Activity of Fluoroquinolones and Proton Pump Inhibitors against Resistant Oral Bacterial Biofilms, in silico and in vitro Analysis.

Oral bacterial infections are a great health concern worldwide especially in diabetic patients. Emergence of antimicrobial resistance with reference to biofilms in oral cavity is of great concern. We investigated antibiotics combination with proton pump inhibitors against oral clinical isolates. The strains were identified as Staphylococcus epidermidis and Staphylococcus aureus by the 16S rRNA gene sequencing. In molecular docking, ciprofloxacin, levofloxacin, and omeprazole best fit to active pockets of transcriptional regulators 4BXI and 3QP1. None of the proton pump inhibitors were active against S. epidermidis, whereas omeprazole showed significant inhibition (MIC 3.9 µg/ml). Fluoroquinolones were active against both S. epidermidis and S. aureus. In combination analysis, a marked decrease in minimum inhibitory concentration was noticed with omeprazole (MIC 0.12 µg/ml). In antiquorum sensing experiments, a significant inhibitory zone was shown for all fluoroquinolones (14-20 mm), whereas among proton pump inhibitors, only omeprazole (12 ± 0.12 mm) was active against Chromobacterium violaceum. In combination analysis, a moderate increase in antiquorum sensing activity was recorded for ciprofloxacin, ofloxacin, and proton pump inhibitors. Further, significant S. aureus biofilm eradication was recorded using of ciprofloxacin, levofloxacin, and omeprazole combination (78 ± 2.1%). The time-kill kinetic studies indicated a bactericidal effect by ciprofloxacin: levofloxacin: omeprazole combination over 24 hrs. It was concluded that fluoroquinolone combined with omeprazole could be an effective treatment option for eradicating oral bacterial biofilms.

Radon concentration in spring water as an indicator of seismic activity: a case study of the Muzaffarabad Fault in Pakistan.

Radon and its progenies found in water indicate the existence of seismically active faults in the region. However, exposure to high levels of radon can also result in radiation-related health risks. This study focuses on radon-based active tectonic studies along the Muzaffarabad Fault in the core of the Hazara-Kashmir Syntaxis (HKS), NW Himalayas, Pakistan. In this study, spring water samples were collected along roadside of Jhelum Valley and in close proximity to the Muzaffarabad Fault in Pakistan using Radon Thoron Monitor (RTM1688-2). The results of the study showed that the radon concentrations in the water samples ranged from 1.895 to 17.097 Bq/l. The study found that the highest radon concentration was observed in the samples collected closest to the fault, while the lowest concentration was observed in the samples collected further away. The statistical analysis between the radon concentration and the distance from the fault showed a strong inverse relationship (R2=0.73). The study also found that 68% of the sampling sites had radon concentrations that exceeded the maximum contamination level (MCL) set by the US Environmental Protection Agency (EPA). The higher radon concentrations in the springs water suggest the probability of earthquake, which in turn poses potential health risks for the local population. The findings suggest that the measurement of radon concentration in water can be used as a tool for identifying seismically active faults in the region.

Deep venous thrombosis post mechanical thrombectomy in acute ischaemic stroke.

A woman in her 20s presented with neck pain, slurred speech and left-sided weakness. She successfully underwent mechanical thrombectomy (MT) for total occlusion of the basilar artery and made full neurological recovery. Initial ultrasound Doppler of legs (USDL) did not show any deep vein thrombosis (DVT). However, 2 days later, a repeat USDL revealed an extensive proximal DVT in the right common femoral vein. CT scan confirmed a haematoma arising from the femoral access site causing compression of the adjacent common femoral vein. This may have initiated DVT formation or exacerbated extension of a previously undetected DVT. Echocardiogram revealed a patent foramen ovale. As the patient was asymptomatic, delayed or missed diagnosis of DVT would have put her at risk of life-threatening pulmonary embolism or further strokes. Clinicians should maintain a high index of suspicion of DVT as a direct procedural complication from MT so treatment can be initiated promptly.

In vitro evaluation of herbal based Lesh Nat B cream against Leishmania tropica.

Pentavalent antimonials continue to be the standard treatment for cutaneous leishmaniasis. But their use is retarded owing to highly-priced, prolonged hospitalization, noxious and poor solubility. Therefore, there is a dire need to characterize new potential compounds possessing anti-leishmanial activity. Topical therapies that are more successful are an essential alternative therapeutic option for the localized self-limiting form of this disease. We tested the herbal-based topical cream Lesh Nat B against Leishmania tropica KWH23 promastigotes and axenic amastigotes in vitro. The anti-leishmanial activity of Lesh Nat B cream was evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay against promastigotes and axenic amastigotes. The results of Lesh Nat B cream were concentration and incubation time-dependent. After 72 h of incubation, Lesh Nat B cream efficiently suppresses the promastigote form of the parasite, followed by 48 h and 24 h. At 72 h, the lowest and highest levels of activity were 37% and 90%. Amastigotes had a minimum activity of 34% and a maximum activity of 78.5%, respectively. This formulation was more cytotoxic against promastigote form than amastigotes form at 72 h incubation periods. All the experiments were carried out in triplicates. Half-maximal inhibitory concentration (IC50) values were determined to be (66 ug/ml) and (70 ug/ml) against promastigote and amastigote forms, respectively. Moreover, 1.63% hemolytic activity was observed in Lesh Nat B cream at (10 µg/ml) while 3% hemolytic activity was observed at (37 µg/ml). It can be concluded that Lesh Nat B cream demonstrated effective Leishmanicidal and less hemolytic activity and can be used as an alternative therapeutic option for the treatment of cutaneous leishmaniasis; however, more studies are expected to justify its effectiveness in treating cutaneous leishmaniasis in both humans and animals.

Novel Schiff bases of Vanillin: potent inhibitors of macrophage harbored Leishmania tropica.

Due to limited chemotherapeutic options for leishmaniasis, novel synthetic compounds are gaining attention for evaluation against leishmaniasis. This study aimed to synthesize the compound's Schiff bases of Vanillin to investigate and evaluate their anti-leishmanial potentials against intracellular protozoan parasites Leishmania tropica. In the current study, the phenomena of synergism by designing Schiff bases with Vanillin enhances their desired importance. A total of five compounds Schiff bases of Vanillin were synthesized using different aromatic amines and Vanillin. The structural analysis of all the compounds was done through FT-IR (Fourier Transformer-Infrared), thin layer chromatography, and spectroscopic techniques such as 13C-NMR, mass spectrometry, and 1H-NMR. The antimicrobial properties of all the compounds ZI-1, ZI-2, BS-1, KH-1, and FA-2 against promastigotes and amastigotes forms of L. tropica were analyzed at three different concentrations 25, 50, and 100 µg/ml. The in-vitro MTT assay was performed to calculate the percent inhibition, IC50 values, and their cytotoxicity. The highest percent inhibition values against promastigote form of L. tropica were BS-1 53.78% at 25 µg/ml, ZI-2 66.95% at 50 µg/ml, and again ZI-2 76.92% at 100 µg/ml. Similarly, the highest percent inhibition values against intracellular amastigote stage were BS-1 55.77% at 25 µg/ml, ZI-2 67.78% at 50 µg/ml and again ZI-2 84.93% 100 µg/ml. The highest potency was recorded for BS-1 in both stages, with IC50 values of 9.83 and 4.27 µg/ml against promastigotes and intracellular amastigotes, respectively. The percent hemolysis as toxicity; the lowest percent hemolysis was recorded for ZI-1 at three different concentrations of 25, 50, 100 µg/ml of 2.60, 3.50, and 6.31, respectively. These results suggested that all the compounds exhibited anti-leishmanial activity, with BS-1 as the most potent. Further studies are suggested to increase the activity of compounds with structural modifications by the addition of some other synergistic, novel, and analogue compounds.

Inhibitive efficacy of Nymphoides indica rhizome extract on α-glucosidase, and cross-link formation of advanced glycation end product.

OBJECTIVE: To investigate the inhibitive efficacy of Nymphoides Indica (L.) Kuntze rhizome extract on α-glucosidase and on cross-link formation of advanced glycation end products (AGEs). METHODS: The plant extracts were prepared by cold maceration and fractionated in solvents of diverse polarity. The in vitro α-glucosidase inhibition assay, fluorescence spectrometry and SDS-PAGE analysis was performed for antiglycation assays. RESULTS: During α-glucosidase inhibition assay significant inhibition by chloroform (0.43 mg/mL) and methanol fractions (0.66 mg/mL) was noticed. During the AGEs inhibition assay, both oxidative (BSA-MGO) and non-oxidative (BSA-glucose) modes were employed. The inhibition of AGEs by total extract was considered moderate (IC50 0.10 mg/mL) as a result of non-oxidative mode, whereas in case of oxidative mode (BASA-MGO) no activity was recorded. Among fractions the methanolic fraction presented significant results both in oxidative (IC50 0.01 mg/mL) and non-oxidative modes (IC50 0.3 mg/mL). Likewise the ethyl acetate fraction was more active in non-oxidative mode (IC50 0.04 mg/mL) compared to oxidative mode (IC50 0.32 mg/mL). During assay for inhibition of cross-link formation, the chloroform fraction significantly inhibited cross-link formation in a dose dependent mode. CONCLUSION: It was finally concluded that N. Indica rhizome extract possesses significant properties that inhibit α-glucosidase, and AGEs cross-link formation.

Studies on effects of umbelliferon derivatives against periodontal bacteria; antibiofilm, inhibition of quorum sensing and molecular docking analysis.

OBJECTIVE: Umbelliferon derivatives are exclusively found in plants of Ferula spp. that are commonly used in curing various health concerns related to oral cavity. Diabetic patient are especially effected with periodontitis and allied complications. METHOD: We investigated various compounds isolated from Ferula narthex exudate against clinical strains obtained from diabetic patients with periodontitis. Further antibiofilm, antiquorum sensing and molecular docking studies and ADMET analysis were performed. RESULTS: The docking target included 2Q0J, 2UV0, 3QP5 and 3QP1. HYDE affinity assessment was performed for the first 30 top ranking docked conformations within these active sites. The binding free energy ΔG, FlexX docking score and the most favorable poses for all the compounds were determined. During in vitro analysis, feselol presented high inhibition of Pseudomonas aeruginosa (MIC 0.01 mg/mL, MBC 0.02 mg/mL). Similarly, Feselol presented significant inhibition against clinical strain S. epidermidis (MIC 0.087 mg/mL, MBC 0.174 mg/mL) and S. aureus (MIC 0.087 mg/mL, MBC 0.087 mg/mL) preceded by 10'-R-acetyl-karatavacinol against S. epidermidis (MIC 0.56 mg/mL, MBC 0.56 mg/mL) and S. aureus (MIC 0.28 mg/mL, MBC 0.28 mg/mL). During antibiofilm inhibition assay, 10' R-acetyl-karatavacinol showed significant inhibition (54% at a final concentration 0.45 mg/mL), whereas slight antiquorum sensing activity was recorded. CONCLUSIONS: The umbelliferon derivatives have significant inhibition of clinical isolates and moderate antibiofilm potential.

Antibacterial and antibiofilm properties of traditional medicinal plant from Sheikh Buddin range.

During current project, antibacterial and antibiofilm properties of traditional medicinal plant Ziziphus nummularia leaf extract and various fractions was investigated. The plant leaves were dried and extracted using 90% methanol followed by sequential fractionation using liquid-liquid fractionation. The fractions of a diverse polarity including chloroform, n-hexane, methanol and ethyl acetate and aqueous extracts were obtained that was further analysed by using HPLC. The phytochemical screening indicated presence of saponins, triterpenes and flavonoids. During DPPH assay, the methanolic fraction presented highest activity (IC50 193.1µg/mL), followed by ethyl acetate (IC50 220µg/mL) and chloroform (IC50 263µg/mL) fractions respectively. During FRAP assay, FRAP value for Z. nummularia extract 20.43µM. Among fractions, ethyl acetate fraction presented highest FRAP value (370.2µM), followed by chloroform (204µM) and methanolic (249µM) fractions. The antimicrobial activity of chloroform fraction was significantly high against P. aureginosa (6mm), L. monocytogenes, S. aureus (5mm), K. pneumoniae, B. Subtillus and E. coli (4mm). The ethyl acetate part presented significant activity (MIC 4mg/mL) against S. aureus, B. Subtillus and L. monocytogenes. The total extract and fractions were further tested for MBC and the MBC for ethyl acetate fractions was 4mg/mL, whereas all other fractions exhibited MBC >10mg/mL. No activity was recorded against Aspergillus niger. During antibiofilm assay, n-hexane fraction presented highest inhibition (88%) followed by ethyl acetate (69%) chloroform (65%) fractions. It was concluded that Z. nummularia possess moderate antimicrobial and antibiofilm activities. Further a synergistic effect is suggested in formulation having Z. nummularia.

A Pilot Study Assessing the Impact of 3-D Printed Models of Aortic Aneurysms on Management Decisions in EVAR Planning.

INTRODUCTION: Endovascular repair of aortic aneurysms with difficult anatomy is challenging. There is no consensus for planning such procedures. METHODS: Six cases of aortic aneurysms with challenging anatomical features, such as short, angulated, and conical necks and tortuous iliacs were harvested. The computed tomography (CT) scans were anonymized. Lifesize 3-dimensional (3-D) printed models were created of the lumen. Endovascular operators were asked to review the CT angiography (CTA), make a management plan, and give an indication of their confidence. They were then presented with the equivalent model and asked to review their decision. Their attitudes to such models were briefly surveyed. RESULTS: A total of 28 endovascular operators reviewed 144 cases. After review of the physical model, the management plan changed in 29 (20.1%) of 144 cases. Initial plan after CTA review was endovascular 73.6%, open repair 22.9%, and second opinion 3.5%. After model review, this became endovascular 67.4%, open repair 19.4%, and second opinion 4.8%. Although the general trend was toward more open procedures, off-label techniques reduced from 19.4% to 15.2% following model review. When the management plan did not change, level of confidence did increase in 37 (43.5%) of 85 cases. The majority of operators stated that they would find models useful for planning in some procedures. For 1 case, the change in the percentage of participants being sure in the management plan was statistically significant (P = .031). CONCLUSION: The 3-D printed models may be potentially useful in planning cases with EVAR. It is a paradigm that warrants further investigation.

Functional lengths of the lower limb arterial tree measured by CT angiography centreline analysis: implications for catheter lengths in peripheral angioplasty.

PURPOSE: Lower limb angioplasty is a common procedure. However, arterial lengths have not been well studied and there is no evidence base for the optimum catheter lengths required for the various applications of femoral or distal below-the-knee angioplasty. The industry standard catheter measures 80 cm. METHOD: Fifty CT angiograms were post-processed using vessel tracking and centreline analysis tools and lengths were measured from the ipsilateral first segment of the femoral artery (FSFA) (common femoral artery) to the contralateral FSFA and on to the second segment of the femoral artery (superficial femoral artery) and popliteal arteries down to the posterior tibial (PT) artery at the ankle. This allowed clinically meaningful lengths for 'cross-over' and 'antegrade' angioplasty to be calculated. RESULTS: Mean cross-over length to the second segment of the femoral artery as it crossed the femoral cortex was 72.3 cm, and the mean cross-over length to the popliteal artery at the knee joint was 83.8 cm, and the length from the FSFA to the PT was 85.1 cm. CONCLUSION: Selection of a standard length catheter can result in a situation where the catheter is too short. Optimum catheter length for a particular task will reduce the need for catheter exchanges and use of multiple balloons and therefore reduce complications, procedure time, radiation dose and cost.