Fibrinolytic and haemostatic activities of caffeic acid phenethyl ester and propolis from Malaysian stingless bee and romanian poplar in vitro

Caffeic acid phenethyl ester (CAPE) is a phenolic derivative from propolis and plants. The aims of this study include: (1) Development and validation of an in vitro whole blood (WB) clot lysis procedure for fibrinolytic activity study, (2) Investigation of fibrinolytic, antiplatelet and anticoagulant properties of CAPE, Malaysian Tetratrigona itama (T.itama) and poplar propolis and (3) Determination and quantification of CAPE compound in Malaysian T.itama propolis. The WB clot lysis procedure was developed using a standardized unresected retracted WB clot incubated in pooled platelet poor plasma (PPP) for varying incubation times and successfully validated using streptokinase (SK). The fibrinolyic activity was assessed by D-Dimer (DD), fibrin morphology by confocal microscopy and WB clot weight. DD was measured photometrically by immuno-turbidometric method. Fibrinolytic activity of CAPE, Malaysian T.itama and poplar propolis was assessed by the new WB clot lysis procedure at different concentrations and different times. Platelet activity study of CAPE was performed using different in vitro assays including: 1) Platelet aggregation measurement by platelet aggregometry with different types of agonists, adenosine diphosphate (ADP), arachidonic acid (AA) and ristocetin. 2) Platelet activation markers (PAC-1 and P-selectin) expression by flow cytometry. 3) P2Y12 receptor determination by Western blot (W.blot) technique. The platelet study of Malaysian T.itama and poplar propolis was done by platelet aggregometry. Thromboelastgraphy (TEG) parameters were recorded following WB incubation with CAPE, Malaysian T.itama and poplar propolis. Quantitation of CAPE in propolis was performed by Gas Chromatograph-Mass Spectrometer (GCMS) analysis using constructed calibration curves on peak area versus various concentrations of CAPE. The mean differences of DD (μg/ml) levels were significantly different (p<0.05) across samples incubated with different CAPE concentrations and both types of propolis compared with normal control (PPP). The median pre and post-incubation WB clot weights (gm) were significantly decreased for CAPE, Malaysian T.itama and poplar propolis. Fibrin removal was observed microscopically and indicated dose-dependent effects of CAPE compared with that of normal control at different time. The 50% effective dose (ED50) of CAPE (based on DD) was 1.99 mg/ml. The anti-platelet effect was observed by the techniques used in this study for CAPE and both propolis. The ED50 of CAPE, Malaysian T.itama and poplar propolis (based on platelet aggregation) was 7.31 μg/ml, 0.79 mg/ml and 0.86 mg/ml respectively. TEG results showed fibrinolytic parameter (LY30), was significantly different (p<0.17) from normal control samples incubated in different concentrations of CAPE, Malaysian T.itama and poplar propolis. The antiplatelet effect of CAPE may have contributed to the reduced maximum amplitude (MA) value of TEG assay. The quantity of CAPE measured in Malaysian T.itama and poplar propolis were: 0.6 (0.1) and 29.7 (1.2) mg/g respectively

In Vitro

This study aimed to evaluate in vitro whole blood (WB) clot lysis method for the assessment of fibrinolytic activity. Standardized unresected (uncut) retracted WB clot was incubated in pool platelet poor plasma (PPP) for varying incubation times and in streptokinase (SK) at different concentrations. The fibrinolytic activity was assessed by D-dimer (DD), confocal microscopy, and clot weight. DD was measured photometrically by immunoturbidimetric method. There was a significant difference in mean DD levels according to SK concentrations (P=0.007). The mean DD±SD according to the SK concentrations of 5, 30, 50, and 100 IU/mL was: 0.69±0.12, 0.78±0.14, 1.04±0.14 and 2.40±1.09 μg/mL. There were no significant changes of clot weight at different SK concentrations. Gradual loss and increased branching of fibrin in both PPP and SK were observed. Quantitation of DD and morphology of fibrin loss as observed by the imaging features are in keeping with fibrinolytic activity. Combination of DD levels and confocal microscopic features was successfully applied to evaluate the in vitro WB clot lysis method described here

The concepts of distinctiveness and similarity in the trade marks law of the UK

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Impact of Sensitization of Family Caregivers upon Treatment Compliance among Geriatric Patients Suffering from Elder Abuse and Neglect

Geriatric patients in various outpatient department (OPDs) have been found to agonize from elder abuse and neglect (EAN). Such suffering imposes depressive states within individuals, which in turn affects treatment compliance. The objective of this study was to evaluate the impact of sensitization (psychotherapeutic) of family caregivers (FCGs) upon two denture treatment parameters (maintenance and treatment satisfaction) among EAN patients and compare the differences in outcome with non-abused patients. A survey of completely edentulous subjects (n = 860, aged 41–80 years) provided a sampling frame of 332 EAN patients from which 150 patients (including FCGs) fulfilling the study criteria were distributed (simple random, convenient) into two groups (Group A—control, Group B—test). FCG sensitization for subjects in Group B was performed by a clinical psychologist in 2–4 short (30 min) sessions. Demographic characteristics (frequency) were measured using a self-reported questionnaire, denture maintenance was measured using a denture hygiene index (scores), and treatment satisfaction was analyzed on a 10-point visual analog scale. Relevant data were calculated for means and absolute/relative frequencies. Any difference between two groups was estimated using an unpaired t-test while the level of relationship was determined by Karl Pearson’s test at a p-value of < 0.05. The results showed highest frequency (38.6%) for neglect, with elder neglect (EN) being most common (38.14% alone and 14% in combination). EN was found more if the FCG was a son (52%), in the age group (21–30 years), and with low education and low income (75%). Patients whose FCGs were counselled (Group B) demonstrated low denture plaque scores (mean = 1.38 ± 0.618), while demonstrating comparatively higher scores in six different parameters of treatment satisfaction. Differences between the two groups for both parameters were also found to be statistically significant. Psychotherapeutic counselling in the form of FCG sensitization brings better results of denture maintenance and treatment satisfaction

Modified micromarsupialization technique as an alternative primary treatment for ranulas: A case series in a resource‐challenged economy

Abstract Objectives Ranula is one of the commonest salivary gland cysts that mostly occur due to mucus extravasation from the sublingual salivary gland. Treatment of this lesion is still somewhat shrouded in controversy and varies from conservative treatment to surgical excision of the causative gland. Materials and methods This was a case series in quasi‐experimental design that evaluated the outcome and complications of the modified micromarsupialization technique as a newly introduced treatment at our center for simple sublingual ranula over a 2‐year period. Results Twenty patients were treated by the modified micromarsupialization technique and followed up for 1 year. There was complete resolution following this technique in 17 patients (85%) with no evidence of recurrence or complications, whereas 2 patients (10%) showed partial resolution and the remaining 1 patient (5%) showed a failure and recurrence. The age of the patient, the size of the ranula, and the retention of sutures throughout the study period did not significantly affect the treatment outcome. Conclusions The modified micromarsupialization technique was a simple and effective treatment that should be used as a primary treatment option for simple ranulas and we recommend it to be the first‐choice treatment before surgical excision of the sublingual gland, especially in a resource‐challenged economy like ours

Fibrinolytic Activity and Dose-Dependent Effect of Incubating Human Blood Clots in Caffeic Acid Phenethyl Ester: In Vitro Assays

Background. Caffeic acid phenethyl ester (CAPE) has been reported to possess time-dependent fibrinolytic activity by in vitro assay. This study is aimed at investigating fibrinolytic dose-dependent activity of CAPE using in vitro assays. Methods. Standardized human whole blood (WB) clots were incubated in either blank controls or different concentrations of CAPE (3.75, 7.50, 15.00, 22.50, and 30.00 mM). After 3 hours, D-dimer (DD) levels and WB clot weights were measured for each concentration. Thromboelastography (TEG) parameters were recorded following CAPE incubation, and fibrin morphology was examined under a confocal microscope. Results. Overall, mean DD (μg/mL) levels were significantly different across samples incubated with different CAPE concentrations, and the median pre- and postincubation WB clot weights (grams) were significantly decreased for each CAPE concentration. Fibrin removal was observed microscopically and indicated dose-dependent effects. Based on the TEG test, the Ly30 fibrinolytic parameter was significantly different between samples incubated with two different CAPE concentrations (15.0 and 22.50 mM). The 50% effective dose (ED50) of CAPE (based on DD) was 1.99 mg/mL. Conclusions. This study suggests that CAPE possesses fibrinolytic activity following in vitro incubation and that it has dose-dependent activities. Therefore, further investigation into CAPE as a potential alternative thrombolytic agent should be conducted