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John Oludele Olanlokun,1 Mercy Toluwase Ekundayo,1 Oluwakemi Ebenezer,2 Neil Anthony Koorbanally,2 Olufunso Olabode Olorunsogo1 1Laboratories for Biomembrane Research and Biotechnology, Faculty of Basic Medical Sciences, College of Medicine, University of Ibadan, Ibadan, 4000, Nigeria; 2School of Chemistry and Physics, University of KwaZulu-Natal, Durban, 4006, South AfricaCorrespondence: John Oludele OlanlokunDepartment of Biochemistry, Faculty of Basic Medical Sciences, College of Medicine, University of Ibadan, Room NB 305, Ibadan, NigeriaEmail jodel72000@yahoo.comIntroduction: Resistant malaria is a fatal disease. Globimetula braunii (African Mistletoe) is traditionally used for malarial treatment but this fact has not been scientifically reported.Methods: Plasmodium berghei (NK65)-infected male Swiss mice (20± 2 g) were treated orally and once daily with 100, 200, and 400 mg/kg BW of methanol extract and its respective hexane, dichloromethane and ethyl acetate fractions for 9 days. P-alaxin was used as control drug P. berghei (ANKA)-infected mice were then treated with the most potent fraction for 5 days. Parasitemia and parasite clearance were determined by microscopy, while hematological parameters, heme, hemozoin, and mouse erythrocyte membrane stabilisation were assayed. The phytochemicals in the most potent fraction were identified using gas chromatography-mass spectrometry.Results: Hexane fraction (HF)-treated mice (400 mg/kg BW) had the least mean parasite load (0.00 ± 0.00; 0.14 ± 0.05%) and highest clearance (100 ± 0.00; 75.50 ± 4.95%) compared with infected control (9.81 ± 0.09; 6.84 ± 0.09%) in susceptible and resistant models, respectively. Hexane fraction modulated hematological indices, minimised erythrocyte membrane damage in heat-induced (2.18 ± 0.94%) and hypotonic solution-induced (7.93 ± 0.93%) compared to artequin (5.05 ± 2.18; 6.38 ± 0.33%) and P-alaxin (67.45 ± 5.15; 56.78 ± 1.10%) in both models of membrane stabilisation, respectively. Hexane fraction (P< 0.01) increased heme and decreased hemozoin contents. Friedelan-3-one was identified as the most abundant triterpene.Conclusion: The results indicated that G. braunii has anti-plasmodial properties and minimally dis-stabilised erythrocyte membrane. The major findings in this study are that n-hexane fraction of G. braunii possess excellent and moderate antiplasmodial activity against susceptible and resistant P. berghei, respectively. This was reflected via decreased parasite load, improved hematological parameters, increased heme and decreased hemozoin contents. Friedelan-3-one, a major constituent of the n-hexane fraction, may be responsible for this activity.Keywords: Globimetula braunii, erythrocytes, heme, hemozoin, Plasmodium berghei, resistant malaria

John Oludele Olanlokun,1 Adeola Oluwakemi Olowofolahan,1 Olusola Bodede,2 Adekunle Theophilus Adegbuyi,3 Gerhard Prinsloo,2 Paul Steenkamp,4 Olufunso Olabode Olorunsogo1 1Laboratories for Biomembrane Research and Biotechnology, Department of Biochemistry, College of Medicine, University of Ibadan, Ibadan, Nigeria; 2Department of Agriculture and Animal Health, University of South Africa, Florida Campus, Florida, 1710, South Africa; 3Department of Pharmacology and Therapeutics, Faculty of Pharmacy, Federal University, Oye-Ekiti, Nigeria; 4Research Centre for Plant Metabolomics, Department of Biochemistry, University of Johannesburg, Johannesburg, 2006, South AfricaCorrespondence: John Oludele OlanlokunLaboratories for Biomembrane Research and Biotechnology, Department of Biochemistry, Faculty of Basic Medical Sciences, College of Medicine, University of Ibadan, Room NB 305, Ibadan, NigeriaTel +2348038014201Email jodel72000@yahoo.com; jo.olanlokun@mail.ui.edu.ngBackground: Inflammation is a protective response of the host to infections and tissue damage and medicinal plants have been used to regulate inflammatory response. The phytochemical contents of the n-hexane fraction of Alstonia boonei and their anti-inflammatory potentials in lipopolysaccharide-induced inflammation were investigated in rat liver.Materials and Methods: A quantity of 5 mg/kg lipopolysaccharide (LPS) was used to induce inflammation in twenty-five male Wistar rats, grouped (n = 5) and treated as follows: negative control (10 mL/kg saline), positive control (1 mg/kg ibuprofen); 50, 100 and 20 mg/kg of the n-hexane fraction of Alstonia boonei were administered to test groups. In another experiment, twenty rats (n = 5, without LPS) were administered the same doses of the n-hexane fraction of A. boonei and ibuprofen for seven days. At the end of the experiment, animals were sacrificed, serum was obtained from blood and liver mitochondria isolated in a refrigerated centrifuge. Mitochondrial permeability transition (mPT) pore opening and mitochondrial F0F1 ATPase (mATPase) were determined spectrophotometrically. Serum interleukins 1β, 6 (IL-1β, IL-6), tumour necrosis factor alpha (TNF-α), C-reactive protein (CRP) and creatine kinase (CK), gamma glutamyl transferase (GGT), aspartate and alanine aminotransferases (AST and ALT,) of the animals in which inflammation was induced using LPS but treated with graded doses of n-hexane fraction of A. boonei were determined using the ELISA technique. The phytochemical contents of the n-hexane fraction of A. boonei were determined using ultra performance liquid chromatography-tandem mass spectrometer (UHPLC-MS).Results: Calcium induced mPT in 8 fold and LPS induced mPT 14 fold in the negative control while the n-hexane fraction reversed mPT in the treated groups (50, 100 and 200 mg/kg) to 2, 4, 4 folds, respectively. LPS treatment of the negative group enhanced F0F1 mATPase activity, increased CRP, TNF-α, IL-1β, IL-6 levels as well as CK, AST, ALT and GGT activities. These values were significantly reduced by 100 and 200 mg/kg of the n-hexane fraction. UHPLC-MS analysis of the fraction revealed the presence of terpenoids, phenolics and sphingolipids.Conclusion: These results showed that bioactive phytochemicals present in the n-hexane fraction of A. boonei were not toxic, have an anti-inflammatory effect and could be used for the treatment of inflammatory diseases.Keywords: Alstonia boonei, endotoxins, inflammatory cytokines, liquid chromatography-mass spectrometry

Azadirachta indica A. Juss (Meliaceae) (AI) and Curcuma longa L. (Zingiberaceae) (CL) are used for malaria treatment but their anti-glycolytic and host mitochondrial effects have not been studied. The AI stem-bark and CL rhizomes were extracted with methanol. Methanol extract of CL (Turmeric) was partitioned to yield methanol fraction (MF). Swiss mice infected with Plasmodium berghei (NK 65 strain) were treated with 200 and 400 mg/kg of AI and turmeric for seven days. Turmeric and MF (200 and 400 mg/kg) were combined with 400 mg/kg AI to treat mice infected with Plasmodium berghe i (ANKA strain) for four days. Drug and infected controls mice were treated with artemether lumefantrine (10 mg/kg) and distilled water (10 mL/kg), respectively. Serum lactate dehydrogenase (LDH) and aldolase activities were determined. Liver mitochondria were obtained for mitochondrial permeability transition (mPT) pore opening and F o F 1 ATPase assays. The curcumin content of turmeric was determined using HPLC while LD 50 of Turmeric and AI was also determined. The AI, and its combination with turmeric decreased parasite load and increased chemosuppression in both sensitive and resistant studies while MF and its combinations with AI induced mPT pore opening. In the resistant experiment, AI + Turmeric 400 mg/kg decreased F o F 1 ATPase, LDH and aldolase activities against the infected control. The LD 50 values of both extracts were above 2000 mg/kg while the MF had the highest curcumin content. Antiplasmodial mechanisms of action of AI, CL and their combinations involve anti-glycolytic effects. Their composite formulations are more potent in malaria treatment., Competing Interests: Conflict of interestThe authors declare that there is no conflict of statement., (© Indian Society for Parasitology 2024. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

Ethnopharmacological Relevance: Andrographis paniculata (AP) ((Burm f.) Wall. ex Nees) is a medicinal plant, documented for its folkloric use in the treatment of malaria., Aim: This study was designed to determine the potency of extract and fractions of A. paniculata (AP) as a curative, both for susceptible and resistant malaria and to also determine the plant's mechanism of action. This study was also designed to determine whether AP extract and its most potent fraction will mitigate infection-mediated mitochondrial dysfunction, and to assess the phytochemical constituents of the most potent fraction., Materials and Methods: n-Hexane, dichloromethane, ethylacetate and methanol were used to partition the methanol extract of A. paniculata. Graded doses of these extract and fractions were used to treat mice infected with chloroquine-sensitive strain of P. berghei in a curative model. The most potent fraction was used to treat mice infected with resistant (ANKA strain) P. berghei. Inhibition of hemozoin formation, reversal of mitochondrial dysfunction and antiinflammatory potentials were determined. A combination of ultraperformance liquid chromatography-quadrupole time of flight-mass spectrometry and nuclear magnetic resonance spectroscopy were used for chemical analysis., Results: Microscopy revealed that the dichloromethane fraction decreased the parasite burden the most, and inhibition of the hemozoin formation is one of its mechanisms of action. The dichloromethane fraction reversed parasite-induced mitochondrial pore opening in the host, enzyme-dependent ATP hydrolysis and peroxidation of host mitochondrial membrane phospholipids as well as its antiinflammatory potentials. The UPLC-qTOF-MS report and NMR fingerprints of the dichloromethane fraction of A. paniculata yielded fourteen compounds of which sibiricinone C was identified from the plant for the first time., Conclusion: Fractions of A. paniculata possess antiplasmodial effects with the dichloromethane fraction having the highest potency. The potent effect of this fraction may be attributed to the phytochemicals present because it contains terpenes implicated with antimalarial and antiinflammatory activities., Competing Interests: Declaration of competing interest The authors declare that they have no competing financial interests., (Copyright © 2024. Published by Elsevier B.V.)

Polycystic ovary syndrome (PCOS) is a gynecological disorder among reproductive-aged women and a major cause of infertility. Different treatment options are being employed but with side effects. This has mandated alternative treatment options, especially complementary therapy. This study therefore investigated the possible protective effects of methanol extract of Drymaria cordata in Letrozole-induced PCOS. The plant is folklorically used in the treatment of diverse ailments including PCOS, fibroids, uterine/ovarian/breast tumors, and cancers. Forty-eight female Wistar rats were acclimatized and initially divided into two groups: group I(control group) and group II(PCOS group). PCOS was induced by the oral administration of letrozole/high-fat diet for 21 days. After the induction, the PCOS group was sub-divided into four groups (n = 4): group II (positive control with PCOS), group III (MET 2mg/kg), group IV (MEDC 200mg/kg), and group V (MEDC 400mg/kg). Rats were orally treated with MET and MEDC for six weeks after the PCOS induction. At the end of the experimental period, blood samples were collected, sera were separated, mitochondria were isolated, and the mPT, some apoptotic biomarkers, hormonal and lipid profiles, and oxidative stress markers were determined. Ovarian histological evaluation and GC-MS analysis of MEDC were carried out. There was no significant mPT pore opening in the PCOS (untreated) group. However, treatments with MEDC caused significant mPT pore opening, upraised caspase 9, caspase 3, and Bax, and decreased anti-apoptotic Bcl-2 levels. The MEDC treatments restored the hormonal and lipid profiles, increased the levels of GSH-Px and SOD and decreased TBARS. Histological examination revealed resolved ovarian cysts and improved follicular growth with MEDC treatments. Comparable results were observed for both MEDC and metformin. The GC-MS analysis revealed the presence of some major pharmacologically relevant compounds. These findings suggest that MEDC contains phytochemicals that can protect against letrozole-induced PCOS possibly by normalizing the impaired hormonal balance, restoring the lipid profile, and improving the antioxidant milieu of the system., (© 2024. The Author(s), under exclusive licence to Society for Reproductive Investigation.)

Plasmodium infection is a health challenge. Although, antiplasmodial drugs kill the parasites, information on the effects of infection and drugs on the expression of some genes is limited. Malaria was induced in two different studies using NK65 (chloroquine-susceptible, study 1), and ANKA (chloroquine-resistant, study 2) strains of Plasmodium berghei in 30 male Swiss mice (n = 5) in each study. Mice orally received 10 mL/kg distilled water, (infected control), Mefloquine (MF) (10 mg/kg), MF and Curcumin (CM) (25 mg/kg), MF and CM (50 mg/kg), CM (25 mg/kg) and CM (50 mg/kg). Five mice (un-infected) were used as the control. After treatment, total Ribonucleic acid (RNA) was isolated from liver and erythrocytes while Deoxyribonucleic acid (DNA)-free RNA were converted to cDNA. Polymerase Chain Reaction (PCR) amplification was performed and relative expressions of FIKK12, AQP3, P38 MAPK, NADH oxidoreductase, and cytochrome oxidase expressions were determined. Markers of glycolysis, toxicity and antioxidants were determined using ELISA assays. While the expression of FIKK12 was blunted by MF in the susceptible study, co-treatment with curcumin (25 mg/kg) yielded the same results in the chloroquine-resistant study. Similar results were obtained on AQP3 in both studies. Curcumin decreased P38 MAPK in both studies. Plasmodium infection decreased NADH oxidoreductase and cytochrome oxidase but mefloquine-curcumin restored the expression of these genes. While glycolysis and toxicity were inhibited, antioxidant systems improved in the treated groups. Curcumin is needed for effective therapeutic efficacy and prevention of toxicity. Plasmodium infection and treatment modulate the expressions of some genes in the host. Curcumin combination with mefloquine modulates the expression of some genes in the host., Competing Interests: Authors declare that no competing interest exists., (© 2024 Published by Elsevier B.V.)

Introduction: Unmanaged Diabetes Mellitus (DM) usually results to tissue wastage because of mitochondrial dysfunction. Adverse effects of some drugs used in the management of DM necessitates the search for alternative therapy from plant origin with less or no side effects. Ocimum gratissimum (L.) (OG) has been folklorically used in the management of DM. However, the mechanism used by this plant is not fully understood. This study was designed to investigate the effects of chloroform fraction of OG leaf (CFOG) in the reversal of tissue wastage in DM via inhibition of mitochondrial-mediated cell death in streptozotocin (STZ)-induced diabetic male Wistar rats. Methods: Air-dried OG leaves were extracted with methanol and partitioned successively between n -hexane, chloroform, ethylacetate and methanol to obtain their fractions while CFOG was further used because of its activity. Diabetes was induced in fifteen male Wistar rats, previously fed with high fat diet (28 days), via a single intraperitoneal administration of STZ (35 mg/kg). Diabetes was confirmed after 72 h. Another five fed rats were used as the normal control, treated with corn oil (group 1). The diabetic animals were grouped (n = 5) and treated for 28 days as follows: group 2 (diabetic control: DC) received corn oil (10 mL/kg), groups 3 and 4 were administered 400 mg/kg CFOG and 5 mg/kg glibenclamide, respectively. Body weight and Fasting Blood Glucose (FBG) were determined while Homeostasis Model Assessment of Insulin Resistance (HOMA-IR) and beta cell (HOMA-β), and pancreatic tissue regenerating potential by CFOG were assessed. Activity-guided purification and characterization of the most active principle in CFOG was done using chromatographic and NMR techniques. The animals were sacrificed after 28 days, blood samples were collected and serum was obtained. Liver mitochondria were isolated and mitochondrial permeability transition (mPT) was investigated by spectrophotometry. Results: CFOG reversed diabetic-induced mPT pore opening, inhibited ATPase activity and lipid peroxidation. CFOG reduced HOMA-IR but enhanced HOMA-β and caused regeneration of pancreatic cells relative to DC. Lupanol was a major metabolite of CFOG. Discussion: Normoglycemic effect of CFOG, coupled with reversal of mPT, reduced HOMA-IR and improved HOMA-β showed the probable antidiabetic mechanism and tissue regenerating potentials of OG., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Salemcity, Olanlokun, Olowofolahan, Olojo, Adegoke and Olorunsogo.)

A recent review on the ethnomedicinal, chemical, pharmacological, and toxicological properties of Alstonia boonei revealed the plant's potential in the treatment and management of a range of diseases. However, most of these pharmacological effects are only traceable to the crude form of the plant extract and not specific natural products. Phytochemical investigation of the methanol fraction of the methanol extract of the stem-bark of Alstonia boonei led to the isolation and identification of 2-methyl-3-propylbutane-1,4-diol. The structures were elucidated by the application of 1D-, and 2D-NMR spectroscopic analyses and by comparison with literature data. In this study, the membrane stabilizing activity, mitochondrial membrane permeability transition pore opening, cytochrome c release, mitochondrial ATPase activity, and prevention of mitochondrial lipid peroxidation activity of 2-methyl-3-propylbutane-1,4-diol (MPBD) isolated from A. boonei were determined. The results showed that MPBD significantly (p < .05) prevented peroxidation of mitochondrial membrane lipids and hemolysis using both the heat-induced and hypotonic solution-induced membrane stabilization assays. On the contrary, the compound caused large amplitude swelling of rat liver mitochondria in the absence of calcium, significant (p < .05) cytochrome c release and enhancement of mitochondrial ATPase activity in vitro. Our findings suggest that MPBD showed characteristic biological properties useful in modulating cell death., (© 2022 John Wiley & Sons Ltd.)

Background: The diagnosis of uterine dysfunction (endometrial hyperplasia) is on the rise. The available treatment is quite expensive and associated with some side effects. The therapeutic potential of natural products is now being explored, as they are easily available with little or no side effects. Drymaraia cordata is folklorically utilized in the treatment of diverse ailments including uterine fibroids., Objectives: This study aims to investigate the potential therapeutic effect of chloroform fraction of methanol extract of Drymaria cordata (CFDC) in estradiol benzoate (EB)-induced endometrial hyperplasia., Methods: Thirty-six rats were randomly divided equally into six groups. These included control group, CFDC: (100 mg/kg), CFDC: (200 mg/kg), EB: (2 mg/kg), EB + CFDC (100 mg/kg), and EB + CFDC (200 mg/kg). Endometrial hyperplasia (EH) was induced by intraperitoneal injection of EB. The levels of estrogen (E2), progesterone (PG), Follicle stimulating hormone (FSH), Luteinizing hormone (LH), Malondialdehyde (MDA), Superoxide dismutase (SOD), and Glutathione peroxidase (GSH-Px) activities were determined using ELISA technique. The uterine histological assessment and immunohistochemical expression levels of estrogen receptor, Ki-67, cytochrome c, and caspase 3 were carried out., Results: EH was severely expressed in the uterine section of EB-treated rats. However, CFDC administration improved the pathological features of the animal model. The sex hormones levels were increased in the EB-treated group, which were significantly reduced by CFDC. The antioxidant indices were also restored by CFDC. Immunoexpression levels of ERα and Ki-67 were downregulated while cytochrome c and caspase 3 were upregulated by CFDC., Conclusion: This study suggests that CFDC contains phytochemicals that can protect against EB-induced EH via modulation of hormonal signaling, apoptotic machinery, and oxidative indices., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

The present study investigated the antioxidant and cyto-/mito-protective roles of Methanol Fraction of Ficus mucoso (MFFM) in iron-induced oxidative damage in Drosophila melanogaster. At first, 10-day survival rates were carried out separately on FeSO 4 and MFFM, respectively, after which ameliorative effects of MFFM were investigated on FeSO 4 -induced toxicity for 5 days using biochemical and behavioral markers. Additionally, mitochondria were isolated from treated D. melanogaster to assess mitochondrial Permeability Transition (mPT) pore opening. The results showed that FeSO 4 significantly reduced survival rate, total thiol level and activities of catalase and glutathione-S-transferase in D. melanogaster . In addition, treatment with FeSO 4 caused increased generation of H 2 O 2 , NO (nitrite/nitrates) and acetylcholinesterase (AChE) activity compared with control ( p  < 0.05). Conversely, MFFM restored FeSO 4 -induced inhibition of glutathione-S-transferase and catalase activities, as well as glutathione and total thiol levels. FeSO 4 -induced elevation of AChE activity as well as H 2 O 2 and NO (nitrites/nitrates) levels were ameliorated by MFFM with improved climbing activity. Interestingly, MFFM prevented FeSO 4 -induced mitochondrial Permeability Transition (mPT) pore opening, and elevated mitochondrial ATPase activity and mitochondrial lipid peroxides generation in D. melanogaster. Taken together, our results demonstrated that iron impaired anti-stress defence capacity, altered behavioral functions, increased generation of mitochondrial malondialdehyde and activated opening of the mPT pore in D. melanogaster. Conversely, methanol fraction of F. mucoso protected against iron-induced cyto-/mito-toxic effects. F. mucoso possibly contain bioactive agents which might be useful in the management of disorders associated with oxidative stress induced by iron and or related metals.

Purpose: Researches have shown the relevance of antioxidants in the management of several diseases. In the present study, the effects of quercetin and vitamin E were investigated on the mitochondrial functions in vivo and in silico., Methods: Structures of quercetin and vitamin E were docked against mitochondrial Adenosine triphosphatase (mATPase), and cytochrome c cavity. Activity of liver mATPase and mitochondrial permeability transition pore opening were determined by spectrophotometry and activation of cytochrome c was examined by immunohistochemistry., Results: The binding energy of vitamin E (-9 Kcal/mol) in mATPase cavity compares well with glibenclamide (-9.4 Kcal/mol), while quercetin had a binding energy of -7.1 Kcal/mol. Similarly, vitamin E, quercetin were bound to cytochrome c by -6.4 and - 5.5 Kcal/mol energy, while glibenclamide had -7.0 Kcal/mol binding energy. The results showed that vitamin E was more accessible to the protoporphyrin prosthetic group in cytochrome c than quercetin. In the experimental studies, it was validated that vitamin E inhibited the uncontrolled activity of mATPase in diabetic rat liver. This was also proven and tested on the liver mitochondrial permeability transition pore opening observed in diabetic rats. Further experimental assessment of these on activation of cytochrome c showed that vitamin E reduced the extent of the activation more than quercetin and glibenclamide., Conclusion: There is a favorable protein-ligand interaction between quercetin and vitamin E in certain apoptotic proteins implicated in diabetes complications., Competing Interests: Conflict of interestThe authors declare no conflict of interest., (© The Author(s), under exclusive licence to Tehran University of Medical Sciences 2022, Springer Nature or its licensor holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

Ethnopharmacological Relevance: Liver diseases is a public health issue in sub-saharan Africa and has been reported to be the major cause of many hospital admissions. Oxidative stress, mitochondrial dysfunction and inflammation play important roles in several diseases including liver injury. Cajanus cajan is an indigenous medicinal plant useful in the traditional treatment of jaundice, inflammation and liver injury., Aim of Study: This study assessed the effects of methanol extract Cajanus cajan (MECC) on mitochondrial permeability transition (mPT) pore opening, biomarkers of oxidative stress and inflammation in CCl 4- induced liver injury in rats., Methods: Wistar albino rats (200-210g) were completely randomized into five (5) groups of six animals each. Group I (control) was given distilled water orally once daily. Animals in group II were administered CCl 4 in parafin (1:1) at a dose of 0.5 mL/kg i.p on the seventh day. Animals in groups III, IV and V were administered methanol extract of Cajanus cajan (MECC) at doses of 100, 200 mg/kg and silymarin (100 mg/kg) respectively for 7 days prior to a single intraperitoneal dose of CCl 4 . After 24 h of CCl 4 treatment, serum and liver tissues were collected. Mitochondrial permeability transition (mPT) pore opening, mitochondrial ATPase activities and biomarkers of oxidative stress were determined spectrophotometrically. Tumor necrosis factor (TNFα), NF-κB and COX-2 were determined by immunohistochemistry and the phytochemicals present in the extract were determined by GC-MS., Results: Liver enzyme (AST, ALP, ALT and γGT) activities and MDA levels were significantly decreased in rats pretreated with MECC at the dose of 100, 200 and silymarin (100 mg/kg) when compared to the rats administered CCl 4 alone (p < 0.05). GSH, GST, CAT and SOD increased and the expressions of TNFα, NF-κB and COX- 2 were also reduced when compared to the CCl 4 - treated animals. In addition, the liver histopathological analyses revealed that MECC markedly alleviated inflammatory cell infiltration, hepatic fibrosis, hepatocyte ballooning, necrosis and severe apoptosis of hepatocytes induced by CCl 4 . GC-MS analysis yielded 23 compounds including flavonoids, terpenoids and fatty acids., Conclusion: Cajanus cajan leaf extract elicited hepatoprotective action on CCl 4 -induced liver injury via inhibition of mPT pore opening, prevention of CCl 4 -induced hepatic oxidative stress and suppression of inflammatory response thus it may become useful for chemoprevention of liver injury. This supports its traditional use., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Background: Apoptosis is a common pathology in malaria and most antimalarial drugs induce apoptosis during chemotherapy. Globimetula braunii is an African mistletoe used for the treatment of malaria but its effect on mitochondria-mediated apoptosis is not known., Methods: Malarial infection was induced by the intraperitoneal injection of NK 65 strain Plasmodium berghei -infected erythrocytes into mice which were treated with graded doses (100-400 mg/kg) of methanol extract (ME), and fractions of n -hexane, dichloromethane, ethylacetate and methanol (HF, DF, EF and MF) for 9 days after the confirmation of parasitemia. Artequine (10 mg/kg) was used as control drug. The fraction with the highest antiplasmodial activity was used (same dose) to treat mice infected with chloroquine-resistant (ANKA) strain for 5 consecutive days after the confirmation of parasitemia. P-alaxin (10 mg/kg) was used as control drug. On the last day of the treatment, liver mitochondria were isolated and mitochondrial Permeability Transition (mPT) pore opening, mitochondrial F 0 F 1 ATPase (mATPase) activity, lipid peroxidation (mLPO) and liver deoxyribonucleic acid (DNA) fragmentation were assessed spectrophotometrically. Caspases 3 and 9 were determined by Enzyme-Linked Immunosorbent Assay (ELISA) technique. Cytochrome c, P53, Bcl-2-associated X protein (Bax), and B-cell lymphoma-2 (Bcl2) were determined via immunohistochemistry. Phytochemical constituents of the crude methanol extract of Globimetula braunii were determined via the Gas Chromatography-Mass Spectrometry (GC-MS) analysis., Results: There was large amplitude mPT induction by malaria parasites, extract and fractions of Globimetula braunii . At 400 mg/kg, HF significantly ( p  < 0.01) downregulated mATPase activity, and mLPO in both (susceptible and resistant) models, caused DNA fragmentation ( P  < 0.0001), induced caspases activation, P53, bax and cytochrome c release but downregulated Bcl2 in both models. The GC-MS analysis of methanol extract of Globimetula braunii showed that α-amyrin is the most abundant phytochemical., Conclusion: The n -hexane fraction of Globimetula braunii induced mitochondrial-mediated apoptosis through the opening of the mitochondrial pore, fragmentation of genomic DNA, increase in the levels of P53, bax, caspase 3 and 9 activation and cytochrome c release with concomitant decrease in the level of Bcl2. α-Amyrin is a triterpene with apoptotic effects., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2022 The Authors.)

Apoptosis is upregulated in all forms of diabetes, and the mitochondria act as target in diabetes pathophysiology. Quercetin and vitamin E have both shown usefulness in the delay of progression of diabetes-induced complications. However, their effect on the apoptotic process in diabetes mellitus is unknown. We hypothesize that quercetin treatment in diabetes may decrease the propensity for cardiomyocytic death via regulation of the mitochondria permeability transition (mPT) pore opening. Hearts from normal and streptozotocin-induced diabetic rats were used for the study. Low ionic strength heart mitochondria were used for swelling assay and mitochondrial lipid peroxidation (mLPO) activity was spectrophotometrically assessed. Levels of cytochrome c and caspase 3 and 9 were determined by immunohistochemistry, while lesions assessed by histology. Diabetic heart mPT pore showed larger amplitude swelling than control, while mLPO levels were increased in diabetic rats relative to control, this resulted in cytochrome c release. This initiated increased caspase 3 and 9 activity in diabetic rats (p < 0.05). Histology showed hemorrhagic lesions in diabetic rat hearts. Quercetin and vitamin E treatment reversed these effects, suggestive of their anti-apoptotic effect. Quercetin and vitamin E protection in diabetes is mediated by mPT pore inhibition and modulation of mitochondrial-mediated apoptosis., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Complete malarial therapy depends largely on the immunological and inflammatory response of the host to the invading potentials of malarial parasite. In this study, we evaluated the roles of betulinic acid on immunological response, anti-inflammatory potentials, cardiac and skeletal muscle tissue damage in mice infected with chloroquine susceptible (NK 65) and resistant (ANKA) strains of Plasmodium berghei . Serum Interleukins 1β and 6 (IL-1β, IL-6), tumour necrosis factor alpha (TNF-α), immunoglobulins G and M (IgG and IgM), C-reactive protein (CRP) and creatine kinase (CK) were determined using ELISA technique. Aspartate aminotransferase (AST), alanine aminotransferase (ALT) and gamma glutammyl transferase (GGT) were determined using ELISA technique. The results showed that betulinic acid decreased the levels of IL-1β, IL-6, TNF-α and CRP relative to the infected control. The IgG and IgM levels significantly increased in both models while CK did not decrease significantly in both models although serum AST, ALT and GGT significantly decreased compared to the infected control. These results showed that betulinic acid possessed anti-inflammatory, immunomodulatory and remediating effects on tissue damage. Furthermore, the decrease in activity of CK brought about by betulinic acid is indicative of decrease in cardiac and skeletal muscle injury which is a major pathological concern in Plasmodium infection and treatment., Competing Interests: Conflict of interestThe authors declare that no conflict of interest exists., (© Indian Society for Parasitology 2021.)

Pro-inflammatory signaling, cell death, and metalloproteinases activation are events in Plasmodium infection. However, it is not known if treatment with mefloquine (MF), and curcumin (CM) supplementation, will modulate these conditions. Malaria was induced in two different studies using susceptible (NK 65, study 1) and resistant (ANKA, study 2) strains of mouse malaria parasites (Plasmodium berghei) in thirty male Swiss mice (n = 5) in each study. Following confirmation of parasitemia, mice received 10 mL/kg distilled water (infected control), MF (10 mg/kg), MF and CM (25 mg/kg), MF and CM (50 mg/kg), CM (25 mg/kg) and CM (50 mg/kg). Five mice (not infected) were used as control. After treatment, the animals were sacrificed, serum obtained and liver mitochondria were isolated. Serum Tumour Necrosis Factor alpha (TNF-α), C-reactive protein (CRP), Interleukins-1 beta (IL-1β) and Interleukins-6 (IL-6) as well as caspases-3, 9 (C3 and C9), p53, serum troponin I (TI) and creatine kinase (CK), were assayed using ELISA techniques. Mitochondrial membrane permeability transition (mPT) pore opening, mitochondrial F 0 F 1 ATPase activity, and lipid peroxidation (mLPO) were determined spectrophotometrically. Matrix metalloproteinases 2 (MMP-2) and 9 (MMP-9) expressions were determined using electrophoresis. CM supplementation (25 mg/kg) significantly decreased serum p53, TNF-α, CRP and IL-6 compared with MF. In the resistant model, CM prevented mPT pore opening, significantly decreased F 0 F 1 ATPase activity and mLPO. MF activated caspase-3 while supplementation with CM significantly decreased this effect. Furthermore, MMP-2 and MMP-9 were selectively expressed in the susceptible model. Malarial treatment with mefloquine elicits different cell death responses while supplementation with curcumin decreased TI level and CK activities., (Copyright © 2021 The Authors. Published by Elsevier Masson SAS.. All rights reserved.)

Ficus mucoso is traditionally used to treat bronchial infections. This study compared the efficacy of terpene-rich fractions of F. mucoso root bark on lipopolysaccharide(LPS)-induced inflammation, liver mitochondrial permeability transition (mPT), an index of mitochondrial health, and associated pathological alterations. Terpene-Rich Fractions of Dichloromethane (TRDF) and Ethylacetate Fractions of F. mucoso (TREF) were obtained according to standard procedures. To induce systemic inflammation, a single intraperitoneal injection of 1mgLPS/kgbw was given to mice. Spectrophotometric techniques were used to evaluate the effects of the oral administration of TRDF and TREF (3 days) on levels of pro-inflammatory mediators (TNF-α, IL-1β, IL-6) using ELSA techniques as well as antioxidant indices in normal and LPS-treated mice. The mPT pore opening, mitochondrial ATPase activity and lipid peroxidation were monitored spectrophotometrically. Our results revealed that treatment with LPS caused significant elevation in serum cytokine levels while administration of 50 and 100 mg/kg TRDF and TREF significantly reduced elevated serum levels of cytokines (TNF-α, IL-1β, IL-6) in LPS-challenged mice. In addition, activitities of superoxide dismutase, catalase and liver marker enzymes (ALT and AST) as well as levels of mitochondrial lipid peroxides were significantly reduced in mice treated with TRDF and TREF relative to LPS-fed mice. Furthermore, LPS caused induction of opening of the liver mPT pore which was significantly inhibited by TRDF at 100 and 200 mg/kg bw by 71% and 88%, respectively, but only at 100 mg/kg TREF. Furthermore, mitochondrial ATPase activity was inhibited largely by TRDF. UPLC-ESI-MS analysis revealed the presence of terpenoid derivatives and a few aromatic metabolites in TRDF. The terpene dominance of TRDF metabolites was further justified on the 1 H NMR fingerprint. Overall, TRDF is more effective as a cocktail of anti-inflammatory compounds than TREF against LPS-induced acute systemic inflammation., (© 2021. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

The role of quercetin and vitamin E treatment against streptozotocin (STZ)-induced testicular abnormalities in diabetic rats and the possible mechanism of action they use for protection were investigated. Diabetes was induced by STZ (45 mg/kg i.p. once) and blood glucose was determined. Plasmatic insulin, testosterone, luteinising hormone and follicle-stimulating hormone (FSH) were determined by ELISA. Levels of cytochrome c, caspase 3 and caspase 9 were evaluated by immunohistochemistry, while lesions were viewed by histology. Insulin played a role in testicular protection against male infertility through modulation of luteinising hormone (LH). This consequently increased Leydig and Sertoli cells and maturation of germ cells with the attached epididymis having abundant spermatozoa. The study showed a positive correlation in the levels of LH, FSH and testosterone; it was further established that all treatments normalised diabetes-induced alterations. Treatment with quercetin and vitamin E resulted in 34% decrease of apoptogenic cytochrome c release. This protected the testes against excessive apoptosis by decreasing caspase 3 and caspase 9 activation by up to 30 and 28% respectively (p < .05). Histology also showed that treatment prevented testicular cell death. The findings show that quercetin/vitamin E possess free radical scavenging properties that protected against testicular damage in diabetes. This suggests the possibility of pharmaco-therapeutic intervention., (© 2021 Wiley-VCH GmbH.)

Different aspects of reproductive functions are regulated by mitochondrial-controlled events. This study investigated the effect of plumbagin (PL) on testicular mitochondria with a view to unravelling the mechanism of the antifertility potential of plumbagin in testis of healthy rats. Thirty-two male Wistar strain albino rats were randomly allocated into four groups of eight animals each. The control or healthy group received orally 0.1 % DMSO while animals in the remaining three groups received 2.5 mg PL/kg bdwt, 5.0 mg PL/kg bdwt and 10 mg PL/kg bdwt, respectively, for 14 days. In study two, twenty-four male Wistar rats were randomly divided into three (3) groups and were orally administered 0.1% DMSO (control), 30 and 100 mg/kg PL, respectively once daily for 72 h. Rat testis mitochondria were isolated using differential centrifugation. The mitochondrial Permeability Transition (mPT) pore, mitochondrial ATPase (mATPase) activity and mitochondrial lipid peroxidation were assessed spectrophotometrically. Expression of apoptotic proteins (p53, Bax, Bcl-2) and the release of cytochrome c were determined by immunochemical technique. Reproductive receptors (FSH, PR), the expression of aromatase, Testis Specific Kinase-1 {TESK-1} were quantified by RT-PCR. The various doses of plumbagin (2.5, 5.0 and 10 mg/kg bdwt) induced opening of the testicular mPT pore by 2, 5 and 8 folds, respectively, after 14 days of oral administration. These doses of plumbagin also caused enhancement of mATPase activity, elevated generation of mLPO as well as increases in the concentrations of caspases 9 and 3. Sperm analysis revealed that these doses of PL also caused significant decreases in sperm count and motility and increased sperm abnormalities compared to control. Interestingly, these effects were accompanied by dose-dependent expressions of the Bak, p53 and cytochrome c release. Conversely, the abundance of anti-apoptotic Bcl-2 protein decreased relative to control. The levels of transcripts of FSH and progesterone receptors as well as TESK-1 and aromatase decreased significantly relative to control. Furthermore, PL strongly inhibited p53-MDM2 compared to control. Altogether, these findings show that plumbagin damages testicular cells through the activation of mitochondrial pathway involving the p53 protein network., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Ethnopharmacological Relevance: Several pathological disorders have been attributed to either oxidative stress or defect in apoptotic signaling pathway. Some bioactive compounds elicit their antiproliferative properties by induction of apoptosis via mitochondrial permeability transition (mPT) pore opening., Aim of Study: The present study therefore investigated the effects of various fractions of methanol extract of Ageratum conyzoides L. (MEAC) on mitochondrial-mediated apoptosis and the possible protective potential of the most potent against monosodium glutamate (MSG)-induced hepatic damage and uterine pathological disorder. The plant is folklorically used in the treatment of cancer and gynecological disorder., Materials and Methods: The MEAC was partitioned in succession and concentrated at 40 °C to obtain chloroform(CFAC), ethylacetate(EFAC) and methanol(MFAC) fractions. Mitochondria were isolated by differential centrifugation. The opening of mPT pore, mATPase activity and hepatic DNA fragmentation were assessed spectrophotometrically. Caspases 9 and 3, SOD and GSH-Px activities and MDA level were determined using ELISA technique. Histological assessment of the liver and uterine sections and GC-MS analysis of the most potent fraction were carried out., Results: The investigation showed that oral administration of the fractions caused induction of mPT pore opening, enhanced mATPase activity, upregulated the activities of caspases 9 and 3 and also, caused hepatic DNA fragmentation with CFAC being the most potent. The CFAC reversed severe MSG-induced hepatic damage and uterine hyperplasia. The MSG-induced oxidative stress was normalized by CFAC. The GC-MS analysis of CFAC revealed the presence of some pharmacologically relevant phytochemicals., Conclusion: These findings therefore suggest that fractions of Ageratum conyzoides induce mitochondrial-mediated apoptosis. Moreover, CFAC, which is the most potent has a promising antioxidant and antiproliferative potential against MSG-induced hepatic and uterine pathological disorder., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Ethnopharmacological Relevance: Bitter melon, Momordica charantia L. (MC), is an ethnomedicinal plant cultivated in different climes. It's cytotoxic effect on several cancer cell lines has been evaluated. However, there have been contrasting reports on the actual mechanism (s) involved in the observed cell death induced by MC., Aims of the Study: To probe the mechanism of cell death induction in MDA-MB-436 (Breast) and A549 (lung) cancer cell lines treated with fractions (ethyl acetate, dichloromethane and hexane) derived from the aqueous extract of MC., Materials and Methods: Aqeous extract of the leaves of MC were fractionated using solvents of different polarities (ethyl acetate (D3), n-hexane (D4), dichloromethane (D5)). The cells were incubated with 100 and 125 μg/mL of the fractions 24 hours. Combination of fluorescence microscopy, enzyme assays, Western blot analyses and flow cytometry were employed in the study., Results: Treatment of the cells with MC fractions reduced Mitochondrial Membrane Potential (MMP) and intracellular ATP levels, while increasing reactive oxygen species levels without classical biochemical and morphological apoptotic features were seen. However, the fractions failed in upregulating either caspase-3 activation or cytochrome c release in the cancer cells., Conclusion: Overall, these results suggest that the cytotoxic effect of MC on the selected cancer cells is mediated by loss of mitochondrial function via loss of respiration leading to cell death rather than by the classical release of cytochrome c or caspase-3 activated apoptosis., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Background and Aim: This study was carried out to investigate antiplasmodial activities of Diospyros mespiliformis ( DM ) and Mondia whitei ( MW ) in Plasmodium berghei -infected mice., Materials and Methods: Air-dried stem of DM and root of MW were soaked in methanol, decanted and concentrated to give extracts. Parts of these extracts were partitioned successively to give dichloromethane, ethylacetate and methanol fractions. Mice (18 ± 3 g) were infected with Plasmodium berghei -infected erythrocytes from a donor mouse and were treated with the extracts, while the drug control group received 10 mg/kg body weight of artesunate and the parasitized control received the vehicle (5% v/v DMSO). Percentage parasitemia and clearance were estimated from thin films of blood smear. Hematological parameters were determined using standard methods. Both extracts were also tested on in vitro inhibition of β-hematin formation., Results Discussion and Conclusion: Results showed that DM had the least percentage parasitemia (0.67%) and highest percentage parasite clearance (84.7%) while the MW had 0.89% percentage parasitemia and 79.7% clearance at the highest dose used after the seventh day relative to untreated control. The cell free antiplasmodial activity of the fractions and extracts of both DM and MW revealed that DM significantly inhibited β-hematin formation than MW . The packed cell volume, white blood cell count, Lymphocyte, Eosinophil, Monocyte and Neutrophil significantly increased in the treated groups compared with the control. The results showed that the DM had higher antiplasmodial activity., (© 2020 Author(s).)

Some antimalarial drugs are immune-modulators that impact multiple pathways of innate immunity in malarial treatment. However, information on the immunomodulatory effects of artequine and rutin in the treatment of malaria remains elusive. Twenty-five Swiss mice (18 ± 2 g) were used for this study. Twenty were infected with Plasmodium berghei (NK65). Parasitemia was confirmed, and the animals were grouped (n = 5) as follows: Group A was not infected but treated orally with vehicle. Groups B to E were infected and treated (B) orally with vehicle (10 ml/kg), (C) with 10 mg/kg artequine, (D) with 10 mg/kg of artequine supplemented with 100 mg rutin/kg, and (D) with 10 mg/kg of artequine supplemented with 200 mg rutin/kg, for 7 days. Blood was collected for hematological, inflammatory cytokines, and immunoglobulins G and M assays. Post mitochondrial supernatant fraction was used for antioxidant assays. Rutin co-administration (200 mg/kg) significantly (P < 0.001) increased platelet and neutrophil counts (P < 0.01) but significantly (P < 0.01) decreased white blood cell count and lymphocyte relative to parasitized control. Also, it significantly (P < 0.05) decreased lipid peroxidation, xanthine oxidase, and superoxide dismutase activities but significantly (P < 0.05) increased reduced glutathione and glutathione S-transferase activity. Rutin co-administration also caused a significant (P < 0.001) increase in tumor necrosis factor-alpha, interleukin-6, and immunoglobulin M levels, while interleukin-1β and immunoglobulin G decreased significantly (P < 0.001) compared with parasitized control. These results showed that rutin co-administration with artequine improved host antioxidant status and modulated the immune and inflammatory responses., (© American Society of Parasitologists 2021.)

Objectives: Decoction of Adenopus breviflorus fruit is used in folkloric medicine for treating dysmenorrhea and gonorrhea. Phytochemicals from A. breviflorus may be potent in inducing mitochondrial-dependent apoptosis via the opening of the mitochondrial permeability transition (MPT) pore. Therefore, this study investigated the in vitro effects of stigmasterol isolated from the chloroform fraction of A. breviflorus (CFAB) and also the increasing concentration of CFAB on the opening of rat liver mitochondrial permeability transition (MPT) pore., Methods: Fractionation of CFAB on column chromatography yielded a needle-like crystal which structure was elucidated by standard spectroscopic techniques. The effects of stigmasterol and CFAB on MPT pore opening were assayed spectrophotometrically. Also, the effect of CFAB on mitochondrial ATPase (mATPase) activity and cytochrome c (Cyt c) release were determined., Results: Stigmasterol isolated from CFAB induced MPT pore opening significantly (p<0.05) when compared with the control. Similarly, CFAB significantly (p<0.05) induced MPT pore opening in rat liver mitochondria in a concentration-dependent manner in the presence and absence of the triggering agent - calcium ion. Furthermore, the increasing concentration of CFAB significantly (p<0.05) stimulated mitochondrial ATPase (mATPase) activity and Cyt c release in a concentration-dependent manner., Conclusions: The study showed that stigmasterol isolated from the chloroform fraction of A. breviflorus is a potent inducer of mitochondrial-dependent apoptosis. Also, the study further revealed that CFAB possesses potent bioactive compounds which can induce the mitochondrial-dependent apoptosis through the opening of the mitochondrial permeability transition pore, activation of mitochondrial ATPase (mATPase) activity and cytochrome c release., (© 2021 Walter de Gruyter GmbH, Berlin/Boston.)

Objectives: Uterine fibroids are benign tumors that develop in many women of reproductive age. Surgery is the main approach to treatment while other options are also associated with adverse effects. Studies have shown that certain bioactive agents present in medicinal plants elicit their anti-tumor activity by induction of mitochondrial permeability transition (mPT) opening. This research therefore aimed at investigating the effect of methanol extract of Annona muricata (MEAM) on mPT pore opening in normal and monosodium glutamate-induced uterine hyperplasia using female Wistar rats., Methods: Mitochondria, isolated from rat liver were exposed to different concentrations (20, 60, 100, 140 and 180 μg/mL) of MEAM. The mPT pore opening, cytochrome c release, mitochondrial ATPase (mATPase) activity and the percentage lipid peroxidation were assessed spectrophotometrically. Histological effects of MEAM on the liver, brain and uterus of normal and MSG-treated rats were investigated., Results: The in vitro results showed a significant induction of mPT pore opening by 2.4, 4.2 and 6.4 folds, release of cytochrome c and enhancement of mATPase activity at 100,140 and 180 μg/mL, respectively. However, oral administration of MEAM did not induce mPT pore opening, neither any significant release of cytochrome c nor enhancement of mATPase activity at all the dosages used. However, histological assay revealed the presence of MSG-induced cellular damage and uterine hyperplasia which was ameliorated by MEAM co-administration., Conclusions: These findings suggest that MEAM contains phytochemicals that can ameliorate MSG-induced damage and uterine hyperplasia in rats; however, the mechanism might not be via upregulation of mitochondrial-mediated apoptosis., (© 2021 Walter de Gruyter GmbH, Berlin/Boston.)

Ethnopharmacological Relevance: Diospyros mespiliformis Hochst. ex A. DC. and Mondia whitei (Hook.f.) Skeels are traditionally used in Africa for the treatment of malaria. However, scientific evidence to substantiate this folkloric claim and their effects on liver mitochondria during malaria treatment have not been reported., Aim of the Study: This study investigated the efficacy of D. mespiliformis and M. whitei against chloroquine-sensitive and resistant strains of malarial parasites in mice. It also investigated the toxicity and protection against cellular organelles like mitochondria., Materials and Methods: Male Swiss mice were infected with a chloroquine resistant (ANKA) strain of Plasmodium berghei and were treated via oral gavage with methanol extracts of D. mespiliformis and M. whitei reconstituted in diluted dimethylsulfoxide as vehicle (DMSO, 5% v/v) for five consecutive days. Percentage parasite load and clearance were assessed by microscopy. The infected control was treated with the vehicle. Hematological indices were assessed using standard procedures. Alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) were determined using assay kits. Hepatic mitochondria were isolated via centrifugation, and their permeability transition (mPT), ATPase (mATPase) activity and lipid peroxidation (mLPO) were determined spectroscopically. Liver tissue histology was carried out by standard laboratory procedures. Phytochemical analysis of both extracts were performed using LC-MS to identify the most prominent compounds from each of the extracts., Results: After treatment on day 5, D. mespiliformis and M. whitei at 400 mg/kg decreased mean values for: percentage parasitemia (5.0 ± 1.0, 2.0 ± 0.2), increased Packed Cell Volume (PCV) (36.0 ± 1.4, 36.0 ± 0.0%) and platelets (2.0 ± 1.4, 2.0 ± 2.8 × 10 5 mm 3 ) relative to the untreated control (20.0 ± 5.2; 30.0 ± 0.0%; 1.4 ± 1.4 × 10 5 mm 3 , respectively). At the same dose, D. mespiliformis and M. whitei decreased ALT (8.0 ± 3.8, 24.2 ± 4.0U/L), AST (6.2 ± 0.8, 8.0 ± 0.9U/L) and ALP (56.0 ± 0.7, 51.0 ± 1.0U/L) activities compared to the infected control (77.0 ± 10.9U/L, 14.0 ± 0.7U/L and 76.0 ± 6.0U/L, respectively). Both D. mespiliformis and M. whitei reversed mPT opening, decreased mATPase enhancement and mLPO, relative to the control. Histopathology of the liver showed extensive hemorrhagic lesions and severe disseminated congestion in the infected control while both D. mespiliformis and M. whitei were well tolerated at the highest dose. The LC-MS analysis of D. mespiliformis showed the presence of betulinic acid, tocopherol and kaempferol with antimalarial and antioxidant properties while the M. whitei sample contained coumarin and chlorogenic acid that have antimalarial and hepato-protective properties., Conclusions: D.mespiliformis and M. whitei show antimalarial effects against resistant Plasmodium berghei infection, enhanced cell viability, mito-protection and are not toxic in mice., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Ethnopharmacological Relevance: Hyperplasia, Tumors and cancers are various forms of proliferative disorders affecting humans. Surgery is the main treatment approach while other options are also associated with adverse effects. There is therefore a need for the development of better alternative therapy that is cost effective and readily available with little or no adverse effect. Some bioactive agents in medicinal plants exhibit their anti-proliferative potential by induction of mitochondrial permeability transition pore (mPT) opening. Gloriosa superba, a medicinal plant, is folklorically used in the treatment of tumors and cancers., Aim of the Study: This study therefore aimed at investigating the effect of ethanol leaf extract of Gloriosa superba (EEGS) on mPT and monosodium glutamate-induced proliferative disorder in some specific tissues using rat model., Materials and Methods: Isolated rat liver mitochondria were exposed to different concentrations (10, 30, 50, 70 and 90 μg/ml) of EEGS. The mPT pore opening, cytochrome c release, mitochondrial ATPase activity and lipid peroxidation were assessed spectrophotometrically. Caspases 9 and 3 activities were carried out using ELISA technique. Histological assessment of the liver, prostate and uterus of normal and monosodium glutamate (MSG)-treated rats were carried out., Results: The results showed significant induction of mPT pore opening, release of cytochrome c, enhancement of mitochondrial ATPase activity, inhibition of lipid peroxidation and activation of caspases 9 and 3 activities by EEGS. The histological assessment revealed the presence of MSG-induced hepato-cellular damage, benign prostate hyperplasia and uterine hyperplasia which were ameliorated by EEGS co-administration., Conclusions: These findings suggest that EEGS contains putative agents that can induce apoptosis via induction of mPT pore opening and as well protect against MSG-induced hepato-cellular damage and proliferative disorder in prostate and uterus., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Early detection and treatment of endometrial hyperplasia (EH) is mandatory for endometrial cancer prevention. Several bioactive agents of plant origin have been shown to elicit their chemotherapeutic effect against tumors and cancer via induction of mitochondrial permeability transition(mPT) pore opening. This research was therefore aimed at evaluating the potential chemopreventive effect of methyl palmitate (MP), on estradiol benzoate(EB)-induced EH, looking at the mitochondrial-mediated pathway and other possible mechanisms of action. Mitochondria were isolated using differential centrifugation. The mPT pore, mitochondrial ATPase (mATPase) activity, lipid peroxidation and cytochrome c release were determined by standard methods using spectrophotometer. Uterine interleukin 1b, MDA levels and SOD, GSH activities, were determined using commercially available kits. The uterine histological and immunohistochemical assessment of estrogen receptor (ERα), IL-1b and caspas-3 were carried out. The fibroblast cell count density was determined using histomorphometry. At all the concentrations of MP used, there was no significant induction of mPT pore opening, neither any enhancement of mATPase activity nor release of cytochrome c when compared to the control. Similar pattern of results were recorded for the in vivo study. However, there was marked increase in the uterine MDA and interleukin 1b levels, with concurrent decrease in SOD and GSH activities, in the EB-treated group, which was significantly reversed by MP co-administration. Endometrial Hyperplasia observed in the EB-treated group was ameliorated by MP co-administration. The immunoexpression of ERα and IL-1b in the EB-treated group was reversed by MP co-administration. This study suggests anti-inflammatory, antioxidant and anti-proliferative potential of MP against EB-induced EH.

Malaria is a parasitic disease that has defied many treatment plans. This study was carried out to investigate the host mitochondrial response to malarial infection and selected antimalarial chemotherapy using murine models. The effects of artesunate (ART) and proguanil (PRG) on mitochondrial Permeability Transition (mPT), mitochondrial ATPase (mATPase), level of malondialdehyde (MDA) and activities of antioxidant enzymes; catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), Xanthine oxidase (XO), glutathione S-transferase (GST) and reduced glutathione (GSH) were estimated in Plasmodium berghei -infected mice treated with ART and PRG. Besides, apoptotic markers, such as caspases 3, 9 and DNA fragmentation were estimated. Unparasitised (NORMAL) and parasitized but untreated (PU) animals were used as controls. The mPT pore opening fold of 9 (ART), 3 (PRG), and 4 (PU) were observed relative to calcium (23) for in vivo study. In vitro , graded concentrations (20, 40, 80 and 160 μg/mL) of ART gave mPT induction folds of 1, 21, 23 and 25, respectively, relative to calcium (9) while PRG did not have effect in the absence of calcium. In vivo , ART significantly ( p  < 0.001) enhanced mATPase activity than PRG. The PRG and ART increased the MDA levels in vivo. Oral administration of ART and PRG altered antioxidant enzymes status, Caspases 3 and 9 were significantly activated in PRG-treated groups; there was significant increase in DNA fragmentation in PU and PRG groups compared with the normal control. The results obtained showed that malaria parasite and antimalarial drugs cause mitochondrial-mediated apoptosis.

Monosodium glutamate (MSG) is a major food additive used as a flavor enhancer. A lot of controversies have been generated over the use of MSG. The present study therefore investigated whether MSG would induce cytotoxicity via the induction of mitochondrial permeability transition (mPT) pore opening. 36 male albino rats were used for this study. The rats were equally divided into six groups: group I is the control while group II, III, IV, V, and VI were orally treated with MSG (25, 50, 100, 200, and 400 mg/kg) daily for 28 days. The opening of the pore, cytochrome c release, mitochondrial ATPase activity, mitochondrial lipid peroxidation and hepatic DNA fragmentation were determined spectrophotometrically. Histological assessment of prostate and brain was carried out. The results show that MSG at concentrations ≤30 µg/ml did not induce mPT pore opening while higher concentrations caused significant induction of pore opening. Also, at lower doses (25 and 50 mg/kg), MSG did not cause any significant induction of mPT pore opening while at higher doses, there were significant induction of pore opening. Similar trend of results was recorded for cytochrome c release, mitochondrial ATPase activity and lipid peroxidation. The histological results show that at low doses (25 and 50 mg/kg), no significant lesion was observed while higher doses caused benign prostatic hyperplasia (BPH) in the prostate and necrotic damage in the brain. MSG administration at low dose is tolerable while high doses induce cytotoxicity via mPT pore opening.

Objectives: Broad spectrum antimalarial drugs without deleterious effects on mitochondria are scarce. It is in this regard that we investigated the potency of methanol extract and solvent fractions of Phyllanthus amarus on chloroquine-susceptible and resistant strains of Plasmodium berghei , toxicity and its consequential effects on mitochondrial permeability transition (mPT) pore opening., Methods: Malaria was induced in male Swiss mice with susceptible (NK 65) strain, divided into groups (n=5) and treated with 100, 200 and 400 mg/kg of methanol extract, n -hexane, dichloromethane, ethylacetate and methanol fractions daily for seven days. Percentage parasitemia and parasite clearance were determined microscopically. The two most potent fractions were tested on resistant (ANKA) strains. Heme and hemozoin contents were determined spectrophotometrically. The mPT, mitochondrial ATPase (mATPase) and lipid peroxidation (mLPO) were determined spectrophotometrically. Similar groups of animals were used for toxicity studies., Results: Dichloromethane fraction (400 mg/kg) had the highest antimalarial curative effect via least parasitemia (0.49) and high clearance (96.63) compared with the negative control (10.08, 0.00, respectively), had the highest heme and least hemozoin contents (16.23; 0.03) compared with the negative control (8.2, 0.126, respectively). Malaria infection opened the mPT, caused significant increase in mLPO and enhanced mATPase; while dichloromethane fraction reversed these conditions. Serum ALT, AST, ALP, GGT, urea and creatinine of dichloromethane fraction-treated mice decreased relative to control. No significant lesion was noticed in liver and kidney tissue sections., Conclusions: Dichloromethane fraction of Phyllanthus amarus had the highest antimalarial activity with the highest mito-protective effect and it was well tolerated without toxic effects., (© 2020 Walter de Gruyter GmbH, Berlin/Boston.)

Background: Uterine leiomyomas (fibroids), a menace of the reproductive age, is characterized by proliferation of smooth muscle cells (hyperplasia) of the uterus. Alpha Stone Decoction is a poly-herbal formulation that is used for the shrinkage and prevention of uterine fibroids in folkore medicine., Objective: We investigated the efficacy and safety of Alpha Stone Decoction (ASD), on monosodium glutamate (MSG) induced uterine hyperplasia., Materials and Methods: Twenty-eight mature virgin female rats were randomly divided into four study groups: A-control B- MSG (200 mg/kgbw), C- MSG + ASD (100 mg/kgbw) and D- ASD 100 mg/kgbw alone. The administration was carried out by as a single daily dose via intraperitoneal route for 14 days. Total protein, triglycerides, estradiol (estrogen), progesterone, and total cholesterol levels in sera were determined using appropriate kits. Uterine hyperplasia was assessed via histomorphometric method using the mitotic image plus software to compute the fibroblast cell count density while the uteri and ovaries of animals were stained with mason-tricon stain for histological examination., Results: Administration of MSG for 14 days resulted heavy deposits of collagen connective tissue within the myometrium layers of the uteri. ASD significantly (p < 0.05) reduced fibroblast cell count in MSG-treated animals and also protected against MSG-induced damage observed in the myometrium of the uteri and ovaries of the animals. Significant increases (p < 0.05) in levels of total protein; triglycerides, progesterone, cholesterol and estrogen in the MSG-treated animals were ameliorated following administration of ASD., Conclusion: These findings suggest that ASD contains bioactive agents which reversed MSG-induced uterine hyperplasia. It may therefore be useful in reducing the proliferation of fibroblast cells and managing other symptoms associated with uterine myoma., (Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.)

This study investigated the status and sensitivity of mitochondrial permeability transition (mPT) pore in testis and liver of rats exposed to doses of galactose, an acceptable model used to mimic natural aging. Male albino rats were divided into five groups of eight animals in each group for in vivo studies and administered distilled water, 50,100, 200 and 500 mg galactose/kgbdwt, respectively, for six consecutive weeks. Mitochondria were isolated from liver and testis by differential centrifugation. Mitochondrial permeability transition (mPT) was assessed as mitochondrial swelling and was monitored spectrophotometrically. Mitochondrial lipid peroxidation, ATPase activity, antioxidant enzymes, caspase activation, interleukins, and sperm functional characteristics were also assessed. Administration of galactose (50-500 mg/kg) to male Wistar albino rats had no effect whatsoever on the testicular mPT pore. However, liver mPT pore was significantly opened. Furthermore, the enhancement of mitochondrial ATPase activity and malondialdehyde generation were observed in the liver of galactose-exposed rats. Significant alterations in antioxidant enzymes were observed in post-mitochondrial fraction (PMF) of liver and testis. There were also increases in serum levels of IL-1β and 6. In addition, caspases 9 and 3 were significantly elevated in PMF of the liver with evidence of DNA fragmentation. However, there was no significant difference in levels of caspases in PMF of testis in model groups of galactose when compared with control. These results provide evidence that testis mitochondria do not readily undergo permeability transition pore upon exposure to doses of D-galactose that induce the opening of the pore in the liver.HighlightsTesticular mitochondria are less sensitive to induction of permeability transition than liver mitochondria in rats exposed to D-galactose for 6 weeks, despite the occurrence of alterations in the antioxidant defense system and generation of ROS in sperm cells as in hepatocytes.The occurrence of mitochondrial permeability transition in liver of galactose-exposed rats is consistent with malondialdehyde production, alteration in antioxidant levels, enhanced ATPase activity, caspases-9 and 3 activation, immune dysfunction, and DNA fragmentation.The study of biochemical basis of reduced sensitivity of testis to permeability transition under conditions which the liver is extremely susceptible may become useful in age associated-neurodegenerative diseases where apoptosis is upregulated and has to be properly managed to achieve downregulation.

Background 3-(Para-fluorobenzoyl)-propionic acid (3PFBPA) is one of the metabolites of haloperidol used in the treatment of psychotic disorders. 3PFBPA is an inhibitor of mitogen-activated protein kinase (MAPK), implicated in the development of uterine fibroids (UFs) and cellular proliferation. In this study, the effect of 3PFBPA on oestradiol valerate (OV)-induced uterine hyperplasia was investigated. Methods Uterine hyperplasia was induced by intraperitoneal (i.p.) injection of OV (3 mg/kg for 12 weeks). Expression of oestrogen receptor (ER) α, β-catenin and E-cadherin were investigated via immunohistochemistry. The histology and fibroblast cell count/μm2 (using histomorphometry) were carried out. Results There was a significant increase in the levels of oestrogen, progesterone and total cholesterol in the OV-treated group when compared with the control, assessed by enzyme-linked immunosorbent assay (ELISA) kits. Oestrogen and total cholesterol were markedly reduced in the OV + 3PFBPAtreated group when compared with the OV-treated group. The OV-induced overexpression of β-catenin and ER were also ameliorated by 3PFBPA. Also, the loss of E-cadherin function in the OV-treated group was restored by 3PFBPA. The histological findings and histomorphometric results revealed the presence of uterine hyperplasia in the OV-treated rats which was significantly reversed by 3PFBPA. Histological studies revealed a protective role against OV-induced uterine damage that was found after OV + 3PFBPA co-administration. Conclusion This study demonstrated that 3PFBPA ameliorates OV-induced uterine hyperplasia in the female Wistar rat model. The findings warrant further investigation of the antifibrotic effects of 3PFBPA in humans.

Objective: Extracts of Anchomanes difformis (AD) are used in folkloric medicine to treat several diseases and infections. However, their roles in mitochondrial permeability transition pore opening are not known., Materials and Methods: The viability of mitochondria isolated from Wistar rat liver used in this experiment, was assessed by monitoring their swelling amplitude in the absence of calcium and reversal of calcium-induced pore opening by spermine. The effects of methanol extract and fraction of A. difformis (MEAD and MFAD, respectively) on Mitochondrial Membrane Permeability Transition (MMPT) pore opening, ATPase activity, cytochrome c release and ferrous-induced lipid peroxidation were assessed spectrophotometrically. Phytochemical constituents of MEAD and MFAD were assessed using Gas Chromatography- Mass Spectrometry (GC-MS)., Results: The MEAD (10, 20, 40 and 80 μg/ ml) had no effect on MMPT pore opening in the absence of Ca 2+ , whereas MFAD at 80 μg/ml had a large amplitude pore opening effect. Both MEAD and MFAD reversed Ca 2+ ‌‌-induced swelling with inhibition values of 18, 21, 24, 23% (for MEAD) and 41, 36, 35, and 26% (for MFAD) at 10, 20, 40 and 80 μg/ml, respectively. MFAD significantly enhanced F 1 F 0 ATPase activity and caused cytochrome c release. Both MEAD and MFAD significantly inhibited ferrous-induced lipid peroxidation by 33.0, 64.0, 66, and 75% (for MEAD) and 24, 25, 30, and 45% (for MFAD), respectively. The GC-MS results revealed the presence of squalene as one of the major constituents of MEAD., Conclusion: These findings suggest that MFAD can be used to induce cell death via mitochondrial permeability transition in isolated rat liver. Inhibition of lipid peroxidation by MEAD and MFAD showed that the pore opening effect of the extract and fraction was not mediated via peroxidation of mitochondrial membrane lipids.

Objective: Several pathologies arise from the inappropriate opening of the mitochondrial permeability transition (mPT) pore. In this regard, inhibition of mPT pore represents a cytoprotective approach to preserve mitochondrial function for treatment of diseases characterized by excessive tissue wastage such as diabetes mellitus. The aim of this study, therefore, was to study the effects of fractions of Ficus mucoso, a medicinal plant used in the traditional treatment of diabetes, on mPT pore in normal and streptozotocin (STZ)-induced diabetic rat liver., Methods: Different solvent fractions of the crude methanol extract of F. mucoso were obtained by vacuum liquid chromatography and were tested on the mPT pore. Of all the fractions tested, methanol fraction of F. mucoso (MFFM) was the most potent and was used for in vivo studies. Diabetes mellitus was induced by a single intraperitoneal injection of 60 mg/kg STZ, while treatment lasted for 14 d. In vivo, 30 male Wistar rats were divided into five groups: A, normo-glycemic control (distilled water); B, STZ (65 mg/kg; diabetic control); C, STZ + MFFM (25 mg/kg); D, STZ + MFFM (50 mg/kg); E, STZ + glibenclamide (5 mg/kg). The mPT, mitochondrial ATPase activity, lipid peroxidation and cytochrome c release were assessed spectrophotometrically while blood glucose levels were monitored using glucometer., Results: In vitro, the solvent fractions of F. mucoso, at all concentrations tested, had no effect on the mPT pore, in the absence of calcium, with no significant release of cytochrome c. Interestingly, calcium-dependent pore opening was inhibited by all solvent fractions of F. mucoso, with the MFFM having the highest inhibitory effect of 83% at 3 mg/mL. Induction of opening of the mPT pore, significant (P < 0.001) enhancement of mitochondrial ATPase activity and elevated malondialdehyde (MDA) levels in STZ-induced diabetes were significantly (P < 0.001) reversed by MFFM and were comparable with the effects of glibenclamide, a standard antidiabetic drug. Also, treatment with MFFM at different doses significantly (P < 0.001) reduced high serum blood glucose compared to the diabetic control., Conclusion: F. mucoso could be useful in therapeutic management of diabetes mellitus given its ability to prevent excessive tissue wastage via inhibition of pore opening, and reduction in levels of MDA and serum blood glucose., (Copyright © 2019 Shanghai Changhai Hospital. Published by Elsevier B.V. All rights reserved.)

Ethnopharmacological Relevance: Apoptosis is downregulated in all forms of cancers. The mitochondrion has been implicated in the apoptotic process and, recently has been targeted in cancer therapy because of its role in cancer progression. Medicinal plants are used in the treatment of cancer, in particular, Calliandra portoricensis (CP) in the management of prostate cancer in Nigeria ethnomedicine., Aim of the Study: This study was designed to investigate the effects of CP on mitochondrial-mediated apoptosis and cell proliferation using prostate cancer cells., Materials and Methods: Prostatic LNCaP, DU-145, lung adenocarcinoma and healthy VERO cells were used to assess cell proliferation. Cell cycle analysis was evaluated by flow cytometry. Levels of pro-apoptotic Bax, anti-apoptotic Bcl-2, Cytochrome C Release (CCR) and activation of caspases 3(C3) and 9 (C9) were determined by ELISA, while mitochondrial integrity was evaluated by Fluorescent Intensity Ratio (FIR)., Results: Methanol Fraction of C. portoricensis (MFCP) inhibited proliferation of prostatic LNCaP, DU-145, lung adenocarcinoma and healthy VERO cells with IC 50 values of 2.4 ± 0.2, 3.3 ± 0.2, 3.6 ± 0.2 and 17.9 ± 1.6 µg/mL, respectively. The growth inhibition by MFCP correlated with a 3-fold decreased expression of Bcl-2 and a 4-fold increase in Bax levels at 10 µg/mL in LNCaP cells. Furthermore, MFCP caused a 3.5-fold reduction in FIR at 10 µg/mL and induced CCR by 4.2 folds at the same concentration relative to control. The MFCP-induced CCR is associated with activation of C3 and C9 at 10 µg/mL by 4.2 and 5.1 folds, respectively which prompted cancer cells to arrest at S phase. The LC-MS analysis revealed the presence of polyphenols including gallic acid and afzelechin in MFCP., Conclusion: Taken together, MFCP- induced cell death is mediated by alteration of mitochondrial integrity and cell cycle arrest. Hence, methanol fraction of C. portoricensis may be effective for cancer pharmacotherapy., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Ciprofloxacin (CIP) and Amoxycillin/Clavulanate (AC) are broad-spectrum antibiotics that are commonly administered for treatment of various bacterial infections. Studies have reported the antiproliferative and apoptotic activities of CIP in several cancer cell lines while AC has been implicated in drug-induced liver injury. We investigated the influence of CIP and AC on mitochondrial Permeability Transition (mPT) pore, ATPase activity, and cytochrome C release of normal Rat Liver Mitochondria (RLM) spectrophotometrically. In vitro, CIP and AC induced the opening of the mPT pore in a concentration-dependent manner with evidence of cytochrome C release maximally at 70 µg/ml by 13 and 10 folds, respectively. In vivo, CIP (100, 200 mg/kgbw) significantly induced mPT pore opening with induction folds of 2.4 and 2.6, respectively. However, low dose of AC (10 mg/kgbw) had no effect whatsoever on the mPT pore while higher dose (30 mg/kgbw) significantly induced pore opening by 3.4 folds. Similarly, CIP(100 mg/kgbw) and AC (30 mg/kgbw), significantly enhanced RLM ATPase activity, induced cytochrome C release and increased levels of RLM malondialdehyde generation and triggered the activation of caspases-9 and 3 in liver post-mitochondrial fraction. There were also significant (p<0.05) elevation in levels of serum aminotransferases and white blood cell count. Our results show that prolonged use of Ciprofloxacin and Amoxicillin Clavulanate could result in mitochondrial membrane breakdown via induction of opening of mPT pore leading to expulsion of cytochrome C, lipid peroxidation and decrease in energy content in healthy liver cells. These drugs should therefore be used with caution.

Diabetes mellitus is a chronic metabolic disorder characterized by rise in blood glucose levels and generation of free radicals which could induce mitochondrial membrane permeability transition (MMPT) pore opening. This study examined the in vivo action of quercetin and vitamin E on MMPT in the liver of streptozotocin-induced diabetic rats orally pre-treated with 30 mg quercetin/kg body weight (STZQ), 10 mg vitamin E/kg body weight (STZVit.E) and 0.6 mg glibenclamide/kg body weight (STZG). Male albino wistar rats were used in the study and were injected intraperitonially with streptozotocin (STZ) (45 mg/kg body weight) in citrate buffer. The degrees of serum and tissue peroxidation, alanine and aspartate aminotransferase activity were investigated. MMPT pore was assessed as mitochondrial swelling and was monitored spectrophotometrically as changes in absorbance at 540 nm under succinate-energized condition. There was significant increase in serum glucose level, degree of tissue peroxidation, alanine and aspartate aminotransferase activity, cholesterol and triglycerides values in the diabetic control rats following streptozotocin induction. All the treatment had effect on the damage caused by streptozotocin. Quercetin exhibited the highest chemopreventive activity. Quercetin and vitamin E significantly reduced the blood glucose level, degree of tissue peroxidation and alanine and aspartate amino transferase activity. In vivo rat liver MMPT pore was opened in diabetic control rats and significantly inhibited in STZQ, STZV and STZG treated groups by 72.3%, 58.5% and 87.5% respectively. The activity of quercetin and vitamin E were substantiated by histopathological evaluation. Our study suggests that quercetin is an effective therapeutic agent for preventing hepatic tissues from oxidative stress resulting from streptozotocin-induced diabetes by inhibiting apoptotic processes in their target cells., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Objective: Aloe vera is a perennial drought resisting, succulent plant belonging to the zanthorrhoeaceae family which historically has been used for a variety of medicinal purposes. This study seeks to determine the effect of varying concentrations of Aloe vera gel (50, 100, 150, 250, and 350 µg/ml) on mitochondrial permeability transition pore (MPTP) in rat liver mitochondria (RLM) (in vitro)., Methods: Fresh Aloe gel was prepared daily from the Aloe vera leaf and the effect of the gel on mitochondrial membrane permeability transition pore opening was estimated in vitro using the spectrophotometric method of Lapidus and Sokolove., Results: Varying concentrations of Aloe vera gel (50, 100, 150, 250, and 350 µg/ml) induced (insignificantly at p < 0.05) the opening of the mitochondrial permeability transition pore in a concentration dependent manner in the absence of calcium (Δ540 nm as -0.020 ± 0.008, -0.021 ± 0.009, -0.031 ± 0.013, -0.031 ± 0.014, -0.034 ± 0.014 respectively) when compared with the control (-0.016 ± 0.009). In the presence of calcium, the various concentrations of Aloe vera gel further opened the MMPT pore with the highest effect noticed at 350 µg/ml concentration., Conclusions: These findings indicate that Aloe vera gel modulates the mitochondrial pore opening by further increasing the effect of calcium. This effect is needed in situations that requires tissue wastage such as in cancer treatment.

The importance of hormone receptor status in assigning treatment and the potential use of human epidermal growth factor receptor 2 (HER2)-targeted therapy have made it beneficial for laboratories to improve detection techniques. Because interlaboratory variability in immunohistochemistry (IHC) tests may also affect studies of breast cancer subtypes in different countries, we undertook a Web-based quality improvement training and a comparative study of accuracy of immunohistochemical tests of breast cancer biomarkers between a well-established laboratory in the United States (University of Chicago) and a field laboratory in Ibadan, Nigeria. Two hundred and thirty-two breast tumor blocks were evaluated for estrogen receptors (ERs), progesterone receptors (PRs), and HER2 status at both laboratories using tissue microarray technique. Initially, concordance analysis revealed κ scores of 0.42 (moderate agreement) for ER, 0.41 (moderate agreement) for PR, and 0.39 (fair agreement) for HER2 between the 2 laboratories. Antigen retrieval techniques and scoring methods were identified as important reasons for discrepancy. Web-based conferences using Web conferencing tools such as Skype and WebEx were then held periodically to discuss IHC staining protocols and standard scoring systems and to resolve discrepant cases. After quality assurance and training, the agreement improved to 0.64 (substantial agreement) for ER, 0.60 (moderate agreement) for PR, and 0.75 (substantial agreement) for HER2. We found Web-based conferences and digital microscopy useful and cost-effective tools for quality assurance of IHC, consultation, and collaboration between distant laboratories. Quality improvement exercises in testing of tumor biomarkers will reduce misclassification in epidemiologic studies of breast cancer subtypes and provide much needed capacity building in resource-poor countries., (© 2013.)

The antioxidant properties of robustaside B and para‑hydroxyphenol isolated from Cnestis ferruginea were measured as the rate of inhibition of thiobarbituric acid reactive substance (TBARS) production in the Fe2+/ascorbate system. The modulatory effects of the compounds on mitochondrial membrane permeability transition (MMPT) were monitored spectrophotometrically as decreases in light scattering at 540 nm. The varying concentrations of robustaside B and para‑hydroxyphenol (0.05, 0.1, 0.2, 0.25, 0.5, 0.75 and 1 mM) significantly reduced (P<0.05) the amount of TBARS generated by the Fe2+/ascorbate system by 85.3, 86.4, 86.0, 86.1, 86.0, 86.0 and 86.0% and 86.7, 81.3, 81.3, 80, 80, 82.6 and 83.1%, respectively. Similarly, quercetin, a standard antioxidant, was found to induce an 80% reduction in the amount of TBARS produced. The same IC50 value of 0.025 mM was observed for robustaside B, para‑hydroxyphenol and quercetin. Pre‑incubation of varying concentrations of robustaside B (0.125, 0.2, 0.5 and 1 mM) with succinate‑energized mitochondria induced MMPT pore opening by 0, ‑33.3, ‑59.3 and ‑218.5%, compared with control mitochondria. Para‑hydroxyphenol at 0.1, 0.2, 0.25 and 0.5 mM induced MMPT pore opening in a concentration‑dependent manner up to 0.25 mM by ‑21, ‑54.4 and ‑107.0%, respectively. Quercetin at 0.05, 0.1, 0.25, 0.5, 0.75 and 1 mM also induced MMPT pore opening in the absence of calcium in a concentration‑dependent manner by 5, 3.7, ‑42.6, ‑81.5, ‑187 and ‑161.1%, respectively. The current observations confirm the antioxidant properties of robustaside B and para‑hydroxyphenol, and indicate a potential therapeutic use of the compounds for the treatment of diseases requiring the induction of cell death, including cancer.

Objective: To examine the influence or the effect of the extracts of Brysocarpus coccineus leaves on the mitochondrial membrane permeability transition (MMPT) pore opening in rats with a view to establishing if any bioactive constituent of the plant could become useful in the chemotherapy of cancer., Materials and Methods: The effects of extracts of the leaves of Brysocarpus coccineus, a medicinal plant with anti-tumour, anti-inflammatory and analgesic properties, were assessed on rat liver mitochondrial membrane permeability transition (MMPT) pore in the presence and absence of calcium in vitro and in vivo., Results: The results obtained show that calcium ions induced the opening of MMPT pore significantly (P < 0.05) in rat liver mitochondria, while spermine inhibited calcium-induced opening of pore, indicating that the mitochondria were intact ab initio. The results further revealed the inhibitory effects of different concentrations (200, 600, 1000, 1400, and 1800 microg/ml) of the various extracts of the leaves compared with spermine. Specifically, the data revealed that chloroform and ethylacetate extracts reversed calcium-induced opening of MMPT pore in a concentration-dependent manner (74%, 79%, 85%, 86%, 87%) for the chloroform extract and (36%, 37%, 59%, 71% and 83%) for the ethylacetate extract, respectively. On the contrary, pre-incubation of normal healthy mitochondria with the extracts in the absence of calcium resulted in the induction of the MMPT pore opening to varying degrees by these concentrations of the extracts. The chloroform extract induced pore opening in a concentration-dependent manner in the order 2.4, 2.4, 2.5, 2.6 and 3.0 folds while the ethylacetate extracts induced the opening of the pore by 1.1, 1.2, 1.3, 1.3 and 1.4 folds between 200-1800 microg/ml, respectively. The results obtained using rats orally exposed to various doses of methanol extract of the leaves of B. coccineus for fourteen days showed that there was significant (p < 0.05) induction of mitochondrial membrane permeability transition pore opening in the absence of calcium in a dose-dependent manner. Maximum induction of 26-fold was obtained at 200 mg/kgbwt while the least dose (50 mg/kgbwt) gave 17 fold induction., Conclusion: The ability of the extracts of B. coccineus to induce MMPT pore opening in the absence of calcium in vitro and in vivo suggest that the leaves of the plant contain certain bioactive substances capable of inducing MMPT opening either in the original form or as formed biotrans derivative with eventual release of apoptotic proteins which may lead to apoptosis. The property of the extracts could be exploited for cancer chemotherapy when increased rate of apoptosis is required.

Background: Increased attention is now directed towards the search for novel naturally occurring anticancer agents that can induce mitochondrial membrane permeability transition (MMPT) pore opening and cell death as a chemotherapeutic mechanism to combat cancer incidence., Aim: The inductive effects of partially purified fractions of leaves of Cnestis ferruginea- on rat liver MMPT pore opening was investigated., Method: De-fatted methanol extract of leaves of Cnestis ferruginea was partitioned between water, chloroform, ethylacetate, or butanol separately in succession. The extract solutions were concentrated at 40 degrees C to obtain water (WF), chloroform (CF), ethylacetate (EF) and butanol (BF) fractions. The effects of these fractions (0.2- 1.4 mg/ml) on MMPT pore opening or mitochondrial swelling in the presence and absence of calcium were evaluated The effects of these fractions on the rat liver mitochondrial F0F1-ATPase activity were also assessed., Results: Ca(2+)-induced MMPT pore opening was inhibited by 1 mg/ml each of MECF, CF, BF, WF and EF by 75.0%, 83.0%, 88.0%, 68.0%, and 71.0%, respectively and compared with the effect of spermine, a standard inhibitor. However, in the absence of Ca2+, the fractions significantly induced MMPT pore opening in intact mitochondria by 7.0, 5.7, 0.7, 4.8, 10.9 folds, respectively. In normal rat liver mitochondria, F1F0-ATPase activity was stimulated maximally by MECF, CF, EF, BF and WF by 4.7, 12.7, 1.6, 3.6 and 1.5 folds, respectively, thus indicating that the chloroform fraction is the most potent and therefore contains the active principle in the plant., Conclusion: The present study revealed that the leaves of Cnestis ferruginea contain bioactive substances that induced mitochondrial membrane permeability transition and activated the specific activity of F0F1 ATPase. Thus, suggesting strongly that these bioactive agents may serve as a useful chemotherapeutic strategy in cancer therapy.

Malaria, the most important parasitic disease afflicting man is the leading cause of mortality and morbidity in the world. Chemotherapy remains the mainstay for the treatment and prevention of the disease in the absence of an effective vaccine. The incidence of resistance of malaria parasites to chemotherapy is increasing and complicated. This study was therefore undertaken in order to evaluate the therapeutic effects of fractions of the stem bark of A. boonei on P. berghei-induced malaria using chloroquine as control. Different doses (200 mg/kg and 400 mg/kg body weight) of methanolic extract (ME), n-hexane (HF), chloroform (CF), ethylacetate(EF) and aqueous (AF) fractions of the stem bark of A. boonei were administered orally to albino mice. Five milligrammes chloroquine base per kilogramme body weight (5 mg/kg bw) was used as positive control while the negative control mice received only the vehicle (5% v/v tween 80). The results obtained showed that the 400 mg/kg bw dose was more effective with respect to the parasite clearance than the 200 mg/kg bw dose. The 400 mg/kg bw dose of ME gave 68.1% percent parasite clearance. The CF gave the highest clearance of 98.4% at 400 mg/kg bw after 7 days treatment while chloroquine at 5 mg/kg bw gave 100% parasite clearance. The order of increasing potency of the fractions (parasite clearance) was (EF 50.0% < AF 60.3% < HF 63.1%, < CF 98.4%) indicating that the active principle in the stem bark was highest in the CF. Percentage parasitemia following exposure to these fractions also decreased in all groups in the same order and was only significant (p < 0.05) in CF (0.11%) compared to the untreated control group. The ME of A. boonei also caused increase in PCV by 15.5%. Purification enhanced PCV value as the HF and CF fractions gave 19.0% and 24.5% increases, respectively. However, 31.5% increase in PCV was obtained in the albino mice treated with chloroquine. The EF and AF gave increase of 10.0% and 11.0% increase relative to the negative control treated mice. The high bioactivity of CF and HF indicate that the putative compound(s) in A. boonei are lipophillic and further purification could enhance greater activity. Further work is required to isolate the bioactive compound for a promising antimalarial drug from the chloroform fraction.

Objective: Erythrocyte membrane Ca(2+)-ATPase activity was determined in workers occupationally exposed to lead because of the prevalence of elevated blood lead in auto-mobile workers in some urban areas in Nigeria., Materials and Methods: Blood lead levels, biochemical profiles, lipid peroxidation, basal and calmodulin-stimulated Ca(2+)-ATPase activities were determined in erythrocytes of different categories of workers occupationally exposed to lead. These subjects were mainly battery chargers (BC), spray painters (SP) and auto mechanics (MC)., Results: Estimation of erythrocyte Ca(2+)-ATPase activity in the absence of calmodulin (basal activity) in test groups indicated that there were significant reductions in the pump function and this correlated very well with the levels of lead in their blood. Specifically, blood lead levels were of the order: BC (5.5 folds) > SP (4 folds) > MC, although there was no significant difference between the blood lead levels in MC (10.60 +/- 2.55 microgPb2+/dl) and CT (8.51 +/- 4.55 microgPb2+/dl). Similarly, the order of reduction in Ca(2+)-ATPase activity was BC (69.8%) > SP (52.8%) > MC (32.6%). There was significant difference in the values obtained for MC and CT, ATPase activity being lower in MC compared to CT or healthy individuals. In the presence of calmodulin, basal ATPase activity was increased by at least four fold in erythrocytes from healthy subjects (CT) while the basal activity of the enzymes in membranes of BC, SP and MC was enhanced by about one and half times the activity of the pump in membranes from CT. The levels of total serum protein and albumin increased significantly in BC, SP and MC when compared to healthy subjects (CT). C-reactive proteins (C-RP) levels were higher in BC, SP and MC in comparison to CT. The levels of MDA were high in all lead-exposed workers, BC > SP > MC relative to CT. Although, there were significant decreases in the PCV values of all the groups occupationally exposed to lead compared to values obtained for CT, cholesterol level increased significantly only in BC when compared to the other groups., Conclusion: These observations are probably due to the integrity of the plasma membrane of these workers and the ability of the heavy metal to compete with Ca2+ in the catalytic cycle and Ca2+ transport mechanism of the pump protein.