Antibacterial, antioxidant and hypoglycaemic effects of Thymus capitatus (L.) Hoffmanns. et Link leaves' fractions.

The aim of this study was to examine the bioactivity of the methanol fraction (MF) and n-hexane fraction (HF) of Thymus capitatus leaves in relation to their constituents analysed by gas chromatography and gas chromatography-mass spectrometry. The effects of T. capitatus on the growth of pathogenic bacteria associated with respiratory diseases (13 gram-positive and 4 gram-negative) were determined using a microdilution method. The MF was particularly effective on Streptococcus pneumoniae and Moraxella catarrhalis. The antioxidant activity was evaluated by 2,2-diphenyl-1-picrylhydrazyl, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid), ferric-reducing antioxidant power and β-carotene bleaching assays. A strong activity using β-carotene bleaching test was observed with the MF (IC50 of 0.7 μg/mL after 30 min of incubation). In the hypoglycaemic test, a selective α-amylase inhibitory activity was detected with the HF begging the most active (IC50 of 422.5 μg/mL). T. capitatus may represent a source of natural bioactive compounds

Use of the T-spot.TB test for the diagnosis of latent tuberculosis infection

Background:Tuberculosis (TB) represents a major health problem both in developing and both in industrialized countries.The identification of individuals latently infected with Mycobacterium tuberculosis (Mtb) play a key role for the efficacy of TB control. These individuals with a latent tuberculosis infection (LTBI), especially those with high risk of reactivation (e.g. HIV + / AIDS-infected individuals, patients undergoing immunosuppressive therapy and children younger than 5 years) could benefit from a preventive treatment with isoniazid reducing the risk of progression from LTBI to active TB. Until recently, detection of LTBI has relied on the tuberculin skin test (TST), but despite the widespread use in clinical practice,TST does not reliably diagnose LTBI because several drawbacks, e.g. lacking in specificity, particularly in who were exposed to non-tuberculous mycobacteria (NTM) or were vaccinated with Bacille Calmette-Guerin (BCG) In addition, in young subjects,TST sensitivity is hampered by impaired T cell function leading frequently to false negative results.These several drawbacks limit the use of TST for the diagnose an LTBI in patients who may benefit from preventive chemotherapy. On the other hand, an accurate diagnosis of LTBI avoid the over-treatment of those patients with a positive TST results but not latently infected with Mtb. Recently, new tests based on the detection of interferon-gamma (IFN-γ) after stimulation with Mtb-specific antigens: Early secretory Antigenic Target-6 (ESAT-6) and Culture Filtrate Protein-10 (CFP-10) have been proposed for the diagnosis of active TB and LTBI. Methods: During the period from January 2009 to June 2009, in our laboratory 70 patients were tested with T-SPOT.TB (Oxford Immunotech, Abingdon, United Kingdom).We enrolled transplant patients and subjects ongoing transplant, patients immigrants from high prevalence TB countries, patients screened for immunosuppressive treatment, HIV / AIDS – infected individuals.We also tested 3 patients with clinical / radiological suspicion of active TB and 3 patients with positive tuberculin skin test and with a positive direct examination for mycobacteria in the urinary sediment. Results: In 2 patients with symptoms suggestive of TB in place,T-SPOT.TB showed a higher response of (IFN-g), more than 100 spots.Among individuals ongoing renal transplant, 6 patients tested T-SPOT.TB positive and 4 subjects were T.SPOT.TB -negative. Two patients with an autoimmune disease showed an high response to Mtb-specific antigens with T-SPOT.TB test tested before to start any treatment.T-SPOT.TB test tested strongly negative in 4 paediatric patients and in one HIV-infected individuals, regardless a positive response to a internal positive response (phytohaemagglutinin (PHA), suggesting a normal immune response. Conclusions:This preliminary data suggest that the T.SPOT.TB showed high sensitivity and specificity, producing a strongly negative response to Mtb-specific antigens in subjects who had a history of previous BCG-vaccination. In addition, T-SPOT.TB test provides, unlike the TST, indication about the potential immunosuppression of tested patient with an internal positive control that can highlight the production of IFN- γ by lymphocytes resulting in the application of this test in immunocompromised patients, e.g. children and transplantated patients and others

Antioxidant Properties of Berberis aetnensis C. Presl (Berberidaceae) Roots Extract and Protective Effects on Astroglial Cell Cultures

Berberis aetnensis C. Presl (Berberidaceae) is a bushy-spiny shrub common on Mount Etna (Sicily). We demonstrated that the alkaloid extract of roots of B. aetnensis C. Presl contains prevalently berberine and berbamine, possesses antimicrobial properties, and was able to counteract the upregulation evoked by glutamate of tissue transglutaminase in primary rat astroglial cell cultures. Until now, there are no reports regarding antioxidant properties of B. aetnensis C. Presl collected in Sicily. Air-dried, powdered roots of B. aetnensis C. Presl were extracted, identified, and quantified by HPLC. We assessed in cellular free system its effect on superoxide anion, radicals scavenging activity of antioxidants against free radicals like the 1,1-diphenyl-2-picrylhydrazyl radical, and the inhibition of xanthine oxidase activity. In primary rat astroglial cell cultures, exposed to glutamate, we evaluated the effect of the extract on glutathione levels and on intracellular production of reactive oxygen species generated by glutamate. The alkaloid extract of B. aetnensis C. Presl inhibited superoxide anion, restored to control values, the decrease of GSH levels, and the production of reactive oxygen species. Potent antioxidant activities of the alkaloid extract of roots of B. aetnensis C. Presl may be one of the mechanisms by which the extract is effective against health disorders associated to oxidative stress

Effect of berberine and Berberis aetnensis C. Presl. alkaloid extract on tissue transglutaminase in primary astroglial cell cultures

International audienceBerberis aetnensis C. Presl. is a bushy-spiny shrub common on Mount Etna (Sicily, Italy), containing various alkaloids with several pharmacological properties. Herein, we assessed the effect of berberine and of the alkaloid extract of B. aetnensis roots on the glutamate-evoked tissue transglutaminase (TG2) up-regulation in rat astrocyte primary cultures, used as an in vitro model of excitotoxicity. Our findings show that the alkaloid extract of B. aetnensis roots consists mainly of berberine. Furthermore, berberine and the alkaloid extract of B. aetnensis roots were able to restore the oxidative status modified by glutamate and the levels of TG2 to control values. We found that berberine or the alkaloid extract of B. aetnensis roots are able to ameliorate excessive production of glutamate, protein misfolding and aggregation, mitochondrial fragmentation, and neurodegeneration. Thus, we suggest that berberine and the alkaloid extract of B. aetnensis roots, may represent a natural therapeutic strategy in the neuropathological conditions associated to excitotoxicity

Syphilis serology: Seroprevalence in a selected population and considerations on the Euroline WB test

Introduction: The clinical diagnosis of syphilis is always supported by appropriate laboratory tests and the test results are interpreted with reference to the patient’s history. In the diagnosis of syphilis, the use of tests based on antibody search that recognize both treponemal and reaginic antigens increases the diagnostic chances. Our study discusses the various serological and alternative tests currently available along with their limitations, and relates their results to the likely corresponding clinical stage of the disease. Methods: in our laboratory were analyzed 264 sera and 4 liquor (123 Females, 145 Males). 187 patients are subject at low risk for luetic infection, including pregnant woman, patient with organ transplant, outpatients or hospitalized undergoing routine serological, and 81 from patients with confirmed syphilis including 4 pregnant women in antibiotic treatment, patients with suspected disease, HIV positive and patients with autoimmune diseases with Cardiolipin positive. All sera were tested with ELISA Anti-Treponema pallidum Screen (IgG / IgM) and in parallel with agglutination tests VDRL and TPHA. On all positive sera was tested Euroline-WB EUROIMMUN and reading done with the program EuroLineScan. Results: by ELISA Anti-Treponema pallidum Screen IgG / IgM 162 sera were negative and 106 sera positive (39.5%), distributed as follows: 45 (42%) with a value greater than 200 RU / ml, 43 (41% ) with a value> 22 RU / ml and 18 (17%) with a borderline value between> 16 to <22 RU / ml. The execution of the Blot IgG showed: 18 negative sera, 6 with borderline value with one only band of specific antigens (p15, p45, p47 or p17), while 82, including 4 liquor (neurolue), were certainly positive showing more than one band antibody to the treponemal antigens. Only one patient had in place at the time of screening, an initial infection; in fact, there was a single clear positivity in the IgM protein bands, while 7 sera was uncertain values. It is reported 11 positivity for IgM Cardiolipin, while Cardiolipin IgG was detected with a high positivity in 34 sera. The presence of borderline values and / or positivity for a single protein band can be attributed to a unspecific reaction caused by autoimmune diseases or related cross-reactions with other Spirochete or to other Borrelia. Conclusions: The immunoblot test gave useful information at epidemiological and clinical level. The deepening with a confirmation test with proteic antigens and cardiolipin identifies false reactivity, but also indicates the specific reactivity to past infection and a better characteriation in the different stages of disease. In our study in the latent forms there are relevant discrepancies among the various tests. Compare to traditional methods, anti-cardiolipin antibodies positivity in our confirmatory test has the advantage of providing non subjective interpretation, being based upon the EuroLineScan program

Use of the T-spot.TB test for the diagnosis of latent tuberculosis infection

Background:Tuberculosis (TB) represents a major health problem both in developing and both in industrialized countries.The identification of individuals latently infected with Mycobacterium tuberculosis (Mtb) play a key role for the efficacy of TB control. These individuals with a latent tuberculosis infection (LTBI), especially those with high risk of reactivation (e.g. HIV + / AIDS-infected individuals, patients undergoing immunosuppressive therapy and children younger than 5 years) could benefit from a preventive treatment with isoniazid reducing the risk of progression from LTBI to active TB. Until recently, detection of LTBI has relied on the tuberculin skin test (TST), but despite the widespread use in clinical practice,TST does not reliably diagnose LTBI because several drawbacks, e.g. lacking in specificity, particularly in who were exposed to non-tuberculous mycobacteria (NTM) or were vaccinated with Bacille Calmette-Guerin (BCG) In addition, in young subjects,TST sensitivity is hampered by impaired T cell function leading frequently to false negative results.These several drawbacks limit the use of TST for the diagnose an LTBI in patients who may benefit from preventive chemotherapy. On the other hand, an accurate diagnosis of LTBI avoid the over-treatment of those patients with a positive TST results but not latently infected with Mtb. Recently, new tests based on the detection of interferon-gamma (IFN-γ) after stimulation with Mtb-specific antigens: Early secretory Antigenic Target-6 (ESAT-6) and Culture Filtrate Protein-10 (CFP-10) have been proposed for the diagnosis of active TB and LTBI. Methods: During the period from January 2009 to June 2009, in our laboratory 70 patients were tested with T-SPOT.TB (Oxford Immunotech, Abingdon, United Kingdom).We enrolled transplant patients and subjects ongoing transplant, patients immigrants from high prevalence TB countries, patients screened for immunosuppressive treatment, HIV / AIDS – infected individuals.We also tested 3 patients with clinical / radiological suspicion of active TB and 3 patients with positive tuberculin skin test and with a positive direct examination for mycobacteria in the urinary sediment. Results: In 2 patients with symptoms suggestive of TB in place,T-SPOT.TB showed a higher response of (IFN-g), more than 100 spots.Among individuals ongoing renal transplant, 6 patients tested T-SPOT.TB positive and 4 subjects were T.SPOT.TB -negative. Two patients with an autoimmune disease showed an high response to Mtb-specific antigens with T-SPOT.TB test tested before to start any treatment.T-SPOT.TB test tested strongly negative in 4 paediatric patients and in one HIV-infected individuals, regardless a positive response to a internal positive response (phytohaemagglutinin (PHA), suggesting a normal immune response. Conclusions:This preliminary data suggest that the T.SPOT.TB showed high sensitivity and specificity, producing a strongly negative response to Mtb-specific antigens in subjects who had a history of previous BCG-vaccination. In addition, T-SPOT.TB test provides, unlike the TST, indication about the potential immunosuppression of tested patient with an internal positive control that can highlight the production of IFN- γ by lymphocytes resulting in the application of this test in immunocompromised patients, e.g. children and transplantated patients and others

Antioxidant Activity of Extracts of Momordica Foetida Schumach. et Thonn.

molecule

Effect of aqueous extract of <i>Tithonia diversifolia</i> (Hemsl.) A. Gray on LOOH production in plasma; results are expressed as percentage of inhibition of LOOH formation with respect to the same sample incubated in absence of the extract.

<p>Each value represents the mean ± SD of 5 experimental determinations.</p

Effect of aqueous extract of leaves of <i>Tithonia diversifolia</i> (Hemsl.) A. Gray on adipogenesis of hMSCs as measured by Oil-Red O staining.

<p>Each value represents the mean ± S.D. of 5 experimental determinations. * = p< 0.001 <i>vs</i> control.</p

Representative Western bloting of HO-1 and pAMPK: effect of aqueous extract of <i>Tithonia diversifolia</i> (Hemsl.) A. Gray on HO-1 and pAMPK expression in cultured hMSCs.

<p>Results, expressed as arbitrary units (A.U.), represent the mean ± S.D. of 5 experimental determinations. * = p< 0.005 with respect to tcontrol (C = hMSC control, T = hMSCs treated with 175 μmg/ml of aqueous extract of <i>Tithonia diversifolia</i> (Hemsl.) A. Gray).</p