Insights into microRNA-mediated interaction and regulation of metabolites in tomato.

microRNAs direct regulation of various metabolic pathways in plants and animals. miRNAs may be useful in developing novel/elite genotypes, with enhanced metabolites and disease resistance. We examined miRNAs in tomato. In tomato, miRNAs in the carotenoid pathway have not been fully elucidated. We examined the potential role of miRNAs in biosynthesis of carotenoids, transcript profiling of miRNAs and their possible targets (genes and transcription factors) at different development stages of tomato using stem-loop PCR and RT-qPCR. We also identified miRNAs targeting key flavonoid genes, such as chalcone isomerase (CHI), and dihydroflavonol-4-reductase (DFR). Distinct expression profiles of miRNAs and their targets were found in fruits of three tomato accessions, suggesting carotenoid regulation by miRNAs at various stages of fruit development. This was also confirmed using HPLC of the carotenoids. The present study may help in understanding possible regulation of carotenoid biosynthesis. The identified miRNAs can be exploited to enhance biosynthesis of different carotenoids in plants.

Altered presence of extra cellular matrix components in murine skin cancer: Modulation by Azadirachta indica leaf extract.

BACKGROUND AND AIM: Although, the anticancer potential of Aqueous Azadirachta indica leaf extract (AAILE) has been robustly established against cutaneous squamous cell carcinoma (SCC) in mice, however, its ability in modulating tumor associated extra cellular matrix (ECM) is largely unknown. Therefore, the present study was conceived to explore changes in ECM during murine skin cancer and its chemoprevention by AAILE. EXPERIMENTAL PROCEDURE: Skin tumors were induced using a two-stage model of carcinogenesis employing topical application of 7,12-Dimethylbenz(a)anthracene (DMBA) and 12-O-tetradecanoyl phorbol-13-acetate (TPA) as carcinogen and promoter respectively. AAILE was administered orally to the animals. Male Laca mice were divided into four groups: control, AAILE, DMBA/TPA and AAILE + DMBA/TPA. RESULTS: The tumors obtained in DMBA/TPA and AAILE + DMBA/TPA groups were histologically identified as SCC. Tumor induction in these groups was accompanied by raised serum carcinoembryonic antigen (CEA) levels when compared to control counterparts. Assessment of hydroxyproline levels and histochemical staining with sirius red and trichrome stain revealed an increase in collagen in tumors of DMBA/TPA group. An increase in glycosaminoglycans (GAGs) levels was also observed in DMBA/TPA group as made evident by biochemical studies and histochemical staining using mucicarmine and alcian blue-periodic acid schiff's stain. Administration of AAILE to DMBA/TPA treated animals caused a decrease in collagen and GAG levels along with a decrease in serum CEA levels. CONCLUSION: Skin tumors exhibited altered presence of ECM components which is indicative of a modified ECM. AAILE administration antagonised tumor associated ECM alterations which may be contributing to its chemopreventive activity as reported previously.

Aloe vera modulates X-ray induced hematological and splenic tissue damage in mice.

The present study was premeditated to examine the radioprotective effects of aqueous Aloe vera gel extract against whole-body X-ray irradiation-induced hematological alterations and splenic tissue injury in mice. Healthy male balb/c mice were divided into four groups: group 1, control; group 2, A. vera (50 mg/kg body weight) administered per oral on alternate days for 30 days (15 times); group 3, X-ray exposure of 2 Gy (0.25 Gy twice a day for four consecutive days in the last week of the experimental protocol); and group 4, A. vera + X-ray. X-ray exposure caused alterations in histoarchitecture of spleen along with enhanced clastogenic damage as assessed by micronucleus formation and apoptotic index. Irradiation caused an elevation in proinflammatory cytokines like tumor necrosis factor and interleukin-6, total leucocyte counts, neutrophil counts and decreased platelet counts along with unaltered red blood cell counts and hemoglobin. Irradiation also caused an elevation in reactive oxygen species (ROS), lipid peroxidation (LPO) levels, lactate dehydrogenase activity and alterations in enzymatic and nonenzymatic antioxidant defense mechanism in plasma and spleen. However, administration of A. vera gel extract ameliorated X-ray irradiation-induced elevation in ROS/LPO levels, histopathological and clastogenic damage. It also modulated biochemical indices, inflammatory markers, and hematological parameters. These results collectively indicated that the A. vera gel extract offers protection against whole-body X-ray exposure by virtue of its antioxidant, anti-inflammatory and anti-apoptotic potential.

Efficacy of crocin and safranal as protective agents against genotoxic stress induced by gamma radiation, urethane and procarbazine in mice.

Crocin (CRO) and safranal (SAF) are bioactive constituents of saffron (dried stigma of Crocus sativus flower), an expensive spice with medicinal properties. Aqueous extract of saffron is known for its antigenotoxic effect against environmental genotoxins/carcinogens. However, there is need to identify saffron constituents responsible for this antigenotoxic effect. The aim of our investigation was to ascertain the role of CRO and SAF as inhibitors of in vivo genotoxic stress. For this purpose, Swiss albino mice were pretreated with CRO (50-mg/kg body weight (bw))/SAF (0.025- and 0.25-ml/kg bw) by gavage for 2 days. Thereafter, the pretreated mice were exposed to the genotoxic agents: (1) gamma radiation (GR; 2 Gy), (2) urethane (URE; 800 mg/kg) and (3) procarbazine (PCB; 60 mg/kg). In addition, CRO (50 mg/kg) was co-administered with the nitrosation reaction mixture of methylurea (MU; 300-mg/kg bw) + sodium nitrite (15 mg/kg) which can form N-nitroso-N-MU in the stomach. Genotoxic damage was measured by performing the bone marrow micronucleus test. Results obtained demonstrated significant reductions in the incidence of micronucleated polychromatic erythrocytes in the bone marrow of mice pretreated with CRO/SAF before exposure to the above DNA damaging agents, GR, URE and PCB. Co-administration of CRO with the nitrosation reaction mixture led to significant decrease in genotoxicity when compared to nitrosation reaction mixture alone. Histopathological studies revealed that these saffron constituents reduced testicular cell damage induced by the test genotoxins. The cell-free DNA-nicking assay using pBR322 DNA showed significant protective effects of CRO against hydroxyl radical-induced strand breaks.

Protective role of Aloe vera against X-ray induced testicular dysfunction.

The present investigation was carried out to evaluate the possible radioprotective potential of an Aloe vera extract against whole-body X-ray irradiation-induced testicular alterations in mice. Male balb/c mice were divided into four groups: control, A. vera, X-ray and A. vera pre-treated + X-ray irradiated. Histopathological examination revealed significant structural alterations in testes after X-ray exposure, which was also associated with the presence of apoptotic cells as assessed by TUNEL assay. X-ray irradiation resulted in elevation in the levels of reactive oxygen species, lipid peroxidation, a reduction in glutathione concentration and enhanced activities of antioxidant enzymes such as glutathione reductase, glutathione peroxidase, catalase, superoxide dismutase and glutathione-S-transferase. Sperm count/motility and testosterone levels were significantly decreased in the irradiated group. Irradiated animals pre-treated with A. vera extract revealed an improvement in antioxidant status, inhibition of lipid peroxides, apoptotic cell formation and enhanced testicular parameters when compared to the X-ray-exposed group. These findings suggest that A. vera extract could ameliorate X-ray-induced damage due to its free radical scavenging properties and its potential to boost cellular antioxidant defence machinery.

Modulatory effects of Azadirachta indica leaf extract on cutaneous and hepatic biochemical status during promotion phase of DMBA/TPA-induced skin tumorigenesis in mice.

The modulation in biochemical status of skin and hepatic tissue at the time point of commencement of promotion stage of skin carcinogenesis in mice and its intervention with aqueous Azadirachta indica leaf extract (AAILE) were investigated. 7,12-Dimethylbenz(a)anthracene (DMBA, 500 nmol/100 ul of acetone) was applied topically for 2 weeks (twice weekly), followed by phorbol-12-myristate-13-acetate (TPA, 1.7 nmol/100 ul) twice weekly for 6 weeks on the depilated skin of mice and AAILE was administered orally at a dose level of 300 mg/kg body wt thrice a week for 10 weeks. DMBA/TPA treatment upregulated the phase I enzymes in skin and hepatic tissue, as revealed by the increased cytochrome P450 (CYP) and cytochrome b5 (cyt b5) levels and aryl hydrocarbon hydroxylase (AHH) activity when compared to the control group and differentially modulated the activities of phase II enzymes like glutathione-s-transferase (GST), DT-diaphorase (DTD) and uridine diphosphate glucuronosyltransferase (UDP-GT). AAILE treatment decreased the DMBA/TPA-induced increase in cutaneous CYP level and enhanced the DTD and UDP-GT activities when compared with DMBA/TPA group. In the hepatic tissue of AAILE + DMBA/TPA group, an increase in UDP-GT activity was observed when compared to DMBA/TPA group. DMBA/TPA treatment did not alter the skin lipid peroxidation (LPO) level when compared to control group, however, in the animals that received AAILE treatment along with DMBA/TPA, a significant increase in LPO was observed when compared to control group. This was associated with a decrease, in cutaneous reduced glutathione (GSH) level of AAILE + DMBA/TPA group. Enhanced LPO level was observed in the hepatic tissue of DMBA/TPA and AAILE + DMBA/TPA groups when compared to control group. However, no alteration was observed in their hepatic GSH levels. The micronuclei score in hepatic tissue did not exhibit significant inter-group differences. The results of the present study suggest that apart from skin, liver may be affected during DMBA/TPA-induced skin tumorigenesis. AAILE treatment has the ability to modulate these changes potentially influencing the process of tumor formation. These findings seem to be important to carcinogenesis and its intervention with anti-cancer agents.

Azadirachta indica acts as a pro-oxidant and modulates cell cycle associated proteins during DMBA/TPA induced skin carcinogenesis in mice.

The present study was designed to determine the modulatory effect of aqueous Azadirachta indica leaf extract (AAILE) on cell cycle-associated proteins during two-stage skin carcinogenesis in mice. Considering the dual role of reactive oxygen species in cancer and its chemoprevention, the levels of lipid peroxidation (index of peroxidative damage) were also determined. Skin tumours were induced by topical application of 7,12-dimethylbenz(a)anthracene (DMBA) as a carcinogen followed by the repetitive application of 12-O-tetradecanoylphorbol-13-acetate (TPA) as a promoter. Skin tumours obtained in the DMBA/TPA group exhibited enhanced expression of proliferating cell nuclear antigen (PCNA, index of proliferation), p21 and cyclin D1, with no alterations in p53 expression in comparison to the control group. Tumours in AAILE + DMBA/TPA group exhibited low PCNA and cyclin D1 expression and enhanced expression of p53 and p21 in comparison to the DMBA/TPA group. The skin tumours obtained in the AAILE + DMBA/TPA group exhibited high lipid peroxidation levels in comparison to the tumours obtained in the DMBA/TPA group. The observations of the present study suggest that AAILE behaves as a pro-oxidant in the tumours, thereby rendering them susceptible to damage, which eventually culminates into its anti-neoplastic action. Also, cell cycle regulatory proteins may be modulated by AAILE and could affect the progression of cells through the cell cycle.

Early active mobilisation versus immobilisation after extrinsic extensor tendon repair: A prospective randomised trial.

BACKGROUND: Whether to splint the extensor tendon repairs or to mobilise them early is debatable. Recently, mobilisation has shown favourable results in a few studies. This study was aimed to compare the two favoured protocols (immobilisation vs. early active motion) in Indian population. PATIENTS AND METHODS: Between June 2005 and June 2007, patients with extensor tendon injuries in zones V-VIII were randomly distributed in two groups: Group A, early active motion; and group B, immobilisation. Their results at 8 and 12 weeks and 6 months were compared. RESULTS: Patients in early active motion group were found to have better total active motion and early return to work. This difference was statistically significant up to 12 weeks, but not at 6 months. CONCLUSION: Early active motion following extensor tendon repair hastens patients' recovery and helps patients to gain complete range of motion at earlier postoperative period. With improved grip strength, the early return to work is facilitated, though these advantages are not sustained statistically significantly over long term.

Azadirachta indica exerts chemopreventive action against murine skin cancer: studies on histopathological, ultrastructural changes and modulation of NF-kappaB, AP-1, and STAT1.

The present study reports the histopathological, ultrastructural changes and modulation of NF-kappaB, AP-1, and STAT 1 during skin carcinogenesis in LACA mice and its intervention with Azadirachta indica. Skin tumors were induced by topical application of 7,12-dimethylbenz(a)anthracene (DMBA) (500 nmol/100 microl for 2 weeks) followed by 12-O-tetradecanoylphorbol-13-acetate (TPA) (1.7 nmol/100 microl of acetone, twice weekly) as a promoter. Male LACA mice were divided into four groups: Control, DMBA/TPA, aqueous Azadirachta indica leaf extract (AAILE), and AAILE + DMBA/TPA. AAILE was administered orally at a dose level of 300 mg/kg body weight three times a week for 20 weeks. Topical application of DMBA/ TPA to the skin resulted in well-developed squamous cell carcinomas characterized by hyperproliferation, hyperkeratosis, and corrugation of the epidermis. Degenerative changes were observed in the tumors of AAILE + DMBA/TPA-treated animals. Scanning electron microscopy revealed surface disruptions and certain rounded structures on the skin tumors of DMBA/TPA-treated animals. Topographical changes were also observed in the tumors of AAILE + DMBA/TPA-treated animals, which resembled regions of degeneration. Tumors obtained in DMBA/TPA group were associated with enhanced expression of NF-kappaB and AP-1 when compared to the control counterparts. Inhibition in tumorigenesis in response to A. indica treatment was accompanied by an overexpression of STAT 1 and AP-1 and decrease in NF-kappaB expression. The results of the present study provide a basis for the chemopreventive potential of A. indica against murine skin carcinogenesis.

Chemopreventive activity of Azadirachta indica on two-stage skin carcinogenesis in murine model.

The present study reports the chemopreventive activity of aqueous Azadirachta indica leaf extract (AAILE) in a murine two-stage skin carcinogenesis model. Skin tumors were induced by topical application of 7,12-dimethylbenz(a)anthracene (DMBA) (500 nmol/100 µL for 2 weeks) followed by 12-O-tetradecanoylphorbol-13-acetate (TPA) (1.7 nmol/100 µL of acetone, twice weekly) as a promoter. Male LACA mice were divided into four groups: control, DMBA/TPA, AAILE and AAILE + DMBA/TPA. AAILE was administered orally at a dose level of 300 mg/kg body weight thrice a week for 20 weeks. 100% tumor incidence was observed in the DMBA/TPA treated animals, whereas the AAILE + DMBA treated animals exhibited a tumor incidence of 58.3% only. A significant reduction in the mean tumor burden (54.5%) and mean tumor volume (45.6%) was observed in the mice that received AAILE along with DMBA/TPA. Topical application of DMBA/TPA to the skin resulted in well-developed carcinomas associated with decreased expression of pro-apoptotic protein such as caspase 3 and enhanced expression of antiapoptotic protein such as bcl-2 when compared with the control counterparts. However, adminstration of AAILE inhibited skin carcinogenesis with induction of pro-apoptotic proteins such as bax, caspase 3, caspase 9 and inhibition of antiapoptotic proteins such as bcl-2. These results suggest that the induction of apoptosis may be one of the mechanisms underlying the chemopreventive effects of A. indica.

Antiviral and lung protective activity of a novel respiratory syncytial virus fusion inhibitor in a mouse model.

Respiratory syncytial virus (RSV) causes bronchiolitis in young children and common colds in adults. There is no licensed vaccine, and prophylactic treatment with palivizumab is very expensive and limited to high-risk infants. Ribavirin is used as an antiviral treatment in infants and immunosuppressed patients, and its use is limited due to side-effects, toxicity to the recipient and staff, and evidence of marginal clinical efficacy. Therefore, we studied the in vivo kinetics, and the antiviral and protective properties of a novel candidate for RSV disease treatment. The drug is a small molecule (TMC353121) discovered by screening for fusion inhibitory properties against RSV in a cellular infection model. The pharmacokinetics of TMC353121 was studied in BALB/c mice and antiviral effects determined by testing viral loads in lung tissue by quantitative RT-PCR and plaque assay after intranasal RSV infection. At doses of 0.25-10 mg · kg(-1), TMC353121 significantly reduced viral load, bronchoalveolar lavage cell accumulation and the severity of lung histopathological change after infection. Treatment remained effective if started within 48 h of infection, but was ineffective thereafter. Therefore, TMC353121 is a novel potent antiviral drug, in vivo reducing RSV replication and inhibiting consequential lung inflammation, with a great potential for further clinical development.

Lycopene mediated modulation of 7,12 Dimethlybenz (a) anthracene induced hepatic clastogenicity in male Balb/c mice.

The present study was designed to evaluate the modulatory effects of lycopene against 7, 12 Dimethylbenz (a) anthracene induced clastogenicity and oxidative stress in male Balb/c mice. The animals were divided into four groups; group I served as control (vehicle treated). Animals of group III and IV were administered lycopene orally at a dose of 4 mg/kg body weight for 10 weeks. Groups II and IV were administered DMBA, i.p., at a dose level of 40 mg/kg body weight, 48 hrs before the sacrifice of animals. Exposure to DMBA clearly induced hepatic cell injury as was evident by an increase in micronucleated cell score, lactate dehydrogenase and alkaline phosphatase activities, and Lipid Peroxidation levels. When the lycopene pre-treated animals were challenged with DMBA, a decrease in micronucleated cell score was observed, which was in corroboration with the observed decrease in LDH and ALP activities and LPO levels. DMBA treatment caused an increase in the oxidative stress with consequent alterations in enzymatic antioxidant defense system. Lycopene pre-treatment boosted the antioxidant defense in group IV. Thus, the antioxidant role of lycopene could be plausible in the protective action conferred by lycopene, enabling it to be used an effective natural free radical scavenger.

Subjective evaluation of intraoperative performance of DisCoVisc in complex ocular environments.

PURPOSE: To subjectively evaluate the intraoperative characteristics of DisCoVisc during phacoemulsification in complex ocular environments. PATIENTS AND METHODS: In this prospective observational study, two experienced surgeons (ARV and CZ) performed phacoemulsification on 100 consecutive patients with cataract associated with complex ocular environments. Inclusion criteria were eyes with shallow anterior chambers (anterior chamber depth (ACD) of <2.1 mm), inadequate pupillary dilation (3 mm), dense cataract, and white mature cataract. The surgeons subjectively assessed the endpoints at each phase of phacoemulsification and various behavioural aspects of the ophthalmic viscosurgical devices (OVDs) were subsequently evaluated. RESULTS: The distribution was as follows: eyes with white mature cataract (n=18), eyes with grades 4 and 5 cataract (n=56), eyes with co-existing shallow ACD <2 mm (n=24), and co-existing small pupil size <2 mm (n=18). DisCoVisc behaved like a moderately cohesive viscoelastic in 94% of the cases. Injection of viscoelastic was easy in 38 (38%) eyes and very easy in 62 (62%) eyes. Visualization after the viscoelastic injection was excellent in 74% of the eyes. During phacoemulsification, DisCoVisc was moderately dispersive at all the stages of emulsification. The bag maintenance during IOL implantation was excellent in 56% eyes; IOL implantation was easy in 26% of the eyes and difficult in 20% of the eyes. Surgeons found viscoelastic removal easy in 68% of the eyes. At the time of OVD removal, DisCoVisc behaved like both a dispersive and a cohesive viscoelastic in 96% of the eyes. CONCLUSION: DisCoVisc provides both cohesive and dispersive properties. DisCoVisc alone, even in complex ocular environments, enabled the surgeon to achieve good intraoperative performance.

Lycopene mediated modulation of 7, 12 dimethlybenz (A) anthracene induced hepatic clastogenicity in male Balb/c mice / Modulación mediada por licopeno de la clastogenicidad hepática inducida por 7,12 dimetilbenz (A) antraceno en ratones macho BALB/C

The present study was designed to evaluate the modulatory effects of lycopene against 7, 12 Dimethylbenz (a) anthracene induced clastogenicity and oxidative stress in male Balb/c mice. The animals were divided into four groups; group I served as control (vehicle treated). Animals of group III and IV were administered lycopene orally at a dose of 4 mg/kg body weight for 10 weeks. Groups II and IV were administered DMBA, i.p., at a dose level of 40mg/kg body weight, 48hrs before the sacrifice of animals. Exposure to DMBA clearly induced hepatic cell injury as was evident by an increase in micronucleated cell score, lactate dehydrogenase and alkaline phosphatase activities, and Lipid Peroxidation levels. When the lycopene pre-treated animals were challenged with DMBA, a decrease in micronucleated cell score was observed, which was in corroboration with the observed decrease in LDH and ALP activities and LPO levels. DMBA treatment caused an increase in the oxidative stress with consequent alterations in enzymatic antioxidant defense system. Lycopene pre-treatment boosted the antioxidant defense in group IV. Thus, the antioxidant role of lycopene could be plausible in the protective action conferred by lycopene, enabling it to be used an effective natural free radical scavenger (AU)

El presente estudio se diseñó para evaluar los efectos moduladores del licopeno frente a la clastogenicidad y el estrés oxidativo inducidos por 7,12 dimetilbenz(A) antraceno en los ratones macho Balb/c. Se dividió a los animales en cuatro grupos; el grupo I sirvió de control (tratado con vehículo). A los animales de los grupos III y IV se les administró licopeno por vía oral a una dosis de 4 mg/kg de peso corporal durante 10 semanas. Los grupos II y IV recibieron DMBA, i.p., a una dosis de 40 mg/kg de peso corporal, 48 horas antes de ser sacrificados. La exposición a DMBA indujo claramente una lesión de los hepatocitos como se hizo patente por el aumento de la puntuación de células micronucleadas, y las actividades de la lactato deshidrogenasa y la fosfatasa alcalina, así como por los niveles de peroxidación lipídica. Cuando a los animales pre-tratados con licopeno se les expuso a DMBA, se observó un descenso de la puntuación de células micronucleadas, lo que corroboraba el descenso observado de las actividades LDH y ALP y de los niveles de LPO. El tratamiento con DMBA produjo un aumento del estrés oxidativo con las consiguientes alteraciones del sistema de defensa antioxidante. El pre-tratamiento con licopeno aumentó notablemente la defensa antioxidante en el grupo IV. Así pues, el papel antioxidante del licopeno podría ser plausible en la acción protectora conferida por el licopeno, permitiendo usarlo como un eliminador natural eficaz de los radicales libres (AU)

Effects of Azadirachta indica on certain hematological parameters during benzo(a)pyrene induced murine forestomach tumorigenesis.

OBJECTIVES: Exposure to environmental toxicants/carcinogens, the process of carcinogenesis itself and cancer treatments lead to several secondary pathologies including hematological complications. Considering versatile pharmacological potentials of Azadirachta indica (A. indica), the present study was designed to evaluate its effects on certain hematological parameters in benzo(a)pyrene [B(a)P] induced murine forestomach tumorigenesis bioassay protocol. METHODS AND RESULTS: For tumor induction, starting from 3rd week of experimentation, female Balb/c mice received B(a)P intragastric instillations (40 mg/kg bwt) twice a week for 4 weeks. Aqueous A. indica leaf extract (AAILE) was orally administered (100 mg/kg bwt) using blunt-tipped canula on alternate days throughout experimentation. The study was continued for 24 weeks and certain hematological parameters were examined at 4 week intervals. In mice receiving only B(a)P, hemoglobin (Hb), red blood cells (RBCs), white blood cells (WBCs), lymphocytes and monocytes decreased whereas neutrophils increased when compared to controls. However, A. indica reversed these alterations as seen in mice that received AAILE along with B(a)P when compared to only B(a)P receiving mice. In only AAILE receiving mice, increased Hb, RBCs, WBCs, lymphocytes and monocytes with decreased neutrophils were observed in comparison to control. Also, changes in eosinophils and basophils upon B(a)P exposure and their modulation by AAILE was observed. CONCLUSIONS: These findings strongly suggested the favorable effects of A. indica on hematological parameters studied and their significance with respect to overall well being, process of tumorigenesis and its chemoprevention have been discussed in the current research manuscript.

[R207910 (TMC207): a new antibiotic for the treatment of tuberculosis]. / R207910 (TMC207): un nouvel antibiotique pour le traitement de la tuberculose.

A new class of antibacterials, diarylquinolines, was identified. The lead compound, R207910 (TMC207), was able to inhibit Mycobacterium tuberculosis in vitro, in mice and in patients. R207910 targets the mycobacterial ATP synthase. In vitro, it displayed potent activities against both drug-sensitive and multidrug-resistant strains of M. tuberculosis. It was also strongly active against dormant bacilli in the Wayne's dormancy culture system, hypoxia and nitric oxide models. In the murine model, when used alone, it was as active as the triple combination of rifampicin+isoniazid+pyrazinamide. When added to the previous combination or substituted for isoniazid or rifampicin, the treatment including the combinations containing R207910 led to culture conversion after 2 months of therapy. When added to the combination used to treat MDR-TB or substituted for moxifloxacin or ethionamide, the combinations containing R207910 led to culture conversion after 2 months of therapy. In MDR-TB infected patients, R207910 combined with second line drugs was able to convert more sputum cultures (47.6%) than the placebo combined to second line drugs regimen (8.7%).

Rates and mechanisms of resistance development in Mycobacterium tuberculosis to a novel diarylquinoline ATP synthase inhibitor.

R207910 (also known as TMC207) is an investigational drug currently in clinical studies for the treatment of multidrug-resistant (MDR) tuberculosis. It has a high degree of antimycobacterial activity and is equally effective against drug-susceptible and MDR Mycobacterium tuberculosis isolates. In the present study, we characterized the development of resistance to R207910 in vitro. Ninety-seven independent R207910-resistant mutants were selected from seven different clinical isolates of M. tuberculosis (three drug-susceptible and four MDR isolates) at 10x, 30x, and 100x the MIC. At a concentration of 0.3 mg/liter (10x the MIC), the mutation rates ranged from 4.7 x 10(-7) to 8.9 x 10(-9) mutations per cell per division, and at 1.0 mg/liter (30x the MIC) the mutation rate ranged from 3.9 x 10(-8) to 2.4 x 10(-9). No resistant mutants were obtained at 3 mg/liter (100x the MIC). The level of resistance ranged from 0.12 to 3.84 mg/liter for the mutants identified; these concentrations represent 4- to 128-fold increases in the MICs. For 53 of the resistant mutants, the atpE gene, which encodes a transmembrane and oligomeric C subunit of the ATP synthase and which was previously shown to be involved in resistance, was sequenced. For 15/53 mutants, five different point mutations resulting in five different amino acid substitutions were identified in the atpE gene. For 38/53 mutants, no atpE mutations were found and sequencing of the complete F0 ATP synthase operon (atpB, atpE, and atpF genes) and the F1 ATP synthase operon (atpH, atpA, atpG, atpD, and atpC genes) from three mutants revealed no mutations, indicating other, alternative resistance mechanisms. Competition assays showed no measurable reduction in the fitness of the mutants compared to that of the isogenic wild types.

Sub-acute toxicity evaluation of an aqueous extract of Labisia pumila, a Malaysian herb.

Labisia pumila (Myrsinaceae), is a popular herb among the women in Malaysia known locally as "Kacip Fatimah". Recently many nutraceutical products containing the powdered or extracted parts of the plant have become available for women's health care. However no evaluation of the effect of the repeated dosing of any herbal product of this plant had been undertaken prior to a 28-day sub-acute study presented in this report. The results showed that a dose of 50mg/kg of an aqueous extract of L. pumila corresponded to no-adverse-effect-level (NOAEL), whereas higher doses were associated with some toxicity concerns.

Potential of Azadirachta indica against Salmonella typhimurium-induced inflammation in BALB/c mice.

OBJECTIVE: To evaluate the potential of Azadirachta indica leaf extracts against Salmonella typhimurium-induced inflammation in BALB/c mice. DESIGN: Qualitative tests of A. indica leaf extracts were conducted for screening of various phytochemicals. The antiinflammatory potential of A. indica leaf extracts on S. typhimurium and its outer membrane proteins (OMPs)-induced inflammation was assessed by hyperalgesic (flicking) response of the mice inflamed paws. The monokines (IL-1alpha, IL-6 and TNF-alpha) activities in the culture supernatants of macrophages (infected with bacteria and interacted with OMPs) in the presence or absence of A. indica leaf extracts was assessed by ELISA. RESULTS: Aqueous and petroleum ether A. indica leaf extracts reduced the inflammation caused by S. typhimurium and its OMPs as assessed by paw flicking response. Petroleum ether A. indica leaf extract was found to be more effective than aqueous A. indica leaf extract. Significantly lower levels of monokines (IL-6 and TNF-alpha) were also observed in the presence of petroleum ether A. indica leaf extracts than aqueous A. indica leaf extract. These observations may be due to the presence of steroids and triterpenoids observed in petroleum ether extract. CONCLUSION: Petroleum ether A. indica leaf extract seems promising to combat S. typhimurium-induced inflammation.