytic stomata, and the detection of biological contaminants, e.g. the dust mites. TLC and HPLC analysis indicated that flavonoids (hyperoside, rutin) and phenolic acids (chlorogenic acid, gallic acid) were major phenolic compounds, along with other secondary metabolites. Aqueous extracts from all bulk-marketed black teas exhibited good antioxidant activities at the concentrations range of 1 - 20 μg·mL-1 using in vitro ABTS and DPPH assays. These extracts showed equally inhibitory effects on myeloperoxidase (MPO) and horseradish peroxidase (HRP) activities at the concentration ranges of 1 - 20 μg·mL-1 and 20 - 100 μg·mL-1 respectively. Bulk-marketed black teas consumed in DR Congo possess antioxidant and anti-inflammatory properties comparable to packaged Ceylon tea largely marketed. The observed bioactivities support the health benefits of the consumed black tea by Congolese people, despite the storage conditions to be improved in order to preserve its nutritional and therapeutic assets.

关键词： Antioxidant Activity Black Tea Camellia sinensis HRP L-012 Mites Myeloperoxidase

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