The Effect of Pluchea indica (L.) Less. Tea on Adipogenesis in 3T3-L1 Adipocytes and Lipase Activity

Obesity and hyperlipidemia are a major problem in the world. Pluchea indica (L.) Less. tea (PIT) is a beverage that has various indications. This study focused on the effect of the PIT on inhibiting adipogenesis of 3T3-L1 cells and pancreatic lipase enzyme activity. The viability of 3T3-L1 cells was not significantly decreased after exposure to 200 to 1000 μg mL−1 PIT compared to controls (p < 0.05). The PIT at 750 to 1000 μg mL−1 exhibited a significantly reduced lipid accumulation compared to the control (p < 0.05). The inhibitory effects of the PIT at 250 to 1000 μg mL−1 on lipase activity were significantly increased compared to control (p < 0.05). The FTIR results showed that the integrated areas of lipids, proteins, nucleic acids, glycogen, and carbohydrates of the PIT-treated 3T3-L1 adipocytes were significantly lower than the untreated 3T3-L1 adipocytes (p < 0.05). These findings may indicate that the PIT is not only capable of inhibiting lipids and carbohydrate accumulation in adipocytes but also has a potential to inhibit pancreatic lipase activity. So, the PIT may be further developed to the novel lipid-lowering herbal supplement for the management of overweight or obesity

Mode of Action and Synergy of Ceftazidime and Baicalein against Streptococcus pyogenes

Purpose: To investigate the antibacterial activity of baicalein used alone, or in combination with ceftazidime, against Streptococcus pyogenes.Methods: Minimum inhibitory concentration (MIC), checkerboard assay parameters, and viability curves were determined for S. pyogenes DMST 30653, 30654, and 30655. Cytoplasmic membrane (CM) permeability technique, enzyme assays, transmission electron microscopy and Fourier transforminfrared microspectroscopy were used to investigate the changes in the bacterial biomolecules.Results: The MIC of ceftazidime and baicalein against all the S. pyogenes strains were 0.50 and &gt; 256.0 μg/ml, respectively. A synergistic effect against these strains was exhibited by the ceftazidime/baicalein combination (fractional inhibitory concentration index, &lt; 0.37). The results for the viable counts indicate that this synergistic activity was present. Baicalein exerted inhibitory activity against β-lactamase. Compared with the controls, combining baicalein with ceftazidime caused peptidoglycan and morphological damage, significantly increased CM permeability and protein concentrations, and decreased cellular fatty acid and nucleic acid concentrations.Conclusion: Baicalein is a potential synergistic adjunct to ceftazidime for the treatment of S. pyogenes infections.Keywords: Streptococcus pyogenes, Cytoplasmic membrane permeability, Baicalein, Ceftazidime, Synergistic activity, Fourier Transform-infrared microspectroscopy, Transmission electron microscop

Pluchea indica (L.) Less. Tea Ameliorates Hyperglycemia, Dyslipidemia, and Obesity in High Fat Diet-Fed Mice.

Pluchea indica (L.) Less. (P. indica) tea has been used for a health-promoting drink, especially in Southeast Asia. The effect of P. indica tea (PIT) on amelioration of hyperglycemia; dyslipidemia that was total cholesterol (TC), LDL-cholesterol (LDL-C), HDL-cholesterol (HDL-C), and triglyceride (TG); and obesity in high fat diet-induced (HFD) mice was investigated. Oral glucose tolerance test (OGTT) displayed that PIT at 400 and 600 mg/kg orally ameliorated hyperglycemia with a dose-dependent manner compared to the untreated group. Moreover, PIT at these dosages exhibited significantly lower TC, LDL-C, TG, and perigonadal fat weight in HFD treated mice compared to HFD mice (P 0.05). The PIT chemical analysis results demonstrated that PIT contained total phenolic content (TPC), total flavonoid content (TFC), 4-O-caffeoylquinic acid (4-CQ), 5-O-caffeoylquinic acid (5-CQ), 3,4-O-dicaffeoylquinic acid (3,4-CQ), 3,5-O-dicaffeoylquinic acid (3,5-CQ), 4,5-O-dicaffeoylquinic acid (4,5-CQ), beta-caryophyllene, and gamma-gurjunene that may play an important role in inhibiting hyperlipidemia and hyperglycemia. Also, histological analysis expressed that the mean area and amount of perigonadal fat adipocytes of PIT treated groups were significantly lower and higher than the HFD group (P 0.05). These results suggest that PIT does not become toxic to the kidney, liver, and blood. In conclusion, PIT has the potential to develop into healthy food supplement or medicine for the prevention and treatment of hyperglycemic, hyperlipidemic, and obese patients

Nitro-Carba test, a novel and simple chromogenic phenotypic method for rapid screening of carbapeneamase-producing Enterobacteriaceae.

OBJECTIVES: The present study developed Nitro-Carba Test (NCT), a rapid and simple chromogenic method for screening of Carbapenemase-producing Enterobacteriaceae (CPE). METHODS: The NCT was validated with a total of 31 carbapenemase-producing isolates (9 KPCs, 11 MBLs and 11 OXA-48s) and with 56 non-carbapenemase-producing strains. The assay relies on the hydrolysis of nitrocefin in the presence of carbapenems. The carbapenemases were extracted with lysis buffer prior to addition to wells with and without imipenem (IPM), meropenem (MEM) and ertapenem (ETP). Following the addition of nitrocefin, a change in colour from yellow to red, indicating carbapenemase production, was observed within 20min. The susceptibility profiles of each bacterial strain were also investigated. RESULTS: A NCT detected all 31 CPEs within a timeframe of only 10seconds to 12min. All carbapenemase producers hydrolyzed nitrocefin in all wells. No colour change in the wells with carbapenems was observed in non-carbapenemase producers. The sensitivity for all three carbapenems was 100%, while specificity of IPM, MEM and ETP were 64.29%, 91.07% and 100%, respectively. IPM, MEM and ETP against all carbapenemase-producing strains had MIC values ranging from 0.5 to ≥256μg/mL, 0.25 to ≥256μg/mL and 1 to ≥256μg/mL, respectively. OXA-48-producing isolates showed lower MIC values compared with producers of MBL and KPC. CONCLUSION: This assay is a promising method detecting CPE rapidly. The NCT is a simple and reliable method, capable of detecting CPE in even carbapenem-susceptible strains

The Free Radical Scavenging and Anti-Isolated Human LDL Oxidation Activities of Pluchea indica (L.) Less. Tea Compared to Green Tea (Camellia sinensis).

Tea is one of the most popular beverages in the world. Camellia sinensis tea (CST) or green tea is widely regarded as a potent antioxidant. In Thailand, Pluchea indica (L.) Less. tea (PIT) has been commercially available as a health-promoting drink. This study focused on free radical scavenging activities of PIT, and its ability to protect isolated human low-density lipoproteins (LDL) from oxidation by chemical agents. A preliminary study to investigate the antioxidant nature of PIT was undertaken. These included common antioxidant assays involving 2,2-Diphenyl-1-picrylhydrazyl (DPPH), 2,2-azinobis-(3-ethylbenzothiazoline)-6-sulfonic acid (ABTS), hypochlorous acid (HOCl), and its potential to scavenge peroxynitrite. In separated experiments, isolated human LDL was challenged with either 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH), copper (Cu2+), or 3-Morpholinosydnonimine hydrochloride (SIN-1) to induce LDL oxidation. PIT exhibited antioxidant activity in all test systems and performed significantly better than CST in both DPPH (P < 0.05; IC50PIT = 245.85 ± 15.83 and CST = 315.41 ± 24.18 μg/ml) and peroxynitrite scavenging assays. PIT at 75 μg/ml almost fully prevented the peroxynitrite over a 5 h period. Moreover, it displayed similar properties to CST during the antioxidation of isolated human LDL using AAPH, Cu2+, SIN-1, and hypochlorous acid scavenging assays. However, it revealed a significantly lower ABTS scavenging activity than CST (P < 0.05; IC50PIT = 30.47 ± 2.20 and CST = 21.59 ± 0.67 μg/ml). The main constituents of the PIT were identified using LC-MS/MS. It contained 4-O-caffeoylquinic acid (4-CQ), 5-O-caffeoylquinic acid (5-CQ), 3,4-O-dicaffeoylquinic acid (3,4-CQ), 3,5-O-dicaffeoylquinic acid (3,5-CQ), and 4,5-O-dicaffeoylquinic acid (4,5-CQ). In conclusion, caffeoyl derivatives in PIT could play an important role in potent antioxidant properties. So, it may be further developed to be antioxidant beverages for preventing atherosclerosis and cardiovascular diseases associated with oxidative stress

The performance of a resazurin chromogenic agar plate with a combined disc method for rapid screening of extended-spectrum-β-lactamases, AmpC β-lactamases and co-β-lactamases in Enterobacteriaceae.

A resazurin chromogenic agar (RCA) along with combined disc method has been developed as a promising method for rapid screening of extended-spectrum-β-lactamase (ESBL), AmpC β-lactamase, and co-production of ESBL and AmpC. Cefpodoxime (CPD) discs supplemented with and without clavulanic acid (CA), cloxacillin (CX), or CA+CX were evaluated against 86-molecularly confirmed β-lactamase-producing Enterobacteriaceae, including 15 ESBLs, 32 AmpCs, 9 co-producers of ESBL and AmpC, and 30 carbapenemase producers. The CA and CX synergy test successfully detected all ESBL producers (100% sensitivity and 98.6% specificity) and all AmpC producers (100% sensitivity and 96.36% specificity). This assay also exhibited a good performance in the screening for the co-existence of ESBL and AmpC (88.89% sensitivity and 100% specificity). The RCA assay is a simple and inexpensive method that allows observation of results within 7 h. It can be applicable in any microbiological laboratory, especially in the endemic areas of ESBL, AmpC, or co-β-lactamase-producing Enterobacteriaceae

Intracellular ROS Scavenging and Anti-Inflammatory Activities of Oroxylum indicum Kurz (L.) Extract in LPS plus IFN-gamma-Activated RAW264.7 Macrophages

Oroxylum indicum (L.) Kurz has been used as plant-based food and herbal medicine in many Asian countries. The aim of the present study was to examine the antioxidant and anti-inflammatory activities of O. indicum extract (O. indicum) in RAW264.7 cells activated by LPS plus IFN-γ. The phytochemical compounds in O. indicum were identified by GC-MS and LC-MS/MS. Five flavonoids (luteolin, apigenin, baicalein, oroxylin A, and quercetin) and 27 volatile compounds were found in O. indicum. O. indicum presented antioxidant activities, including reducing ability by FRAP assay and free radical scavenging activity by DPPH assay. Moreover, O. indicum also suppressed LPS plus IFN-γ-activated reactive oxygen species generation in RAW264.7 macrophages. It possessed the potent anti-inflammatory action through suppressing nitric oxide (NO) and IL-6 secretion, possibly due to its ability to scavenge intracellular ROS. The synchrotron radiation-based Fourier transform infrared (SR-FTIR) spectroscopy results showed the alteration of signal intensity and integrated areas relating to lipid and protein of the activated RAW264.7 macrophages compared to unactivated cells. This is the first report of an application of the SR-FTIR technique to evaluate biomolecular changes in activated RAW264.7 cells. Our results indicate that O. indicum may be used as a potential source of nutraceutical for the development of health food supplement or a novel anti-inflammatory herbal medicine

A nitrocefin disc supplemented with ertapenem for rapid screening of carbapenemase-producing Enterobacteriaceae

Reliable, simple and rapid methods for laboratory detection of carbapenemases are important for an appropriate antibiotic administration. A nitrocefin disc containing ertapenem for rapid screening of carbapenemase production among Enterobacteriaceae is developed in the present study. A total of 87 molecularly-confirmed Enterobacteriaceae including 31 carbapenemase producers and 56 non-carbapenemase producers, were tested with nitrocefin discs supplemented with and without ertapenem (20 μg/disc). Nitrocefin discs with ertapenem successfully discriminated all 31 carbapenemase and all non-carbapenemase producers within 30 minutes. The sensitivity and specificity of the method were 100%. The minimum inhibitory concentrations (MICs) of ertapenem against all carbapenemase-producing isolates ranged from 1 to ≥ 256 μg/mL. This simple test could help to minimize the treatment failure and control the dissemination of infections caused by carbapenemase-associated resistant bacteria. It is a promising approach that could be performed routinely in any laboratory

Boesenbergia rotunda (L.) Mansf. extract potentiate the antibacterial activity of some β-lactams against β-lactam-resistant Staphylococci.

OBJECTIVE: The purpose of this study was to investigate the effect of Boesenbergia rotunda (L.) Mansf. Extract (BRE) and peptidoglycan inhibitor antibiotics, either used alone or in combination against β-lactam resistant Staphylococci. METHODS: Antibacterial and synergistic activities of BRE alone and in combination with ampicillin (AMP), cloxacillin (CLX), cefazolin (CFZ) or vancomycin (VAN) were evaluated against two β-lactam-resistant Staphylococcus aureus (BRSA) and one β-lactam-resistant Staphylococcus epidermidis (BRSE). The activities were confirmed by killing curve assays. The preliminary antimicrobial action was elucidated by transmission electron microscopy (TEM) and cytoplasmic membrane (CM) permeability. RESULTS: All tested Staphylococci were inhibited by BRE at a MIC of 16μg/mL. Two BRSA strains showed high resistance to CLX, AMP, and CFZ, while BRSE was resistant to CLX and AMP. All tested isolates remained susceptible to VAN. The chequerboard assay demonstrated the fractional inhibitory concentration index (FICI) at 0.502 of the BRE and CLX combination against the BRSA strains. Killing curve determinations confirmed the antibacterial and synergistic activities. The TEM study revealed collapse of the CM in BRE-treated cells and damage of both CM and peptidoglycan in BRE plus CLX-treated cells. The CM permeability assay showed that either BRE or nisin alone, and BRE plus CLX significantly induced leakage of OD260-absorbing materials. CONCLUSIONS: The BRE potentiated β-lactams, particularly CLX against β-lactam-resistant Staphylococci by damaging the CM and peptidoglycan layer, leading to leakage of intracellular materials. The combination of BRE and β-lactams provides a potential way forward in developing a novel antistaphylococcal agent

Synergism and the mechanism of action of the combination of alpha-mangostin isolated from Garcinia mangostana L. and oxacillin against an oxacillin-resistant Staphylococcus saprophyticus

Globally, staphylococci have developed resistance to many antibiotics. New approaches to chemotherapy are needed and one such approach could be to use plant derived actives with conventional antibiotics in a synergestic way. The purpose of this study was to isolate α-mangostin from the mangosteen (Garcinia mangostana L.; GML) and investigate antibacterial activity and mechanisms of action when used singly and when combined with oxacillin against oxacillin-resistant Staphylococcus saprophyticus (ORSS) strains. The isolated α-mangostin was confirmed by HPLC chromatogram and NMR spectroscopy. The minimum inhibitory concentration (MIC), checkerboard and killing curve were determined. The modes of action of these compounds were also investigated by enzyme assay, transmission electron microscopy (TEM), confocal microscopic images, and cytoplasmic membrane (CM) permeabilization studies. Results The MICs of isolated α-mangostin and oxacillin against these strains were 8 and 128 μg/ml, respectively. Checkerboard assays showed the synergistic activity of isolated α-mangostin (2 μg/ml) plus oxacillin (16 μg/ml) at a fractional inhibitory concentration index (FICI) of 0.37. The kill curve assay confirmed that the viability of oxacillin-resistant Staphylococcus saprophyticus DMST 27055 (ORSS-27055) was dramatically reduced after exposure to isolated α-mangostin (2 μg/ml) plus oxacillin (16 μg/ml). Enzyme assays demonstrated that isolated α-mangostin had an inhibitory activity against β-lactamase in a dose-dependent manner. TEM results clearly showed that these ORSS-27055 cells treated with this combination caused peptidoglycan and cytoplasmic membrane damage, irregular cell shapes and average cell areas were significantly larger than the control. Clearly, confocal microscopic images confirmed that this combination caused considerable peptidoglycan damage and DNA leakage. In addition, the CM permeability of ORSS-27055 was also increased by this combination of actives. Conclusions These findings provide evidence that isolated α-mangostin alone has not only some activity but also shows the synergistic activity with oxacillin against ORSS-27055. The chromone and isoprenyl structures could play a significant role in its action. This synergistic activity may involve three mechanisms of action. Firstly, potential effects of cytoplasmic membrane disruption and increases permeability. Secondly, inhibit β-lactamase activity. Finally, also damage to the peptidoglycan structure. We proposes the potential to develop a novel adjunct phytopharmaceutical to oxacillin for the treatment of ORSS. Future studies require clinical trials to establish if the synergy reported can be translated to animals and humans