Shady Helmi Awwad

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Shady Helmi Awwad

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Research

Quantification of Caffeine and Chlorogenic Acid in Green and Roasted Coffee Samples Using HPLC-DAD and Evaluation of the Effect of Degree of Roasting on Their Levels

Authors : Shady Awwad, Reem Issa, Lilian Alnsour, Dima Albals, Idrees Al-Momani

Abstract : Chlorogenic acid and caffeine are among the important components in coffee beans, determining the taste and aroma. In addition, phenols and antioxidants content possess vital health values. The main aim of this study is to determine the levels of caffeine and chlorogenic acid in several coffee samples of different origins and degrees of roasting. The coffee samples were extracted using hot water. The levels of caffeine and chlorogenic acid were quantified using high-performance liquid chromatography (HPLC) equipped with a diode array detector, a reverse phase system, and an ODS column (C18). Total phenol and antioxidant contents were previously determined for the same samples. The results showed that the highest content of caffeine was found in the medium roasted coffee (203.63 mg/L), and the highest content of chlorogenic acid content was found in the green coffee (543.23 mg/L). The results demonstrated a negative correlation between the chlorogenic acid levels with the degree of roasting, while it showed a positive correlation between the caffeine levels with the degree of roasting till a certain point where the levels dropped in the dark roasted coffee. The origin of coffee samples did not show any effect on any of the measured variables. Antioxidant effects of coffee samples were largely determined by chlorogenic acid content.

Keywords : Caffeine; chlorogenic acid; coffee; extraction; high-performance liquid chromatography (HPLC); quantification

Quantification of Total Phenols and Antioxidants in Coffee Samples of Different Origins and Evaluation of the Effect of Degree of Roasting on Their Levels

Authors : Lilian Alnsour, Reem Issa, Shady Awwad, Dima Albals, Idrees Al-Momani

Abstract : Phenolic and antioxidant compounds have received considerable attention due to their beneficial effects on human health. The aim of this study is to determine the content of total phenols and antioxidants in fifty-two coffee samples of different origins, purchased from the Jordanian local market, and investigate the effect of the degree of roasting on the levels of these compounds. The coffee samples were extracted using the hot water extraction method, while Folin–Ciocalteu (FC) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay methods were used to analyze these compounds. The results showed that the highest content of total phenol (16.55 mg/g equivalent to GAE) was found in the medium roasted coffee, and the highest content of antioxidants (1.07 mg/g equivalent to TEAC) content was found in the green coffee. Only light and medium roasted coffee showed a significant correlation (p < 0.05, R2 > 0.95) between the average of total phenolic and antioxidant content. A negative correlation between the antioxidant content and the degree of roasting (p < 0.05, R2 > 0.95) were shown, while it did not correlate with phenolic contents. Previously, a positive correlation between antioxidant and chlorogenic acids content was observed, with no correlation between the origin of coffee samples nor heavy metal content, which was previously determined for the same coffee samples. These findings suggest that the antioxidant content for coffee extracts is largely determined by its chlorogenic acid content, rather than the coffee origin or total phenolic and heavy metals content.

Keywords : coffee, phenols, antioxidant, TEAC, roasting, GAE

Circulatory Leptin Levels as a Key Link in the Potential Association between Insulin Resistance and Vitamin D Deficiency: A Review Article

Authors : Mahmoud Abu-Samak, Beisan A Mohammad, Ibrahim Mosleh, Omar Abdel-Majeed, Abeer Braham, Shady H Awwad, Ahamed Abu-Zaiton, Abdel Rahman M. Tawaha

Abstract : Vitamin D deficiency (VDD) and diabetes mellitus (DM) are worldwide problems. Many recent studies have shown a high prevalence of T2DM and VDD in Mediterranean countries including Jordan. Remarkably, VDD is more prevalent in obese people with inverse association with 25-hydroxy vitamin D (25OHD). Nevertheless, randomized clinical trials (RCTs) point did not confirm that yet in particular the association with an obesity marker (leptin hormone). Leptin is involved in the insulin resistance (IR) pathogenesis and development of T2DM. Some RCTs showed that the treatment of VDD by 1,25OHD2D3 (VD3) may improve the control of diabetes and insulin sensitivity or decrease the risk of disease. Conversely, leptin levels were positively associated with 25OHD levels. Overall, taking into account the U-shaped curve, it seems that the association between VD3 supplementation is a dose dependent. In this context, it can be concluded that when VD intake is below, the serum leptin level will be low and vice versa VD supplementation may cause raising serum leptin as shown in this review. Therefore, we suggest further clinical trials wither to confirm or negation of existence of diabetogenic or anti-diabetogenic effects for VD3 supplementations.

Keywords : VDD, diabetes, T2DM, vitamin D, leptin, obesity, insulin resistance

The combination of omega−3 fatty acids with high doses of vitamin D3 elevate A1c levels: A randomized Clinical Trial in people with vitamin D deficiency

Authors : Abeer Barham, Beisan Mohammad, Luai Hasoun, Shady Awwad, Ibrahim Mosleh, Ahmad Aljaberi, Mahmoud Abu-Samak

Abstract : Purpose: This randomised clinical trial (RCT) was created to assess the influence of 1,25-dihydroxyvitamin D (VD3 ), omega-3 fatty acids (n-3FA) and their combination (D+) on glycated haemoglobin (A1c) levels in Jordanian peoples with vitamin D deficiency (VDD). Participants and methods: This RCT was designed to examine the follow-up (2 months) effect of either 50 000 IU VD3 , 300 mg n-3FA, or the combination of the two supplements on glycated Haemoglobin (A1c) levels in 146 Jordanian women and men with VDD, aged from 25 to 55 years. The eligible participants were randomised into four groups: Control (C); VD3 supplementation (50 000 IU of VD3 was taken weekly) (D3 ); n-3FA supplementation (300 mg of omega-3FA was taken daily) (n-3FA); VD3 and omega-3 supplementation group (D+) with the same protocol as the previous two groups. Results: The combination therapy (n-3FA plus VD3) for 8 weeks significantly increased A1c levels (5.79 ± 0.34 vs 5.41 ± 0.33, P < .001). Tukey test for post hoc comparisons of A1c at follow-up showed that the A1c mean levels were remarkably higher in the D+ study group comparing to the control group (5.78 vs 5.38). Conclusion: The intervention of n-3FA alone or in combination with high doses of VD3 may lead to negative effects on glycaemic control or accelerate the insulin resistance's development in susceptible people for diabetes mellitus (type 2).

Keywords :

The supplementary effects of omega-3 fatty acid alone and in a combination with vitamin D3 on serum leptin levels: A randomized clinical trial on men and women with vitamin D deficiency

Authors : Mahmoud S. Abu-Samak, Luai Z. Hasoun, Abeer Barham, Beisan A. Mohammad, Ibrahim Mosleh, Ahmad Aljaberi, Shady H. Awwad

Abstract : Purpose: This randomized clinical trial (RCT) was designed to assess the effect of VD3, n-3FA, and their combination on serum leptin levels in people with vitamin D deficiency (VDD). Subjects and methods: One hundred and forty six participants, were randomly assigned into four groups supplemented with the dose of 50,000 IU VD3 taken weekly (D), 300 mg n-3FA taken daily (Om), and their combination (D+Om) or control (C) for eight weeks. Fasting baseline and follow-up (10 weeks; 8 weeks supplementation plus washout period of 2 weeks) of serum 25 hydroxyvitamin D (25OHD), leptin, glucose, triglycerides (TG), parathyroid hormone (PTH), calcium, and phosphorus were assayed. A paired T-test was used to assess the changes in serum leptin levels over of the follow-up period. Results: Significant increase in follow-up serum leptin (10.62 ± 7.18 to 14.42 ± 8.29 ng/mL, P = 0.002) and TG (154 ± 84.4 to 200.1 ± 79, P = 0.015) levels were observed in n-3-FA supplemented group. Combination therapy (VD3 plus n-3 FA) significantly increased serum 25OHD (13.49 ± 4.64 to 37.09 ± 11.13 ng/mL, P < 0.001), TG levels (114.3 ± 57.3 to 139.1 ± 60.7 mg/mL, P = 0.007) and insignificantly serum leptin (6.74 ± 4.87 to 8.01 ± 6.77 ng/mL, P = 0.269). Conclusion: Our study referred that notable elevation in leptin and TG levels might be linked to leptin resistance. However, further RCTs are required to clarify possible consequences resulted from the extensive administration of n-3FA supplements and their combinations with high doses of VD3 supplements on humans’ health.

Keywords : Leptin, n-3FA, omega, vitamin D3, diabetes

Dielectric Studies of Tetraaryl and Triaryl Polycarbonates and Comparisons with Bisphenol A-Polycarbonate

Authors : J. J. Fontanella, D. A. Boyles, T. S. Filipova, S. Awwad, C. A. Edmondson, J. T. Bendler, M. C. Wintersgill, J. F. Lomax, M. J. Schroeder

Abstract : The relative permittivity, loss, and breakdown strength are reported for a commercial sample of bisphenol A-polycarbonate (comm-BPA-PC) and a purified sample of the same polymer (rp-BPA-PC) as well as for two new polycarbonates having low molecular cross-sectional areas, namely a copolymer of tetraaryl polycarbonate and BPA-PC (TABPA-BPA-PC) and a triaryl polycarbonate homopolymer (TriBPA-PC). The glass transition temperatures of the new polymers are higher than the Tg of BPA-PC (187 and 191 °C vs. 148 °C). Relative permittivity and loss measurements were carried out from 10 to 105 Hz over a wide temperature range, and results for the α- and γ-relaxation regions are discussed in detail. For the α-relaxation, the isochronal peak position, Tα, scales approximately with Tg. On the other hand, the peak temperature for the γ-relaxation is approximately constant, independent of Tg. Also, in contrast to what is observed for α, γ exhibits a strong increase in peak height as temperature/frequency increases and a significant difference is found between Arrhenius plots determined from isochronal and isothermal data analyses. Next, the γ-relaxation region for comm-BPA-PC and associated activation parameters show strong history/purity effects. The activation parameters also depend on the method of data analysis. The results shed light on discrepancies that exist in the literature for BPA-PC. The shapes of the γ loss peaks and hence glassy-state motions for all the polymers are very similar. However, the intensities of the TriBPA-PC and TABPA-BPA-PC γ peaks are reduced by an amount that closely matches the reduced volume fraction of carbonate units in the two new polymers. Finally, for comm-BPA-PC, the breakdown strength is strongly affected by sample history and this is assumed to be related to volatile components in the material. It is found that the breakdown strengths for TriBPA-PC and TABPA-BPA-PC are relatively close to that for rp-BPA-PC with the value for TriBPA-PC being slightly larger than that for rp-BPA-PC or the value usually reported for typical capacitor grade polycarbonate. Finally, it is shown that the real part of the relative permittivity remains relatively constant from low temperatures to Tg. Consequently, based on the dielectric properties, TriBPA-PC and TABPA-BPA-PC should be usable in capacitors to at least 50 °C higher than BPA-PC.

Keywords : Dielectric properties; dielectric relaxation; glass transition; glassy polymers; high-performance polymers; high temperature materials; polycarbonates

Salivary Cortisol, Testosterone and DHEA in Healthy University Male Students with Hyperleptinemia: Retrospective Cohort Study

Authors : Mahmoud Abu-Samak, Luai Hasoun, May Ibrahim Abu-Taha, Shady Helmi Awwad, Beisan Ali Mohammad

Abstract : Background: Cortisol, testosterone and DHEA are steroid hormones associated with youth physiology. The aim of this study was to evaluate the association of stress hormones with leptin, the hormone of obesity, in university male students. In this prospective cohort study, ninety university male students in the age range of 18 to 24 years were categorized into two groups; NL=Normal or Borderline Serum Leptin Level (<9.4 ng/ml); HL=High Serum Leptin Level (≥ 9.4 ng/ ml). In these groups, serum leptin and salivary levels for testosterone, dehydroepiendisterone (DHEA) and cortisol were immunoassayed. Correlation coefficients model analysis found that M-DHEA was significantly positively correlated with serum leptin in all subjects regardless of study group (r=- 0.248, p=0.003). The model analysis also found that body weight showed greater significant and positive correlation with serum leptin level than did BMI (NL: r=0.549 vs. 0.429, HL; r=0.517 vs. 0.422, respectively). A positive correlation between hyperlipidemia and hyperleptinemia was noted. Negative correlation was seen between M-DHEA and some obesity parameter including BMI and body weight and serum leptin. Elevated cortisol with declined DHEA balance was noted in this study. Our findings shed some light on the potential mechanisms linking obesity, stress and aging hormones in young men where leptin is the main obesity marker that promotes steroid hormones imbalance leading to maladaptation to chronic stress in young men. We observed that serum leptin levels are proportional to DHEA/cortisol imbalance and suggested to be a new indicator of chronic stress maladaptation in young obese males.

Keywords : Salivary hormone; Testosterone; DHEA; Cortisol; Leptin; Obesity; Youth

Associations Between Sleep Deprivation and Salivary Testosterone Levels in Male University Students: A Prospective Cohort Study

Authors : Mahmoud Suleiman Abu-Samak, Beisan Ali Mohammad, May Ibrahim Abu-Taha, Luai Zidan Hasoun, Shady Helmi Awwad

Abstract : Sleep deprivation is a common health problem that is growing rapidly worldwide and it is associated with short- and long-term impacts on health. The aim of this study was to detect potential predictors of salivary testosterone (sT) association with sleep deprivation in Arab male university students. In this prospective cohort study, 77 university male students in the age range of 18 to 26 years were divided into two groups, sleep-deprived (SD) participants and non-sleep-deprived (NSD) participants. Sleep deprivation was defined as sleeping less than 5 hr per night. Blood samples and sT were collected from fasting participants to measure serum levels of glucose, lipid profile, leptin, serotonin, sT, and body mass index (BMI) values. The multiple linear correlation model of high-density lipoprotein cholesterol (HDL-C), BMI, and serotonin was positively correlated with sT ( r = .977, p < .05) in the SD group. No correlations were identified with sT in the NSD group. In the SD study group, the multiple linear regression model of HDL-C, BMI, and serotonin was significantly influenced by sT ( R² = .955, p < .05). These predictors together explained approximately 96% of the variance in sT levels in the SD study group. No predictive variables for sT were reported in the NSD group. Results indirectly confirmed the presence of a positive association between sT and sleep deprivation in young men. This association is mediated by three factors, HDL-C, BMI, and serum serotonin, which are collectively considered as part of a significant physiological adaptation to sleep deprivation in young men.

Keywords : BMI; leptin; lipid; salivary testosterone; serotonin; sleep deprivation

Monitoring of Polychlorinated Biphenyls in Surface Water Using Liquid Extraction, GC/MS, and GC/ECD

Authors : M. Zaater, Y. Tahboub, S. Qasrawy

Abstract : A rapid and reliable analytical method, at trace level concentration was developed and validated for monitoring polychlorinated biphenyls (PCBs) in Jordanian surface water. The method combines the advantage of liquid extraction together with gas chromatography‐mass spectrometry (GC/MS) and gas chromatography‐electron capture detector (GC/ECD). The performance of the method was evaluated by analyzing certified reference material (CRM) of the analytes and applied on real water samples collected from different sites in Jordan. A mixture of 60∶40 dichloromethan‐petroleum ether was chosen as a convenient binary solvent for liquid–liquid extraction. The GC conditions for GC/MS were optimized using He as a carrier gas, temperature programming, and chlorpropham as an internal standard (IS). The conditions for GC/ECD were performed using N2 gas and a temperature program from 160 to 280°C with different increasing rates. The method of GC/MS in the selective ion mode (SIM) gave linear relationships for all PCBs tested between 0.60–6.0 µg/l with R 2=0.9934 (n=7×18). Recoveries from spiked water samples ranged between 87.6 and 91.4%. The mean accuracy and precision obtained were 4.9% and 2.16%, respectively. The mean of detection limit was 0.14±0.04 µg/l. In GC/ECD, linear relationships for all PCBs examined over the range of 0.3–2.4 µg/l was verified as characterized by a linear regression equation and correlation coefficient, R 2=0.9915 (n=12). The average precision and accuracy were 4.86% and 5.21%, respectively. Analyses results clarified that none of the examined Jordanian water samples contained any of the searched for PCBs within the detection limit achieved.

Keywords : Polychlorinated biphenyls, GC/MS, GC/ECD, liquid extraction, monitoring, surface water

Phytochemical analysis and evaluation of the anti-hyperlipidaemic effect of ethanolic leaf extract of Equisetum ramosissimum L. on high-fat diet fed rats.

Authors : Mustafa Al-Bayati, Reem Issa, Mahmoud Abu-Samak, Lilian Alnsour, Shady Awwad.

Abstract :This study aims to investigate the anti-hyperlipidaemic effect of ethanolic leaf extract of Equisetum ramosissimum. 2,2-Diphenyl-1- picrylhydrazyl assay for antioxidants, Folin-Ciocalteu, AlCl3, and UHPLC-MS/MS analysis, focusing on phenols and flavonoid content were performed. Anti-hyperlipidemic effect of the extract on lipid profile and body weight was evaluated alone or in combination with Atorvastatin in rats. The extract was shown to contain phenols (0.032±0.001 μg/g, equivalent to gallic acid), flavonoids (0.044±0.003 mg/g, equivalent to quercetin), and antioxidant IC50 value of (1000.00±0.78 μg/mL). UHPLC-MS/MS analysis revealed the presence of 8 different phenols and flavonoids. An in vivo study on healthy standard diet-fed animals and an induced hyperlipidaemic model showed a significant (P < 0.05) reducing effect of the extract alone and in combination with Atorvastatin on serum lipid profile. These findings revealed the potential advantages of the extract alone and in combination with statins for preventing or treating hyperlipidaemia but need to be further explored.

Keywords : Phenols, flavonoids, anti-hyperlipidemic, Equisetum ramosissimum, high-fat fed rats.

Cytotoxicity Activity of Graviola Fruit Extract with Carbamazepine and Valproic Acid Show Antagonistic and Indifferent Effects.

Authors : Abu Soukhon AA, Abu-Qatouseh L, Mansoor K, El-Hajji FD, Al-Najjar M, Awwad S, Awad R, Omari KW, Mallah E.

Abstract :Objective: Graviola is a tropical fruit with medicinal properties, used for treating various diseases such as inflammation, diabetes, and cancer. Histone deacetylase inhibitors (HDACIs), including carbamazepine (CBZ) and valproic acid (VPA), have been proven strong inhibitors against cancer cell growth. This study investigated the effect of Graviola fruit extract (GFE) on CBZ in healthy rat plasma using high-performance liquid chromatography (HPLC). In addition, the effect of GFE in combination with CBZ and VPA on two human cancer cell lines (PC3 and MCF-7) was explored. Methods: The CBZ levels were analyzed using a simple validated HPLC method. The linearity was achieved at a 0.9998 coefficient of determination over a range of 75–5000 ng/mL CBZ. The MTT assay was used to quantify the percentage of viable cells. Result: The maximum plasma concentration (Cmax) and area under the curve (AUC) for CBZ alone were 4,631 ng/mL and 49,225 ng. h/mL, respectively. However, in the presence of GFE, the values reduced significantly to 2,994 ng/mL and 26,587 ng. h/mL, while the p- value was <0.05. The 3-(4,5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide (MTT) assay results for VPA showed a weak cytotoxicity activity on PC3 and MCF-7 cell lines. Conclusion: A simple and validated HPLC method was used to determine CBZ levels in rats’ plasma. The plasma CBZ levels (Cmax) were significantly reduced in the presence of GFE, indicating the importance of drug-herb interactions. For in vitro studies, two human cancer cell lines, MCF-7 (breast cancer cells) and PC3 (prostate cancer cells), were used to screen the cytotoxicity activity of GFE, CBZ, and VPA. We observed an antagonism effect for GFE and CBZ combination in both cell lines with FIC values > 4. On the contrary, the combination of GFE and VPA showed an additive or indifferent effec

Keywords :Cell lines; cancer; pharmacokinetics.

The discovery of potential tumor necrosis factor alpha converting enzyme inhibitors via implementation of K Nearest Neighbor QSAR analysis.

Authors : Habash, M.; Alshakhshir, S.; Awwad, S.; Abu-Samak, M.

Abstract : Tumor necrosis factor-alpha converting enzyme (TACE) is considered as a pro-inflammatory cytokine which catalyzes the formation of TNF-α from membrane bound TNF-α precursor protein. It has important role in various pathological diseases such as inflammation, anorexia, rheumatoid arthritis, cancer and viral replication. Despite the reporting of a variety of TACE inhibitors of diverse chemical scaffolds whether peptide, peptide-like compounds or others, the bioavailability and pharmacokinetic problems are reflected strongly on its clinical effectiveness. In this effort we employed a novel combination of k-nearest neighbor QSAR modeling to select the best critical ligand-TACE contacts capable of elucidation of TACE inhibitory bioactivity among a long list of inhibitors. The recurrence of one valid pharmacophore hypothesis among the successful models emerged the pharmacophore to be used as 3D search queries to screen the National Cancer Institute’s structural database for new TACE inhibitors. Eight potent TACE inhibitors were discovered with inhibition percentage exceeding 50% at 100 μM inhibitor concentration.

Keywords : Tumor necrosis alpha converting enzyme (TACE), k nearest neighbor, Ligandfit, Pharmacophore, QSAR, In vitro assay.

Combined effect of omega-3 fatty acid and vitamin D3 on oxidized LDL-C and non-HDL-C levels in people with vitamin D deficiency: A randomized controlled trial.

Authors : Mehdawi, Amani; Mohammad, Beisan; Mosleh, Ibrahim; Khader, Heba A; Habbash, Maha; Nassar, Razan; Awwad, Shady; Hasoun, Luai; Abu-Samak, Mahmoud.

Abstract :The present randomized clinical trial (RCT) was conducted on Jordanian participants with vitamin D deficiency (VDD) with no other medical conditions, to evaluate the combined effect of 1,25-dihydroxy vitamin D 3 (Vit.D 3 ) and omega-3 fatty acid (n-3FA) supplements (D+) on oxidized low-density lipoprotein (Ox-LDL) and non-high-density lipoprotein cholesterol (non-HDL-C) levels as common predictors of cardiovascular diseases (CVDs). Participants were randomized into 4 groups as follows: a control group (C) that received no supplementations, a Vit.D 3 group that received 50,000 IU of Vit.D 3 every week, an n-3FA group that received 300 mg of omega-3 fatty acid every day, and a D+ group that received a combination of both supplements, with the same dosage administered by the previous groups but with a 4-6-hour time interval between Vit.D 3 and n-3FA administration to avoid any possible interaction. All supplementations were administered orally for 8 weeks. Forty-seven participants were allocated to each group. Twenty-six in the control group, 37 participants in the Vit.D 3 group, 37 participants in the n-3FA group, and 46 participants in the D+ group completed the study to the end. The D+ supplementations significantly increased non-HDL-C (118.99 ± 60.98 to 155.26 ± 43.36 mg/dL, P << 0.05) but decreased Ox-LDL-C levels (69.29 ± 37.69 to 52.81 ± 17.30 pg/mL, P = 0.03). The stepwise regression showed that the serum LDL-C level was the main independent variable involved in the elevation of non-HDL levels (R 2 = 0.837) observed at the end of the trial in the D+ group. The groups that were supplemented with either Vit.D 3 alone or n-3FA alone had an insignificant decrease in the level of Ox-LDL-C. In conclusion, despite the observed hyperlipidemic effect, the combination treatment is recommended by the research team because the decrease in Ox-LDL may offset the hyperlipidemic effect.

Keywords :vitamin D deficiency, vitamin D3, omega-3 fatty acids, non-HDL, oxidized-LDL cholesterol, LDL-C, TG.

The effect of excessive coffee consumption, in relation to diterpenes levels of medium-roasted coffee, on non–high-density lipoprotein cholesterol level in healthy men

Authors :Awwad, S.; Abu-Zaiton, A; Issa, R.; Said, R.; Sundookah, A.; Habash, M.; Mohammad, B.; Abu-Samak, M.

Abstract :This study was designed to determine the levels of coffee oil and diterpenes and evaluate the correlation between the effect of excessive roasted coffee consumption on non-high-density lipoprotein cholesterol (non-HDL) and the roasting degree effect on the levels of coffee oil and diterpenes extracted from Coffea arabica. The coffee oil and diterpenes were extracted using soxhlet and liquid-liquid extraction. Sixty-six healthy normolipidemic male participants were assigned for this study which consisted of two stages. The first stage is the coffee abstaining stage where subjects were requested to abstain from drinking coffee for 2 weeks. The second stage is the coffee drinking stage which consisted of groups (the control group and coffee-drinking group). The levels of TC, TG, LDL-C, HDL-C, and non-HDL were determined in all participants before and after the experiment. The results indicated that the coffee roasting degree demonstrated a significant increase in the levels of coffee oil and diterpenes ranging from 9.31% (green coffee) to 15.64% (dark roast) and from 0.205% (green coffee) to 0.300% (dark roast) for diterpenes. In conclusion, the current study revealed that excessive consumption of medium roasted coffee was associated with elevated non-HDL levels in normotensive nonsmoker healthy men which might be attributed to the positive association between the degree of roasting and diterpenes levels.

Keywords : Coffee, non-HDL, lipid profile, coffee oil, diterpenes, roasting degree

Research Title :Antioxidant Activity and UHPLC-MS/MS Characterization of Polyphenol and Nicotine Content in Nicotiana Glauca Leaf Extracts: A Comparative Study of Conventional and Deep Eutectic Solvent Extraction Methods

Authors : Issa R., Al-Akayleh F., Alnsour L., Al-Sammarraie T.R., Omari K.W., Awwad S.H.

Abstract :The leaves of Nicotiana glauca (N. glauca; Solanaceae) plant are a known, major human health concern. This study investigated the antioxidant activity and polyphenols composition of aerial parts of N. glauca collected from its wild habitat in Jordan, using Methanol-Conventional (MC) and deep eutectic solvents (DES) extraction methods in addition to nicotine content determination using UHPLC. Our results showed that the MC extract contains fewer total phenols and flavonoid content than the 90% DES extract, (0.1194 ± 0.009 and 0.311 ± 0.020 mg/mL equivalent to gallic acid) and (0.01084 ± 0.005 and 0.928 ± 0.09 mg/mL equivalent to rutin), respectively. Moreover, this study showed that the prepared MC extract contain 635.07 ppm nicotine, while the 90% DES extract contain 1194.91 ppm nicotine. Extracts prepared using the MC and the DES methods exhibited weak antioxidant activities; the highest was a 33% inhibition rate (equivalent to ascorbic acid), obtained by the 90% DES extract,. The performed UHPLC-MS/MS analysis in this study also revealed the presence of variations in the detected compounds between the two extraction methods. Furthermore, this study found that environmentally friendly DES extraction of N. glauca produced higher phenol and flavonoid content than the MC method; this highlights the superior efficiency and environmental benefits of sustainable chemistry methods for extracting valuable phytoconstituents.

Keywords :Nicotiana glauca; chromatography; mass spectrometry; antioxidants; deep eutectic solvents; sustainable extraction

Research Title :The effect of roasting degrees on bioactive compounds levels in Coffea Arabica and their associations with glycated hemoglobin levels and kidney function in diabetic rats

Authors : Al-Tamimi O., Awwad S.H., Issa R, Al-Qaisi T., Abazid H., Daraosheh A., Mohammad B., Filipova T., Abu-Samak M.

Abstract :The current study aims to assess the content of coffee oil, diterpenes, caffeine, and chlorogenic acid in coffee beans, and to evaluate the effects of roasting degrees on bioactive compound levels in Coffea arabica and their associations with glycated hemoglobin levels and kidney function parameters in streptozotocin-induced diabetic rats. These constituents were quantified using Soxhlet, liquid–liquid extraction (LLE), and high-performance liquid chromatography (HPLC)-diode array detecto (DAD). The outcomes exhibited a positive correlation between levels of coffee oil and diterpenes with a roasting degree. However, it demonstrated a negative correlation between caffeine and chlorogenic acid with a roasting degree. The male rats were categorized into two sections: healthy (nondiabetic) and streptozotocin-induced diabetic groups. Rats have daily administrated 2 ml of coffee extract using an oral gavage subjected to different roasting degrees for 3 weeks. Fasting blood glucose (FBG), urea, and creatinine in serum were assayed at baseline and end of the experiment. The highest significant elevation (Pa = 0.016) in the mean urea and creatinine levels was noted in the diabetic group administrated with dark coffee extract whereas the lowest elevation was in the diabetic group administrated with green coffee extract. In conclusion, our study has shown that coffee roasting degree may have a potential negative effect on kidney function that may accelerate or exacerbate diabetic nephropathy in induced diabetic rats. A noteworthy change in urea levels was observed in diabetic rats treated with dark-roasted coffee, which might be attributed to the decline in antioxidant constituents: chlorogenic acids and diterpenes.

Keywords : Nephropathy, coffee, quantification, diabetes, caffeine, chlorogenic acids

Research Title :Nephroprotective effects of Equisetum ramosissimum L. extract in streptozotocin-induced diabetic rats.

Authors :Abdullah R.K., Issa R.A., Abu-Samak M., Mohammad B.A., Abbas M.A., Awwad S.H.

Abstract :Diabetes is a widespread health issue that impacts people all over the globe. The Equisetum ramosissimum L. plant has numerous traditional uses and pharmacological properties, including antidiabetic effects. The objective of this study was to thoroughly examine the advantages of incorporating extracts from the aerial components of E. ramosissimum to control diabetic nephropathy. The phytochemical constituents of E. ramosissimum extract were explored using phytochemical and HPLC analysis, focusing on phenols and flavonoid content. The effect of plant extract was evaluated on different kidney function parameters linked to diabetic nephropathy (fasting blood glucose, creatinine, uric acid, and urea) in streptozotocin induced-diabetic rats. Histopathological changes in the liver were also examined. The results showed methanol and ethanol extracts of E. ramosissimum have a total content of phenols (equivalent to gallic acid, 7.62 and 8.97 mg/g) and flavonoids (equivalent to quercetin, 8.87 and 12.86 mg/g), respectively. After conducting the UHPLC-MS/MS analysis, it was found that both the methanol and ethanol extracts contained isoferulic acid, ISO-Orientin, myristic acid, linoelaidic acid, rutin, and 3-Glu-7-Rha quercetin. Additionally, isoferulic acid, myristic acid, linoelaidic acid, rutin, and 3-Glu- 7-Rha quercetin were present in the extracts. The ethanol extract of E. ramosissimum significantly impacted STZ-induced diabetic mice by reducing their fasting blood glucose levels, and their creatinine and urea levels (P < 0.005). In conclusion, E. ramosissimum ethanol extract has shown potential effects to counteract some diabetes consequences on kidney function. Therefore, further studies are required to investigate its effect on other diabetes-related complications.

Keywords : Equisetum ramosissimum, diabetes, nephroprotective, kidney functions, phenols, flavonoids.

Research Title :Correlation of High Serum Leptin Levels with selected stress-associated salivary hormones in Arab Healthy Young Men

Authors :Abu-Samak, M., Mohammad, B. A., Awwad, S. H., Barham, A., Abu-Zaiton, A.

Abstract :Abstract: Background: Allostatic load is accompanying the youth’s modern lifestyle. The aim of this study is to evaluate the association of stress markers such as salivary cortisol and dehydroepiandrosterone (DHEA) hormone levels in Arab young men with high levels of leptin. Methods: 118 males aged 18–26 years, were eligible to participate in a retrospective cross-sectional study. Participants were categorized into two groups: moderate levels of serum leptin (<7 ng/ml) and high levels of leptin levels (≥7 ng/ml). Serum leptin, salivary cortisol and DHEA, lipid profile, and fasting blood glucose were assayed for each participant. Results: A significant decrease in the mean levels of morning salivary- DHEA was observed in high levels of leptin compared to moderate levels of serum leptin study group (p<0.0001). A significant increase was also observed in mean values of body weight and body-mass index. A positive correlation was observed between morning salivary- DHEA and serum-leptin levels in all subjects (p=0.003). Pearson correlation showed significant correlations between morning salivary- DHEA, body weight, body mass index, and triglyceride. Conclusion: The serum leptin level was proportional to the DHEA/cortisol imbalance that was associated with allostatic load and the inability to accommodate stress in male youth.

Keywords :Allostatic load BMI Cortisol DHEA Leptin obesity Youth

Research Title :A comparison of the effect of omega-3 alone versus omega-3/ Vitamin D3 co-supplementation therapy on 25-hydroxyvitamin D levels in adults with vitamin D deficiency.

Authors :Beisan Mohammad, Ghaleb Oriquat, Heba Khader, Husni Farah, Ahmad Alsayed, Jehad Alhmoud, Mohammad Al-Najjar, Luai Hasoun, Shaimaa Abdelhalim, Shady Awwad, Mahmoud Abu-Samak.

Abstract :Background: Omega-3 fatty acid (n-3FA) supplementation may improve total hydroxyvitamin D (25OHD) levels in people with normal or low vitamin D levels. However, there was no agreement on the potential therapeutic benefit of n-3FA on vitamin D toxicity. Objective: The current randomized controlled clinical study (RCT) sought to determine if daily omega-3 fatty acid (n-3FA) supple-mentation affected 25-hydroxyvitamin D (25OHD) levels. Methods: One hundred and twenty Jordanians aged 22 to 55 were randomly allocated into three groups supplemented with the dose of 50,000 IU VD3 taken weekly (D3) alone, 300 mg n-3FA taken daily (n-3FA) alone, or their com-bination (50,000 IU VD3 taken weekly + 300 mg n-3FA taken daily (D+). In addition, fasting baseline and follow-up (ten weeks; eight weeks supplementation plus two weeks washout) of serum 25 hydroxyvitamin D (25OHD), parathyroid hormone (PTH), calcium, and leptin were assayed. Results: The n-3FA supplementation significantly decreased mean serum 25OHD levels (11.97±4.6 vs 16.5±4.8, P<0.001) compared with their peers in the D3 group who showed a significant increase in 25OHD levels (41.15±11.7 vs 19.34±6.8, P<0.001). The follow-up PTH mean levels were pro-portionally accompanied by significant alterations of 25OHD levels in the two study groups. Conclusions: This novel RCT provides that eight weeks of n-3FA therapy significantly reduced 25OHD levels, which adverse effects may accompany vitamin D status and its health-related consequences, such as bone homeostasis, cardiovascular disease, and COVID-19 conditions.

Keywords : Vitamin d3; omega-3; vitamin d toxicity; cardiovascular disease; covid-19.

Research Title :Development and Validation of a Novel Stability-Indicating RP-HPLC Method for The Simultaneous Quantitation of Amlodipine, Glimepiride, and Perindopril Arginine.

Authors : Shady Awwad, Samar Thiab, Reem Issa, Hassan Abul-Futouh, Laith MH Al-Shdifat, Ahmad Aljaberi, Asaad Shahin, Dima Albals, Beisan Mohammad, Mahmoud Abu-Samak.

Abstract :Abstract: An accurate, precise, simple and stability-indicating RP-HPLC method was developed and validated for the simultaneous quantitation of perindopril arginine (PA), amlodipine (AML) and glimepiride (GLM) in solution in accordance with the International Conference on Harmonisation (ICH) guidelines. The HPLC analysis was performed using a Supelcosil LC-18 column (25 cm × 4 mm × 5 μm), at a flow rate of 1.0 mL/min, with wavelength of detection at 220.0 nm for PA, AML and GLM. The optimised mobile phase consisted of a mixture of acetonitrile, methanol and phosphate buffer in the ratio of (20:50:30) (v/v/v) which was adjusted to pH 3.5 ± 0.1 with o-phosphoric acid. The proposed method was validated for linearity, system suitability, accuracy, precision (intra- and inter-day) and robustness. The HPLC results displayed good separation and resolution in a total analysis time of 16 minutes where PA, AML and GLM were eluted at retention times of 4.408 min, 5.415 min and 12.189 min, respectively. The optimised method displayed good linearity over the concentration range of 0.25 - 2.00 mg/mL for PA and 0.10 - 0.75 μg/mL for AML and GLM. The recoveries ranged from 98.0 to 102.0% and the RSD% values were found to be less than 2.00 for all drugs. The combination of three drugs was exposed to several forced degradation studies such as acid, base, neutral, thermal, oxidative and photolytic conditions. The forced degradation studies indicated that the optimised method is valid for stability studies.

Keywords :Keywords: Amlodipine, glimepiride, perindopril arginine, HPLC, validation, stability.

Research Title :Quantification of Bioactive Compounds in Coffea Arabica and Their Impact with Omega-3 Supplementation on Non-High-Density Lipoprotein Levels in Hyperlipidemia-Induced Rats.

Authors :Sojoud M. Al-Zoubi, Shady H. Awwad, Reem Issa, Ahmad Daraosheh, Talal Al-Qaisi, Husni Farah, Khaled Omari, Beisan A. Mohammad, Mumen Amer, Rula F. Khuzaie, Mahmoud S. Abu-Samak.

Abstract :This study aims to evaluate the levels of coffee lipids, diterpenes, chlorogenic acid (CGA), and caffeine in coffee samples with different roasting levels. Additionally, it aims to assess the impact of coffee extracts with/without omega-3 supplementation on lipid profile parameters. Liquid-liquid and soxhlet extraction, and quantitation were performed using HPLC-DAD. Two groups of Wistar rats were used: non-hyperlipidemic and hyperlipidemic. Blood samples were collected before and after hyperlipidemia induction after six weeks. The lipid profile parameters including total cholesterol (TC), low-density lipoprotein (LDL), high-density lipoprotein (HDL), triglyceride (TG), non-high-density lipoprotein (non-HDL), and total cholesterol-to-high-density lipoprotein ratio (TC/HDL) were determined. A positive correlation between coffee lipids, diterpenes, and caffeine levels with the coffee’s roasting degree was found, and a negative correlation for CGA. The study was conducted on rats fed with a healthy and hyperlipidemia-induced diet. The green coffee had a significant effect on the lipid profile of non-hyperlipidemic rats. It lowered the levels of TG, LDL, and TC/HDL, and raised HDL and non-HDL levels significantly compared to the control group. In combination, green coffee and omega-3 lowered TC, TG, LDL, non-HDL, and TC/HDL levels. Green coffee alone or in combination with (omega-3) lowered lipid profile parameters but increased non-HDL levels in non- and hyperlipidemic induced-rats.

Keywords:Caffeine, chlorogenic acid, Coffee, diterpenes, lipid profile, omega-3

Research Title :Effect of Eicosapentaenoic Acid, Docosahexaenoic Acid and Their Combination on Selected Atherogenic Biomarkers in High-Fat Diet Rat Model.

Authors :Abu Sadah AH, Issa R, Farah H, Oriquat G, Awwad SH, Mosleh I, Mohammad B, Aljaberi A, Al-Najjar M, Abu-Samak MS.

Abstract :Findings concerning eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA) as well as omega-3 (n-3FA) effects on lipid profile and biomarkers of atherosclerosis progression are still highly debated. The current study was designed to evaluate and compare the effect of EPA, DHA, and their combination in the form of n-3FA on serum non-high-density lipoprotein (NHC), oxidized-low-density lipoprotein (Ox-LDL), Lipoprotein(a) Lp(a), and other lipid profile parameters levels in rats with high-fat diet model. Based on the diet and supplementation model, six groups (n = 6 per group) of male Westar rats were distributed as follows: standard diet (SD), high-fat diet (FD); FD + atorvastatin (ATV), FD + omega-3 (n-3FA), FD + EPA (EPA), and FD + DHA (DHA). The results have shown a significant higher mean NHC levels in the DHA and n-3FA groups than in the EPA group (27.52 ± 2.92 vs. 43.23 ± 8.98 and 45.65 ± 5.08 mg/ dl, respectively, p < 0.001). In addition, the mean levels of total cholesterol and Lp(a) levels were significantly higher in DHA than in EPA (35.8 ± 2.04 vs. 51.8 ± 6.33 mg/dl, 2.42 ± 0.71 vs. 4.41 ± 1.14 ng/dl, p < 0.001). Significant higher mean Ox-LDL levels were observed in n-3FA than in DHA (p < 0.001) or EPA (p < 0.05). No significant in mean Ox-LDL levels was observed between EPA and DHA study groups (t = 3.62, p = 0.1387). The current study findings revealed the potential advantages of EPA supplements but not DHA supplements alone or their combination with EPA in the common form known as omega-3 for preventing or treating hyperlipidemia.

Keywords :Omega-3, DHA, EPA, Ox-LDL, non-HDL, lipid profile, lipidemia.

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